From 99f151b43f13963bfc84c6c0bebb2c4c0e713706 Mon Sep 17 00:00:00 2001 From: laurabiggins Date: Thu, 10 Aug 2023 15:11:17 +0100 Subject: [PATCH 01/15] Added new tab in for aging t-SNE plots, which currently show 2 points. --- expressionViewer/app.R | 5958 ++++++++++++++++++++-------------------- 1 file changed, 3022 insertions(+), 2936 deletions(-) diff --git a/expressionViewer/app.R b/expressionViewer/app.R index 30a766b..f6d5e0c 100644 --- a/expressionViewer/app.R +++ b/expressionViewer/app.R @@ -1,2936 +1,3022 @@ - - - -#importing libraries & functions, reading in data, and setting up global options -{ - #setting global options - options(scipen = 5, digits = 4) - - #libraries - { - library(ggExtra) - library(shiny) - library(DESeq2) - library(shinyjs) - #library(shinyBS) - library(DT) - library(ggplot2) - library(ggrepel) - library(viridis) - library(plotly) - library(stringr) - library(htmlwidgets) - library(shinyWidgets) - library(shinycssloaders) - library(png) - library(htmltools) - #library(shinythemes) - #library(shinydashboard) - #library(dashboardthemes) - library(rintrojs) - library(shinyalert) - - } - - # #functions and scripts - # source("customFunctions.R") - - #data - { - #deseq2 object - DESeq_object <- readRDS(file = "data/DESeq_Object.rds") - - #formatting names for available contrasts - results_names <- resultsNames(DESeq_object) - #print(results_names) - - #aesthetic formatting of name labels - { - names(results_names) <- gsub(pattern = "tissue", - replacement = "", - x = results_names, - ignore.case = T) - - names(results_names) <- gsub(pattern = "\\.cellTypeNT", - replacement = "_Tconv", - x = names(results_names), - ignore.case = T) - - names(results_names) <- gsub(pattern = "\\.cellTypeT", - replacement = "_Tregs", - x = names(results_names), - ignore.case = T) - - # names(results_names) <- gsub(pattern = "_NT$", - # replacement = "_CD4", - # x = names(results_names), - # ignore.case = T) - # - # names(results_names) <- gsub(pattern = "_T$", - # replacement = "_Treg", - # x = names(results_names), - # ignore.case = T) - - names(results_names) <- gsub(pattern = "lymphnode", - replacement = "LN", - x = names(results_names), - ignore.case = T) - - names(results_names) <- gsub(pattern = "intraepitheliallayer", - replacement = "IEL", - x = names(results_names), - ignore.case = T) - - names(results_names) <- gsub(pattern = "laminaproprialymphocyte", - replacement = "LPL", - x = names(results_names), - ignore.case = T) - - names(results_names) <- gsub(pattern = "peyerspatches", - replacement = "PP", - x = names(results_names), - ignore.case = T) - - names(results_names) <- gsub(pattern = "_", - replacement = " ", - x = names(results_names), - ignore.case = T) - } - - - #meta data - MGI_info <- read.delim(file = "data/combined_MGI_info.txt", - sep = "\t", - stringsAsFactors = F) - - #counts - mean_norm_counts <- readRDS(file = "data/Mean_Normalised_Counts.rds") - mean_norm_counts$Gene.stable.ID <- gsub(pattern = "\\.[0-9]+$", - replacement = "", - x = rownames(mean_norm_counts), - ignore.case = T) - #mean_norm_counts2 <- readRDS(file = "Mean_Normalised_Counts.rds") - #aesthetic formatting of mean_norm_counts - { - colnames(mean_norm_counts) <- gsub(pattern = "_NT$", - replacement = "_Tconv", - x = colnames(mean_norm_counts), - ignore.case = T) - - colnames(mean_norm_counts) <- gsub(pattern = "_T$", - replacement = "_Treg", - x = colnames(mean_norm_counts), - ignore.case = T) - - colnames(mean_norm_counts) <- gsub(pattern = "lymphnode", - replacement = "LN", - x = colnames(mean_norm_counts), - ignore.case = T) - - colnames(mean_norm_counts) <- gsub(pattern = "intraepitheliallayer", - replacement = "IEL", - x = colnames(mean_norm_counts), - ignore.case = T) - - colnames(mean_norm_counts) <- gsub(pattern = "laminaproprialymphocyte", - replacement = "LPL", - x = colnames(mean_norm_counts), - ignore.case = T) - - colnames(mean_norm_counts) <- gsub(pattern = "peyerspatches", - replacement = "PP", - x = colnames(mean_norm_counts), - ignore.case = T) - } - - norm_counts <- DESeq2::counts(object = DESeq_object, normalized = T) - #aesthetic formatting of norm_counts - { - colnames(norm_counts) <- gsub(pattern = "_NT", - replacement = "_Tconv", - x = colnames(norm_counts), - ignore.case = T) - - colnames(norm_counts) <- gsub(pattern = "_T", - replacement = "_Treg", - x = colnames(norm_counts), - ignore.case = T) - - colnames(norm_counts) <- gsub(pattern = "lymphnode", - replacement = "LN", - x = colnames(norm_counts), - ignore.case = T) - - colnames(norm_counts) <- gsub(pattern = "intraepitheliallayer", - replacement = "IEL", - x = colnames(norm_counts), - ignore.case = T) - - colnames(norm_counts) <- gsub(pattern = "laminaproprialymphocyte", - replacement = "LPL", - x = colnames(norm_counts), - ignore.case = T) - - colnames(norm_counts) <- gsub(pattern = "peyerspatches", - replacement = "PP", - x = colnames(norm_counts), - ignore.case = T) - } - - #PCA, tSNE and UMAP plots - PCA_means_plots <- readRDS(file = "data/PCA_plots_means.rds") - PCA_plots <- readRDS(file = "data/PCA_plots.rds") - tSNE_plots <- readRDS(file = "data/tSNE_plots.rds") - UMAP_plots <- readRDS(file = "data/UMAP_plots.rds") - - #importing min. transcript per cell gene list object - filtered_genes_lists <- readRDS(file = "data/Filtered_Gene_Lists.rds") - - #importing TCR plots - TCR_plots_list <- list.files(path = "www/TCR_plots/", pattern = ".png") - #print(TCR_plots_list) - - #importing Pathview plots - { - Pathview_plots_list <- list.files(path = "www/Pathview_plots", - pattern = ".multi\\.png") - #print(Pathview_plots_list) - - #mapping Pathview plot files to pathway names - Pathview_mapping <- read.delim(file = "data/output_gage.tab", - sep = "\t", stringsAsFactors = F)[-1,1:2] - Pathview_mapping <- - Pathview_mapping[Pathview_mapping[,1] %in% - gsub(pattern = "\\..*", - replacement = "", - x = Pathview_plots_list, - ignore.case = T),] - Pathview_mapping <- Pathview_mapping[order(Pathview_mapping[,1], - decreasing = F),] - - } - - } - - #importing functions - source("R/customFunctions.R") - -} - - -#shiny app ui -{ - # Define UI ---- - ui <- fluidPage( - - # using shinyjs to enable or disable certain sections/elements of the app - shinyjs::useShinyjs(), - # using shinyalert to enable popup messages - shinyalert::useShinyalert(), - - # Step-by-step tutorial - introjsUI(), - - #importing shinydashboard css - # tags$head( - # includeCSS(file.path('www', path = "AdminLTE.css")), - # includeCSS(file.path('www', path = "shinydashboard.css") - # )), - - #shinythemes::themeSelector(), - #theme = shinytheme("cerulean"), - #shinyDashboardThemes(theme = "grey_light"), - #shinyDashboardThemes(theme = "grey_dark"), - - #dashboardBody(), - - # App title ---- - #titlePanel("Expression Viewer"), - titlePanel("The Tissue Tregs Project"), - - tabsetPanel( - - #Landing Page panel - { - tabPanel(title = "The Study", value = "TheStudy", - - h3("General description/information of the study/data"), - - p(paste0("Some description of the study, as well as where to ", - "find additional sources such as papers and data.")), - p(paste0("Ask Adrian and Oliver as they would be able to ", - "provide a much better description")), - - hr(), - - column(width = 9), - - column(width = 3, - # style = paste0('margin-bottom:30px;border:1px solid; ', - # 'padding: 10px;'), - - wellPanel( - h4("Supplementary material for download"), - - h5("Bulk RNA-seq analysis"), - - withTags( - ul( - li(downloadLink( - outputId = "MultiQCBulk", - label = "QC and pre-processing report" - )), - li(downloadLink( - outputId = "GlobalBulk", - label = "PCA, t-SNE and UMAP plots" - )), - li(downloadLink( - outputId = "DiffExprsBulk", - label = paste0("Counts and differential ", - "expression results") - )), - li(downloadLink( - outputId = "GSEABulk", - label = "Gene set enrichment results" - )) - ) - ), - - # downloadLink( - # outputId = "MultiQCBulk", - # label = "QC and pre-processing report" - # ) - - ) - ), - - br(), - br(), - br(), - br(), - br(), - - fluidRow( - h6(paste0("Copyright 2020, Liston Lab, Babraham Institute. ", - "Babraham internal/intranet release only. ", - "Copyright licence will most likely change by the ", - "first public release as per the requirements of ", - "the publication journal and BI.")) - ) - ) - }, - - #Global Analysis panel - { - tabPanel(title = "Global Analysis", value = "GlobalAnalysis", - - h3("Exploratory analysis of the expression data"), - - p(paste0("PCA, t-SNE and UMAP analysis and plots of the ", - "RNA-seq expression data from the study.")), - - p(paste0("Plots can be downloaded by right clicking the ", - "plot and clicking on 'Save image as...'")), - - #br(), - hr(), - - h4("Bulk RNA-seq data"), - - fluidRow( - - column(width = 12, - - selectizeInput(inputId = "TriplePlot", - label = "Select Plot Type", - # choices = as.list(results_names), - #choices = c("", "PCA", "t-SNE", "UMAP"), - choices = c("PCA - tissue means", - "PCA", "t-SNE", "UMAP"), - selected = "PCA - tissue means", - #selectize = T, - multiple = F, - options = list( - placeholder = "Click to select") - ), - ) - ), - - fluidRow( - - column(width = 4, - plotOutput(outputId = "PCAplot1") %>% - withSpinner(type = 3, size = 0.5, - color.background = "#FFFFFF") - ), - column(width = 4, - plotOutput(outputId = "PCAplot2") %>% - withSpinner(type = 3, size = 0.5, - color.background = "#FFFFFF") - ), - column(width = 4, - plotOutput(outputId = "PCAplot3") %>% - withSpinner(type = 3, size = 0.5, - color.background = "#FFFFFF") - ) - ), - - hr(), - - h4("Single cell RNA-seq data"), - - p("Will be available soon") - - ) - }, - - #Bulk RNA-seq Expression panel - { - tabPanel(title = "Bulk RNA-seq Expression", - value = "BulkDiffExprs", - - fluidRow( - - column(width = 7, - h3("Generate contrasts"), - p(paste0("The differential expression analysis is performed ", - "using DESeq2. For a quick walkthrough on doing the ", - "the analysis, please click the yellow button with ", - "the question mark.")) - ), - - column(width = 5, - br(), - br(), - # actionButton(inputId = "TutorialButton", - # label = "Start tutorial"), - actionBttn(inputId = "TutorialButton", - label = "Start tutorial", - style = "material-circle", - color = "warning", - icon = icon("question")), - ) - ), - - hr(), - - fluidRow( - - column(width = 2, - - wellPanel( - - fluidRow( - - column(width = 9, - h4("Available Contrasts"), - ), - - column(width = 3, - - dropdownButton(inputId = "mydropdown", - circle = TRUE, - #status = "info", - status = "secondary", - #icon = icon("question"), - icon = icon("info"), - right = F, up = F, - width = "500px", - tooltip = tooltipOptions( - title = "Instructions"), - - h4("To Begin:"), - - h5(paste0("Start by ", - "selecting tissues", - " for the contrast", - " groups in this ", - "panel. This will", - " enable the ", - "'Generate ", - "Contrasts' button", - " at the bottom. ", - "A tissue cannot ", - "be in both ", - "groups ", - "simultaneously.")), - - h5(paste0("The drop down ", - "list and 'Add' ", - "button below ", - "each of the two ", - "selections can ", - "be used to add ", - "relevant tissues", - ", or to ", - "clear the current", - " selection. The ", - "buttons will not ", - "duplicate ", - "tissues that are ", - "already selected." - )), - - h5(paste0("Clicking the ", - "'Generate ", - "Contrasts' button", - " will perform ", - "the differential ", - "expression ", - "analysis between ", - "the two contrast ", - "groups, creating ", - "a table of ", - "results and a ", - "volcano plot on ", - "right." - )), - - h5(paste0("Clicking the ", - "'Show Advanced Options' ", - "checkbox reveals three ", - "additional options to ", - "adjust the analysis and ", - "visualisation. The ", - "'Minimum Transcripts per ", - "Cell' drop down list ", - "gives options to filter ", - "results to remove low ", - "expression genes if ", - "needed. The 'log2 Fold ", - "Change' and 'Adjusted ", - "p.value' fields are the ", - "significance criteria ", - "used in the volcano plot ", - "and can be adjusted as ", - "needed." - )), - - hr(), - h4("Interactivity:"), - - h5(paste0("The table and ", - "volcano plot are ", - "interactive. Up ", - "to 50 points/rows", - " can be selected ", - "to mark them in ", - "the volcano plot ", - "and to generate ", - "a plot of ", - "normalised counts", - " of the selected ", - "genes across all ", - "samples.")), - - h5(paste0("Alternatively, ", - "points can also", - " be selected by ", - "clicking on them", - " in the volcano", - " plot to mark ", - "them in the plot", - " and also select", - " the respective", - " rows in the ", - "table. Clicking", - " a selected ", - "point again will", - " deselect it in ", - "the plot and ", - "table.")), - - h5(paste0("The selection of ", - "points/rows can ", - "be reset at any ", - "time using the ", - "'Reset Row ", - "Selection' button", - " above the table." - )), - - h5(paste0("The plots can be ", - "zoomed in by ", - "dragging a ", - "selection box ", - "over the relevant", - " area. Reset the ", - "zoom by either ", - "double clicking ", - "anywhere in the ", - "plot or by ", - "clicking the home", - " icon on the top ", - "right of the ", - "plot.")), - - h5(paste0("The table can be ", - "filtered through ", - "the fields under ", - "the column ", - "headers. These ", - "can also be used ", - "to search the ", - "column of ", - "interest.")), - - h5(paste0("Additionaly, the ", - "table can be ", - "sorted by ", - "multiple columns ", - "by shift clicking", - " the column ", - "headers.")), - - hr(), - h4("Download/Export:"), - - h5(paste0("The results table", - " can be ", - "downloaded as ", - "tab delimited ", - "files from the ", - "three download ", - "buttons above the", - " table.")), - - h5(paste0("The plots can be ", - "downloaded via ", - "the camera icon ", - "on the top right ", - "of each plot.", - "Downloading a ", - "plot while zoomed", - " in will download", - " the zoomed in ", - "view.")), - - h5(paste0("The 'Show Plot ", - "Export Options' ", - "that becomes ", - "available above ", - "the volcano plots", - " reveals options ", - "to modify the ", - "export type and ", - "size of the ", - "plot. The ", - "format of the ", - "image (SVG/PNG/", - "JPEG) can be ", - "selected from ", - "the radio button ", - "that becomes ", - "available below ", - "the 'Generate ", - "Conrasts' button.", - " The size of the ", - "plot can be ", - "adjusted by ", - "specifying the ", - "height and width ", - "in pixels." - )) - ) - ), - ), - #h4("Available Contrasts"), - - selectizeInput(inputId = "Group1", - label = "Contrast Group 1", - # choices = as.list(results_names), - choices = list( - Tregs = results_names[ - grepl(pattern = "cellTypeT", - #grepl(pattern = "_Treg", - x = results_names, - ignore.case = T)], - Tconv = results_names[ - grepl(pattern = "cellTypeNT", - #grepl(pattern = "_Tconv", - x = results_names, - ignore.case = T)] - ), - selected = NULL, - #selectize = T, - multiple = T, - options = list( - placeholder = paste0("Click here ", - "to manually", - " select ", - "...") - ) - #) - ), - - #fluidRow( - #h5("Select available:"), - #h5("Quick add available:"), - # actionButton(inputId = "Group1SelectTconv", - # label = "Tconv"), - # actionButton(inputId = "Group1SelectTregs", - # label = "Tregs"), - selectizeInput(inputId = "Group1Selector", - label = "Add available:", - choice = list( - Treg = c("All Tregs", - "Blood Tregs", - "Lymphoid Tregs", - "Non-lymphoid Tregs", - "Gut-associated Tregs"), - Tconv = c("All Tconv", - "Blood Tconv", - "Lymphoid Tconv", - "Non-lymphoid Tconv", - "Gut-associated Tconv") - ), - selected = "All Tregs", - multiple = F, - options = list( - placeholder = "Click to select") - ), - actionButton(inputId = "Group1Add", - label = "Add"), - actionButton(inputId = "Group1Clear", - label = "Clear Selection"), - #), - - #br(), - #br(), - hr(), - #br(), - - #output of group 1 selection for debugging -- remove/ - # hide when deploying - #verbatimTextOutput(outputId = "value_Group1"), - - #p("compared against"), - - #selection for group 2 (denominator) of the contrasts - #initially disabled to allow - #disabled( - selectizeInput(inputId = "Group2", - label = "Contrast Group 2", - choices = list( - Tregs = results_names[ - grepl(pattern = "cellTypeT", - x = results_names, - ignore.case = T)], - Tconv = results_names[ - grepl(pattern = "cellTypeNT", - x = results_names, - ignore.case = T)] - ), - selected = NULL, - #selectize = T, - multiple = T, - options = list( - placeholder = paste0("Click here ", - "to manually", - " select ", - "...") - ) - ), - - #fluidRow( - #h5("Select available:"), - #h5("Quick add available:"), - # actionButton(inputId = "Group2SelectTconv", - # label = "Tconv"), - # actionButton(inputId = "Group2SelectTregs", - # label = "Tregs"), - selectizeInput(inputId = "Group2Selector", - label = "Add available:", - choice = list( - Treg = c("All Tregs", - "Blood Tregs", - "Lymphoid Tregs", - "Non-lymphoid Tregs", - "Gut-associated Tregs"), - Tconv = c("All Tconv", - "Blood Tconv", - "Lymphoid Tconv", - "Non-lymphoid Tconv", - "Gut-associated Tconv") - ), - selected = "All Tconv", - multiple = F, - options = list( - placeholder = "Click to select") - ), - actionButton(inputId = "Group2Add", - label = "Add"), - actionButton(inputId = "Group2Clear", - label = "Clear Selection"), - #), - - hr(), - - checkboxInput(inputId = "AddOps", - label = "Show Additional Options"), - includeScript(path = "js/validate-number.js"), - conditionalPanel( - condition = "input.AddOps", - selectizeInput(inputId = "TPCFilt", - label = "Minimum Transcripts per Cell", - choice = c("No filter", "0.01", - "0.1", "1"), - selected = "0.01", - multiple = F), - h5("Volcano Plot Marking Criteria"), - numericInput(inputId = "log2FCFilt", - label = HTML(paste0("log2 Fold ", - "Change (+/-)", - "
(Range: ", - "0 - 20)")), - value = 2, - min = 0, - max = 20, - step = 0.1), - numericInput(inputId = "AdjpFilt", - label = "Adjusted p.value", - value = 0.01, - min = 0, - max = 1, - step = 0.01), - ), - hr(), - #output of group 2 selection for debugging -- remove/ - # hide when deploying - #verbatimTextOutput(outputId = "value_Group2"), - - #action button for starting the DESeq2 results() - # function for generating contrasts - actionButton(inputId = "GenContrasts", - label = "Generate Contrasts" - ), - - ), - - # verbatimTextOutput(outputId = "res_error"), - # - # h6("Debug console-like thingy"), - # verbatimTextOutput(outputId = "debug_console") - - ), - - column(width = 4, - - conditionalPanel( - condition = "output.show_panels", - - #h5(HTML("
")), - br(), - fluidRow( - column(width = 10, - p(paste0("For an overview of the results, ", - "features and interactability, please ", - "click the yellow button to the right.")) - ), - column(width = 2, - # actionButton(inputId = "TutorialButton2", - # label = "Start tutorial"), - actionBttn(inputId = "TutorialButton2", - label = "Start tutorial", - style = "material-circle", - color = "warning", - icon = icon("question")), - ) - ), - - hr(), - - checkboxInput(inputId = "PlotSaveOps", - label = "Show Plot Export Options"), - - conditionalPanel( - condition = "input.PlotSaveOps", - - fluidRow( - column(width = 3,#) - radioGroupButtons(inputId = "SavePlotFormat", - label = HTML(paste0("Plot", - " Export ", - "Format")), - choices = c("SVG", "PNG", "JPEG"), - #SVG, PNG, JPEG and WEBP - #np native PDF support - selected = "SVG", - direction = "vertical", #"horizontal", - checkIcon = list( - # yes = icon("ok", - # lib = "glyphicon") - yes = icon(name = "file-image", - lib = "font-awesome") - ), - size = "normal" # xs/sm/normal/lg - ), - ), - column(width = 7,#) - includeScript(path = "js/validate-number.js"), - - numericInputIcon(inputId = "WidthPixels", - label = HTML(paste0("Plot ", - "Size in ", - " Pixels ", #"
", - "(Range: 100 ", - "- 10,000) ", - #"

", - "
", - "Width")), #in Pixels", - value = 880, - min = 100, - max = 10000, - step = 10, - width = "85%", - icon = list( - icon(name = "ruler-horizontal", - lib = "font-awesome"), - "px") - ), - - numericInputIcon(inputId = "HeightPixels", - label = "Height", #in Pixels", - value = 562, - min = 100, - max = 10000, - step = 10, - width = "85%", - icon = list( - icon(name = "ruler-vertical", - lib = "font-awesome"), - "px") - ) - ) - ) - ), - - hr(), - - h4("Differential Expression Volcano Plot"), - # h5(paste0("Click on data points to select/deselect ", - # "them in the table and show/hide their ", - # "normalised counts.")), - - plotlyOutput(outputId = "res_volplot") %>% - withSpinner(type = 3, size = 0.5, - color.background = "#FFFFFF"), - ), - - conditionalPanel( - condition = paste0( - "typeof input.res_table_rows_selected !== ", - "'undefined' && ", - "input.res_table_rows_selected.length > 0" - ), - hr(), - h4("Normalised Expression Plot"), - - plotlyOutput(outputId = "res_colplot") %>% - withSpinner(type = 3, size = 0.5, - color.background = "#FFFFFF") - ) - - ), - - column(width = 5, - - fluidRow( - conditionalPanel( - condition = "output.show_panels", - - #fluidRow( - #column(width = 11, - h4(HTML("
Results Table")), - h5("Download tab delimited file:"), - downloadButton( - outputId = "DownloadTableAll", - label = "Complete Results" - ), - downloadButton( - outputId = "DownloadTableFilt", - label = "Filtered Table" - ), - downloadButton( - outputId = "DownloadTableSelected", - label = "Selected Rows Only" - ) - #) - #), - ), - ), - #br(), - conditionalPanel( - condition = "output.show_panels", - fluidRow( - column(width = 12, - hr(), - #h5("Row Selection Options:"), - actionButton(inputId = "ResetTable", - label = "Reset Row Selection" - ) - ) - ), - br(), - ), - - # conditionalPanel( - # condition = paste0( - # "typeof output.res_table !== 'undefined'" - # ), - #shinyjs::hidden( - fluidRow( - #shinyjs::hidden( - DT::dataTableOutput(outputId = "res_table") #%>% - # withSpinner(type = 3, size = 0.5, - # color.background = "#FFFFFF") - #) - ) - ) - ) - ) - }, - - #Pathway analysis panel - { - tabPanel(title = "Pathway Analysis", value = "PathwayAnalysis", - - h3("Visualise enriched KEGG pathways"), - - p(paste0("The results of the pathway analsis using GAGE and ", - "Pathview can be visualised below.")), - - hr(), - - #vr(), - - column(width = 9, - - fluidRow( - - #h4("Visualise Enriched KEGG Pathways"), - - column(width = 6, - #br(), - br(), - - selectizeInput(inputId = "PathviewPlotSelect", - label = "Select KEGG Pathway", - # choices = as.list(results_names), - #choices = c("", "PCA", "t-SNE", "UMAP"), - choices = paste(Pathview_mapping[,1], - Pathview_mapping[,2], - sep = " - "), - selected = paste(Pathview_mapping[,1], - Pathview_mapping[,2], - sep = " - ")[1], - #selectize = T, - multiple = F, - options = list( - placeholder = "Click to select"), - width = "600px" - ), - - downloadButton( - outputId = "DownloadPathviewSelected", - label = "Selected Pathway" - ), - - downloadButton( - outputId = "DownloadPathviewAll", - label = "All Pathways" - ) - - ), - - column(width = 3, - - imageOutput(outputId = "PathviewPlotLegend", - width = "100%", - height = "100%") %>% - withSpinner(type = 3, size = 0.5, - color.background = "#FFFFFF"), - br() - ), - - column(width = 3, - - br(), - imageOutput(outputId = "PathviewPlotLegend2", - width = "100%", - height = "100%") %>% - withSpinner(type = 3, size = 0.5, - color.background = "#FFFFFF") - )#, - ), - - imageOutput(outputId = "PathviewPlotImage", - width = 1342*0.35, - height = 1451*0.35) %>% - withSpinner(type = 3, size = 0.5, - color.background = "#FFFFFF") - - ) - ) - - }, - - #Single Cell Expression panel - # { - # tabPanel(title = "Single Cell Expression", value = "SingleCellExprs", - # - # ) - # }, - - #TCR panel - { - tabPanel(title = "TCR", value = "TCR", - - h3("TCR repertoire analysis"), - - p(paste0("Description of the TCR repertoire analysis - ", - "ask Orian")), - - #br(), - hr(), - - fluidRow( - - column(width = 4, - - selectizeInput(inputId = "TCRChordSelect", - label = "Select Tissue", - # choices = as.list(results_names), - #choices = c("", "PCA", "t-SNE", "UMAP"), - choices = gsub(pattern = "_.*", - replacement = "", - x = TCR_plots_list, - ignore.case = T), - selected = "Blood", - #selectize = T, - multiple = F, - options = list( - placeholder = "Click to select") - ), - - downloadButton( - outputId = "DownloadTCRSelected", - label = "Selected Tissue" - ), - - downloadButton( - outputId = "DownloadTCRAll", - label = "All Tissues" - ) - - ), - - column(width = 8, - - imageOutput(outputId = "TCRChordImage", - width = 4344*0.15, - height = 4601*0.15) %>% - withSpinner(type = 3, size = 0.5, - color.background = "#FFFFFF") - - ) - - ) - ) - } - - - ), - ) - ## ui fluid page ends at bracket immediately above - -} - - -#shiny app server -{ - - # Define server logic ---- - server <- function(input, output, session) { - - #reading inputs for debugging - #output$value_Group1 <- renderPrint({ (input$Group1) }) - #output$value_Group2 <- renderPrint({ (input$Group2) }) - - #output$value_padj <- renderPrint({ (input$Padj_cutoff) }) - - #debug console output - # output$debug_console <- renderPrint(results_names[ - # grepl(pattern = "_NT", - # x = results_names, - # ignore.case = T)]) - # output$debug_console <- renderPrint(class(input$Group1)) - - #external variable handles so that they do not get destroyed once relevant - #gui updates and rendering are performed - global_data <- reactiveValues(result = NULL, - base_traces = 0, - #plotly traces are 0 indexed so count is ct-1 - trace_count = 0, - #trace_count = base_traces-1, - dt = NULL, - #volplot_selected = NULL#, - #volplot = NULL, - #colplot = NULL - left_log = 0, - right_log = 0, - first_run_done = F - ) - - #live updating UI elements - observe({ - - updateSelectInput(session = session, - inputId = "Group1", - label = "Contrast Group 1", - # choices = as.list(results_names[ - # !(results_names %in% input$Group2) - # ]), - choices = list( - Tregs = results_names[ - grepl(pattern = "cellTypeT", - x = results_names, - ignore.case = T) & !(results_names %in% - input$Group2)], - Tconv = results_names[ - grepl(pattern = "cellTypeNT", - x = results_names, - ignore.case = T) & !(results_names %in% - input$Group2)] - ), - selected = input$Group1) - - updateSelectInput(session = session, - inputId = "Group2", - label = "Contrast Group 2", - # choices = as.list(results_names[ - # !(results_names %in% input$Group1) - # ]), - choices = list( - Tregs = results_names[ - grepl(pattern = "cellTypeT", - x = results_names, - ignore.case = T) & !(results_names %in% - input$Group1)], - Tconv = results_names[ - grepl(pattern = "cellTypeNT", - x = results_names, - ignore.case = T) & !(results_names %in% - input$Group1)] - ), - selected = input$Group2) - - updateNumericInput(session = session, - inputId = "HeightPixels", - # label = tags$div(HTML(paste0(' Plot ', - # 'Height (pixels)') - # ) - # ), - value = ifelse(!is.numeric(input$HeightPixels), 562, - ifelse(input$HeightPixels > 10000, - 10000, ifelse( - input$HeightPixels < 100, 100, - round(input$HeightPixels)) - ) - ), - min = 100, - max = 10000, - step = 10 - ) - - updateNumericInput(session = session, - inputId = "WidthPixels", - # label = tags$div(HTML(paste0(' Plot ', - # 'Width (pixels)') - # ) - # ), - value = ifelse(!is.numeric(input$WidthPixels), 880, - ifelse(input$WidthPixels > 10000, - 10000, ifelse( - input$WidthPixels < 100, 100, - round(input$WidthPixels)) - ) - ), - min = 100, - max = 10000, - step = 10 - ) - - updateNumericInput(session = session, - inputId = "log2FCFilt", - value = ifelse(input$log2FCFilt > 20, 20, - ifelse(input$log2FCFilt < 0, - -(input$log2FCFilt), - ifelse( - !is.numeric(input$log2FCFilt) | - is.na(input$log2FCFilt) | - is.null(input$log2FCFilt) | - input$log2FCFilt == "" | - input$log2FCFilt == 0, - 2, input$log2FCFilt) - ) - ), - min = 0, max = 20, step = 0.1 - ) - updateNumericInput(session = session, - inputId = "AdjpFilt", - value = ifelse(input$AdjpFilt > 1, 1, - ifelse(input$AdjpFilt < 0, - -(input$AdjpFilt), - ifelse( - !is.numeric(input$AdjpFilt) | - is.na(input$AdjpFilt) | - is.null(input$AdjpFilt) | - input$AdjpFilt == "" | - input$AdjpFilt == 0, - 0.01, input$AdjpFilt) - ) - ), - min = 0, max = 1, step = 0.01 - ) - #print(input$AdjpFilt) - - }) - - #update available samples in contrast group 1 selection buttons - { - #adding entries to selection based on group - observeEvent(input$Group1Add, { - group1_selection <- unlist(str_split(string = input$Group1Selector, - pattern = " ")) - - #print(results_names) - #print(group1_selection) - if(group1_selection[2] == "Tregs"){ - available <- results_names[grepl(pattern = "cellTypeT", - x = results_names, - ignore.case = T)] - } - else{ - available <- results_names[grepl(pattern = "cellTypeNT", - x = results_names, - ignore.case = T)] - } - #print(available) - - if(group1_selection[1] == "Blood"){ - updateSelectInput(session = session, - inputId = "Group1", - label = "Contrast Group 1", - choices = list( - Tregs = results_names[ - grepl(pattern = "cellTypeT", - x = results_names, - ignore.case = T) & !(results_names %in% - input$Group2)], - Tconv = results_names[ - grepl(pattern = "cellTypeNT", - x = results_names, - ignore.case = T) & !(results_names %in% - input$Group2)] - ), - selected = c(input$Group1, available[ - grepl(pattern = "Blood", - x = available, - ignore.case = T) & !(available %in% - input$Group2)]) - ) - } - else if(group1_selection[1] == "Lymphoid"){ - updateSelectInput(session = session, - inputId = "Group1", - label = "Contrast Group 1", - choices = list( - Tregs = results_names[ - grepl(pattern = "cellTypeT", - x = results_names, - ignore.case = T) & !(results_names %in% - input$Group2)], - Tconv = results_names[ - grepl(pattern = "cellTypeNT", - x = results_names, - ignore.case = T) & !(results_names %in% - input$Group2)] - ), - selected = c(input$Group1, available[ - grepl(pattern = "(LymphNode|Spleen|PeyersPatches)", - x = available, - ignore.case = T) & !(available %in% - input$Group2)]) - ) - } - else if(group1_selection[1] == "Non-lymphoid"){ - updateSelectInput(session = session, - inputId = "Group1", - label = "Contrast Group 1", - choices = list( - Tregs = results_names[ - grepl(pattern = "cellTypeT", - x = results_names, - ignore.case = T) & !(results_names %in% - input$Group2)], - Tconv = results_names[ - grepl(pattern = "cellTypeNT", - x = results_names, - ignore.case = T) & !(results_names %in% - input$Group2)] - ), - selected = c(input$Group1, available[ - grepl(pattern = "(Kidney|Liver|Lung|Pancreas)", - x = available, - ignore.case = T) & !(available %in% - input$Group2)]) - ) - } - else if(group1_selection[1] == "Gut-associated"){ - updateSelectInput(session = session, - inputId = "Group1", - label = "Contrast Group 1", - choices = list( - Tregs = results_names[ - grepl(pattern = "cellTypeT", - x = results_names, - ignore.case = T) & !(results_names %in% - input$Group2)], - Tconv = results_names[ - grepl(pattern = "cellTypeNT", - x = results_names, - ignore.case = T) & !(results_names %in% - input$Group2)] - ), - selected = c(input$Group1, available[ - grepl(pattern = "(IntraEpithelialLayer|LaminaPropriaLymphocyte)", - x = available, - ignore.case = T) & !(available %in% - input$Group2)]) - ) - } - else{ - updateSelectInput(session = session, - inputId = "Group1", - label = "Contrast Group 1", - choices = list( - Tregs = results_names[ - grepl(pattern = "cellTypeT", - x = results_names, - ignore.case = T) & !(results_names %in% - input$Group2)], - Tconv = results_names[ - grepl(pattern = "cellTypeNT", - x = results_names, - ignore.case = T) & !(results_names %in% - input$Group2)] - ), - selected = c(input$Group1, available[ - !(available %in% input$Group2)]) - ) - } - - }) - - #clear selection - observeEvent(input$Group1Clear, { - - updateSelectInput(session = session, - inputId = "Group1", - label = "Contrast Group 1", - choices = list( - CD4s = results_names[ - grepl(pattern = "cellTypeNT", - x = results_names, - ignore.case = T) & !(results_names %in% - input$Group2)], - Tregs = results_names[ - grepl(pattern = "cellTypeT", - x = results_names, - ignore.case = T) & !(results_names %in% - input$Group2)] - ), - selected = NULL - ) - - }) - - #old button code - { - # #all CD4s - # observeEvent(input$Group1SelectTconv, { - # - # updateSelectInput(session = session, - # inputId = "Group1", - # label = "Contrast Group 1", - # choices = list( - # CD4s = results_names[ - # grepl(pattern = "cellTypeNT", - # x = results_names, - # ignore.case = T) & !(results_names %in% - # input$Group2)], - # Tregs = results_names[ - # grepl(pattern = "cellTypeT", - # x = results_names, - # ignore.case = T) & !(results_names %in% - # input$Group2)] - # ), - # selected = c(input$Group1, results_names[ - # grepl(pattern = "cellTypeNT", - # x = results_names, - # ignore.case = T) & !(results_names %in% - # input$Group2)]) - # ) - # - # }) - # - # #all Tregs - # observeEvent(input$Group1SelectTregs, { - # - # updateSelectInput(session = session, - # inputId = "Group1", - # label = "Contrast Group 1", - # choices = list( - # CD4s = results_names[ - # grepl(pattern = "cellTypeNT", - # x = results_names, - # ignore.case = T) & !(results_names %in% - # input$Group2)], - # Tregs = results_names[ - # grepl(pattern = "cellTypeT", - # x = results_names, - # ignore.case = T) & !(results_names %in% - # input$Group2)] - # ), - # selected = c(input$Group1, results_names[ - # grepl(pattern = "cellTypeT", - # x = results_names, - # ignore.case = T) & !(results_names %in% - # input$Group2)]) - # ) - # - # }) - # - # #all remaining - # # observeEvent(input$Group1SelectAll, { - # # - # # updateSelectInput(session = session, - # # inputId = "Group1", - # # label = "Contrast Group 1", - # # choices = list( - # # CD4s = results_names[ - # # grepl(pattern = "cellTypeNT", - # # x = results_names, - # # ignore.case = T) & !(results_names %in% - # # input$Group2)], - # # Tregs = results_names[ - # # grepl(pattern = "cellTypeT", - # # x = results_names, - # # ignore.case = T) & !(results_names %in% - # # input$Group2)] - # # ), - # # selected = c(input$Group1, results_names[ - # # grepl(pattern = "cellTypeNT", - # # x = results_names, - # # ignore.case = T) & !(results_names %in% - # # input$Group2)], - # # results_names[ - # # grepl(pattern = "cellTypeT", - # # x = results_names, - # # ignore.case = T) & !(results_names %in% - # # input$Group2)]) - # # ) - # # - # # }) - # - } - - } - - #update available samples in contrast group 2 selection buttons - { - #adding entries to selection based on group - observeEvent(input$Group2Add, { - group2_selection <- unlist(str_split(string = input$Group2Selector, - pattern = " ")) - #print(group1_selection) - if(group2_selection[2] == "Tregs"){ - available <- results_names[grepl(pattern = "cellTypeT", - x = results_names, - ignore.case = T)] - } - else{ - available <- results_names[grepl(pattern = "cellTypeNT", - x = results_names, - ignore.case = T)] - } - #print(available) - - if(group2_selection[1] == "Blood"){ - updateSelectInput(session = session, - inputId = "Group2", - label = "Contrast Group 2", - choices = list( - Tregs = results_names[ - grepl(pattern = "cellTypeT", - x = results_names, - ignore.case = T) & !(results_names %in% - input$Group1)], - Tconv = results_names[ - grepl(pattern = "cellTypeNT", - x = results_names, - ignore.case = T) & !(results_names %in% - input$Group1)] - ), - selected = c(input$Group2, available[ - grepl(pattern = "Blood", - x = available, - ignore.case = T) & !(available %in% - input$Group1)]) - ) - } - else if(group2_selection[1] == "Lymphoid"){ - updateSelectInput(session = session, - inputId = "Group2", - label = "Contrast Group 2", - choices = list( - Tregs = results_names[ - grepl(pattern = "cellTypeT", - x = results_names, - ignore.case = T) & !(results_names %in% - input$Group1)], - Tconv = results_names[ - grepl(pattern = "cellTypeNT", - x = results_names, - ignore.case = T) & !(results_names %in% - input$Group1)] - ), - selected = c(input$Group2, available[ - grepl(pattern = "(LymphNode|Spleen|PeyersPatches)", - x = available, - ignore.case = T) & !(available %in% - input$Group1)]) - ) - } - else if(group2_selection[1] == "Non-lymphoid"){ - updateSelectInput(session = session, - inputId = "Group2", - label = "Contrast Group 2", - choices = list( - Tregs = results_names[ - grepl(pattern = "cellTypeT", - x = results_names, - ignore.case = T) & !(results_names %in% - input$Group1)], - Tconv = results_names[ - grepl(pattern = "cellTypeNT", - x = results_names, - ignore.case = T) & !(results_names %in% - input$Group1)] - ), - selected = c(input$Group2, available[ - grepl(pattern = "(Kidney|Liver|Lung|Pancreas)", - x = available, - ignore.case = T) & !(available %in% - input$Group1)]) - ) - } - else if(group2_selection[1] == "Gut-associated"){ - updateSelectInput(session = session, - inputId = "Group2", - label = "Contrast Group 2", - choices = list( - Tregs = results_names[ - grepl(pattern = "cellTypeT", - x = results_names, - ignore.case = T) & !(results_names %in% - input$Group1)], - Tconv = results_names[ - grepl(pattern = "cellTypeNT", - x = results_names, - ignore.case = T) & !(results_names %in% - input$Group1)] - ), - selected = c(input$Group2, available[ - grepl(pattern = "(IntraEpithelialLayer|LaminaPropriaLymphocyte)", - x = available, - ignore.case = T) & !(available %in% - input$Group1)]) - ) - } - else{ - updateSelectInput(session = session, - inputId = "Group2", - label = "Contrast Group 2", - choices = list( - Tregs = results_names[ - grepl(pattern = "cellTypeT", - x = results_names, - ignore.case = T) & !(results_names %in% - input$Group1)], - Tconv = results_names[ - grepl(pattern = "cellTypeNT", - x = results_names, - ignore.case = T) & !(results_names %in% - input$Group1)] - ), - selected = c(input$Group2, available[ - !(available %in% input$Group1)]) - ) - } - - }) - - #clear selection - observeEvent(input$Group2Clear, { - - updateSelectInput(session = session, - inputId = "Group2", - label = "Contrast Group 2", - choices = list( - CD4s = results_names[ - grepl(pattern = "cellTypeNT", - x = results_names, - ignore.case = T) & !(results_names %in% - input$Group1)], - Tregs = results_names[ - grepl(pattern = "cellTypeT", - x = results_names, - ignore.case = T) & !(results_names %in% - input$Group1)] - ), - selected = NULL - ) - - }) - - #old button code - { - # #all CD4s - # observeEvent(input$Group2SelectTconv, { - # - # updateSelectInput(session = session, - # inputId = "Group2", - # label = "Contrast Group 2", - # choices = list( - # CD4s = results_names[ - # grepl(pattern = "cellTypeNT", - # x = results_names, - # ignore.case = T) & !(results_names %in% - # input$Group1)], - # Tregs = results_names[ - # grepl(pattern = "cellTypeT", - # x = results_names, - # ignore.case = T) & !(results_names %in% - # input$Group1)] - # ), - # selected = c(input$Group2, results_names[ - # grepl(pattern = "cellTypeNT", - # x = results_names, - # ignore.case = T) & !(results_names %in% - # input$Group1)]) - # ) - # - # }) - # - # #all Tregs - # observeEvent(input$Group2SelectTregs, { - # - # updateSelectInput(session = session, - # inputId = "Group2", - # label = "Contrast Group 2", - # choices = list( - # CD4s = results_names[ - # grepl(pattern = "cellTypeNT", - # x = results_names, - # ignore.case = T) & !(results_names %in% - # input$Group1)], - # Tregs = results_names[ - # grepl(pattern = "cellTypeT", - # x = results_names, - # ignore.case = T) & !(results_names %in% - # input$Group1)] - # ), - # selected = c(input$Group2, results_names[ - # grepl(pattern = "cellTypeT", - # x = results_names, - # ignore.case = T) & !(results_names %in% - # input$Group1)]) - # ) - # - # }) - # - # #all remaining - # # observeEvent(input$Group2SelectAll, { - # # - # # updateSelectInput(session = session, - # # inputId = "Group2", - # # label = "Contrast Group 2", - # # choices = list( - # # CD4s = results_names[ - # # grepl(pattern = "cellTypeNT", - # # x = results_names, - # # ignore.case = T) & !(results_names %in% - # # input$Group1)], - # # Tregs = results_names[ - # # grepl(pattern = "cellTypeT", - # # x = results_names, - # # ignore.case = T) & !(results_names %in% - # # input$Group1)] - # # ), - # # selected = c(input$Group2, results_names[ - # # grepl(pattern = "cellTypeNT", - # # x = results_names, - # # ignore.case = T) & !(results_names %in% - # # input$Group1)], - # # results_names[ - # # grepl(pattern = "cellTypeT", - # # x = results_names, - # # ignore.case = T) & !(results_names %in% - # # input$Group1)]) - # # ) - # # - # # }) - # - } - - } - - #enabling / disabling UI elements - observe({ - shinyjs::toggleState(id = "GenContrasts", - condition = !(length(input$Group1) < 1 | - length(input$Group2) < 1) && - (is.numeric(input$log2FCFilt) & - is.numeric(input$AdjpFilt)) - ) - }) - #output$show_panels <- reactive({FALSE}) - #outputOptions(x = output, name = "show_panels", suspendWhenHidden = FALSE) - shinyjs::hide(id = "res_table") - #shinyjs::disable(id = "res_table") - - - #generating contrasts via the DESeq2 results() function - observeEvent(input$GenContrasts, { - - #splash alert for the user informing them that the analysis may take some - # time - if(!global_data$first_run_done){ - # sendSweetAlert(session = session, - # title = "Computing contrasts", - # text = paste0("The computation may take some time during ", - # "which the tool may appear to not be doing ", - # "anything. Once the results are computed ", - # "they will populate the tab."), - # type = "info") - shinyalert(title = "Computing contrasts", - text = paste0("The computation may take some time during ", - "which the tool may appear to not be doing ", - "anything. Once the results are computed, ", - "they will populate the tab."), - type = "info", - animation = F, #"slide-from-bottom", - inputId = "AlertFirstRunDone") - global_data$first_run_done <- T - } - - - #ensuring that the minimum log2FC and minimum adj.p.value fields weren't - # left blank for the fraction of the second it takes to disable the - # generate contrast button - req(input$log2FCFilt, input$AdjpFilt, input$TPCFilt) - - #disabling the GenContrasts button so that users may not spam it - #is renabled at the end of this observeEvent block - shinyjs::disable(id = "GenContrasts") - #enabling the results table rendered object - #shinyjs::show(id = "res_table") - - #resetting the previous output if it exists - output$res_error <- NULL - output$res_table <- NULL - output$res_volplot <- NULL - output$res_colplot <- NULL - - #print(input$TPCFilt) - #checking which filter list to send to the contrast - if(input$TPCFilt == "No filter"){ - tpc_filtered_list <- NULL - } - else{ - tpc_filtered_list <- filtered_genes_lists[[paste0("Filt", - input$TPCFilt)]] - } - - #performing the contrast - result <- GenerateContrasts(DESeq_object = DESeq_object, - group1 = input$Group1, - group2 = input$Group2, - meta_data = MGI_info, - mean_norm_counts = mean_norm_counts, - tpc_filtered_list = tpc_filtered_list - ) - #print(class(result[[2]])) - #global_data$result <- result - - #filtering results for only genes for which actual logFC exists - # actually bad idea because we would lose genes which are not being - # in one contrast group (e.g., those which are only expressed in Treg and - # not Tconv) - # actual_logFC <- - # - # (is.finite(log2(rowMeans(norm_counts[,2:4])) - log2(rowMeans(norm_counts[,6:8]))))[order()] - - #interpretting the type of output and updating relevant UI elements - if(length(result) == 2){ - - #outputting error - output$res_error <- renderText({ result[[1]] }) - - #saving the results in a shared variable - global_data$result <- result[[2]] - - result <- result[[2]] - - } - else if(class(result) == "data.frame"){ - #saving the results in a shared variable - global_data$result <- result - - } - else if(class(result) == "character"){ - output$res_error <- renderText({ result }) - } - - #print(summary(isolate(global_data$result))) - #calculating the fold change values for the left and right flanking - # columns of the NA/NaN/Inf/-Inf genes in the volcano plot - # --can be sent to the plotting function too, but I already have that up - # -- and running; this is for the table/data point selection and traces - global_data$left_log <- -(ceiling(max(abs( - isolate(global_data$result$DESeq2.Beta.Coefficients[ - is.finite(isolate(global_data$result$log2.Fold.Change))])))) + 5) #* -1 - #))) - #print(isolate(global_data$left_log)) - global_data$right_log <- (ceiling(max(abs( - isolate(global_data$result$DESeq2.Beta.Coefficients[ - is.finite(isolate(global_data$result$log2.Fold.Change))])))) + 5) - #))) - # print(isolate(global_data$right_log)) - - - #processing data frame to be displayed as DataTable - if(class(result) == "data.frame"){ - - #replacing zero p.values/adj.p.values with smallest IEEE supported value - global_data$result$p.value[global_data$result$p.value == 0] <- - .Machine$double.xmin - global_data$result$Adjusted.p.value[ - global_data$result$Adjusted.p.value == 0] <- .Machine$double.xmin - - result <- global_data$result - - #removing extra columns - result$Standard.Error <- NULL - result$Statistic <- NULL - result$p.value <- NULL - - #formatting the numeric columns out here rather than through DT::format* - # so that the filter ranges do not end up with 15 decimal places - result$Mean.Counts <- round(x = result$Mean.Counts, digits = 3) - result$log2.Fold.Change <- round(x = result$log2.Fold.Change, - digits = 3) - result$DESeq2.Beta.Coefficients <- - round(x = result$DESeq2.Beta.Coefficients, digits = 3) - result$Adjusted.p.value <- signif(x = result$Adjusted.p.value, - digits = 3) - - #adding selection column for sorting selected rows -- not being used atm - result$Selected <- "No" - - #outputting results - output$res_table <- DT::renderDataTable({ - DT::datatable(elementId = "res_table", - data = result, - #caption = "Differential expression results", - colnames = gsub(pattern = "\\.", - replacement = " ", - x = colnames(result), - ignore.case = T), - #extensions = "Buttons", - options = list(searching = T, - searchHighlight = TRUE, - dom = "lrtip", - #buttons = c('csv', 'excel', 'pdf'), - #order = list(list(5, 'asc')) - order = list(list(6, 'asc')) - ), - filter = list(position = "top", - plain = F - ), - editable = F, - rownames = F) #%>% - # DT::formatRound(columns = c("Mean.Counts"), - # digits = 2, - # mark = "") %>% - # DT::formatRound(columns = c("log2.Fold.Change"), - # digits = 3, - # mark = "") %>% - # DT::formatSignif(columns = c("Adjusted.p.value"), - # digits = 3, - # mark = "") - }) - - #used for updating DataTable values for the 'Selected' column - global_data$dt <- result - - } - - - #checking if a contrast was generated to create the volcano plot - if(class(result) == "data.frame"){ - - #interpreting the label names for the plot title - names_group1 <- names(results_names[results_names %in% - isolate(input$Group1)]) - names_group2 <- names(results_names[results_names %in% - isolate(input$Group2)]) - - #turning off warning to hide the persistent "Warning: 'scattergl' - # objects don't have these attributes: 'hoveron'" warnings. Comment the - # below code when debugging the plotting code - #restoreWarn <- getOption("warn") - #options(warn = -1) - - #generating the ggplotly plot - # output$res_volplot <- renderPlotly({ - # GenerateVolcanoPlot(plot_data = isolate(global_data$result), - # names_group1 = names_group1, - # names_group2 = names_group2, - # format = tolower(isolate(input$SavePlotFormat)), - # width = isolate(input$WidthPixels), - # height = isolate(input$HeightPixels) - # ) - p <- GenerateVolcanoPlot(plot_data = isolate(global_data$result), - names_group1 = names_group1, - names_group2 = names_group2, - format = tolower(isolate(input$SavePlotFormat)), - width = isolate(input$WidthPixels), - height = isolate(input$HeightPixels), - log2FCFilt = input$log2FCFilt, - AdjpFilt = input$AdjpFilt - ) - output$res_volplot <- renderPlotly({p}) - p <- plotly_build(p) - global_data$base_traces <- length(p$x$data) - global_data$trace_count <- global_data$base_traces-1 - #plotly traces are zero indexed so trace_count is base_traces-1 - - #restoring warnings - #options(warn = restoreWarn) - - } - - #renabling the disabled GenContrasts button - shinyjs::enable(id = "GenContrasts") - #shinyjs::enable(id = "res_table") - - #showing hidden ui controls and elements - shinyjs::show(id = "res_table") - output$show_panels <- reactive({TRUE}) #for multiple conditionalPanels - outputOptions(x = output, name = "show_panels", suspendWhenHidden = FALSE) - - # #update the SavePlotFormat once more to allow saving the default SVG - # updateRadioGroupButtons(session = session, - # inputId = "SavePlotFormat", - # selected = isolate(input$SavePlotFormat)) - - #resuming the DT row selection observer - #res_table_row_observer$resume() - - }) - - - #generating bar plot of normalised counts for the genes of the rows selected - # in the rendered DataTable - observeEvent(input$res_table_rows_selected, ignoreNULL = F, { - - #resetting any existing plot - output$res_colplot <- NULL - - #remove any existing plotly traces on the volcano plot - # plotly traces are zero indexed so min count is global_data$base_traces-1 - #if(global_data$trace_count > 2){ - if(global_data$trace_count > global_data$base_traces-1){ - plotlyProxy(outputId = "res_volplot", session = session) %>% - plotlyProxyInvoke("deleteTraces", - #as.list(c(2:global_data$trace_count)) - as.list(c( - global_data$base_traces:global_data$trace_count)) - ) - - global_data$trace_count <- global_data$base_traces-1 - #print(global_data$trace_count) - } - - #if there is at least one row selected - if(!is.null(input$res_table_rows_selected)){ - - #hard limiting max selectable rows to first 50 selected - if(length(input$res_table_rows_selected) > 50){ - selected_rows <- input$res_table_rows_selected[1:50] - DT::dataTableProxy("res_table") %>% selectRows(selected_rows) - } - else{ - selected_rows <- input$res_table_rows_selected - } - - selection <- isolate(global_data$result[selected_rows, 1:2]) - - #generating the counts plot - output$res_colplot <- renderPlotly({ - GenerateColPlot(mean_norm_counts = mean_norm_counts, - selection = selection, - format = tolower(isolate(input$SavePlotFormat)), - width = isolate(input$WidthPixels), - height = isolate(input$HeightPixels) - ) - }) - - #adding traces to the plotly volcano plot - #need cols 5 and 9 from selection - selection <- isolate(global_data$result[selected_rows,]) - #print(summary(selection)) - - #colnames: 1-Gene.ID 2-Gene.Symbol 3-Description 4-Mean.Counts - # 5-log2.Fold.Change 6-DESeq2.Beta.Coefficients 7-p.value - # 8-Adjusted.p.value 9-Standard.Error 10-Statistic - - #output$debug_console <- renderPrint(selection) - - #plotly does not show traces for single items -- - #duplicating item if single - if(nrow(selection) == 1){ - #checking if log2.Fold.Change exists, then use DESeq2.Beta.Coefficients - # else use max/min for the right/left columns - if(is.finite(selection[1,5])){ - selected_x <- c(selection[1,6],selection[,6]) - } - else{ - if(selection[1,6] < 0) selected_x <- - c(isolate(global_data$left_log), isolate(global_data$left_log)) - else if(selection[1,6] > 0) selected_x <- - c(isolate(global_data$right_log), isolate(global_data$right_log)) - } - #selected_y <- c(selection[1,9],selection[,9]) - selected_y <- c(selection[1,8],selection[,8]) - - #if MGI or other symbols are missing - #if(selection[,2] == ""){ - if(is.na(selection[,2]) | selection[,2] == ""){ - # selected_text <- c(str_extract(selection[,1], '\\w*')[1], - # str_extract(selection[,1], '\\w*')) - selected_text <- c(gsub(pattern = ";.*", - replacement = "", - x = selection[,1], - ignore.case = T)[1], - gsub(pattern = ";.*", - replacement = "", - x = selection[,1], - ignore.case = T) - ) - #print(selected_text) - } - else{ - # selected_text <- c(str_extract(selection[,2], '\\w*')[1], - # str_extract(selection[,2], '\\w*')) - selected_text <- c(gsub(pattern = ";.*", - replacement = "", - x = selection[,2], - ignore.case = T)[1], - gsub(pattern = ";.*", - replacement = "", - x = selection[,2], - ignore.case = T) - ) - #print(selected_text) - } - } - else{ - #selected_x <- selection[,5] - selected_x <- selection[,6] - #checking if log2.Fold.Change exists, then use DESeq2.Beta.Coefficients - # else use max/min for the right/left columns - selected_x[!is.finite(selection[,5]) & selection[,6] < 0] <- - isolate(global_data$left_log) - selected_x[!is.finite(selection[,5]) & selection[,6] > 0] <- - isolate(global_data$right_log) - - #selected_y <- selection[,9] - selected_y <- selection[,8] - - #if MGI or other symbols are missing - #if(selection[,2] == ""){ - if(any(is.na(selection[,2])) | any(selection[,2] == "")){ - #print(selection[,2]) - #selected_text <- str_extract(selection[,1], '\\w*') - selected_text <- gsub(pattern = ";.*", - replacement = "", - x = selection[,1], - ignore.case = T) - selected_text[-(which(is.na(selection[,2]) | - selection[,2] == ""))] <- - gsub(pattern = ";.*", - replacement = "", - x = selection[-(which(is.na(selection[,2]) | - selection[,2] == "")),2], - ignore.case = T) - # selected_text[-(which(is.na(selection[,2]) | - # selection[,2] == ""))] <- - # str_extract(selection[-(which(is.na(selection[,2]) | - # selection[,2] == "")),2], '\\w*') - #print(selected_text) - } - else{ - #selected_text <- str_extract(selection[,2], '\\w*') - selected_text <- gsub(pattern = ";.*", - replacement = "", - x = selection[,2], - ignore.case = T) - } - } - - plotlyProxy(outputId = "res_volplot", session = session) %>% - plotlyProxyInvoke("addTrace", - list(#x = c(selection[1,5],selection[,5]), - #y = -log10(c(selection[1,9], selection[,9])), - x = selected_x, - y = -log10(selected_y), - type = "scattergl", - name = "Selected genes", - mode = "markers+text", - marker = list(#symbol = "x", - symbol = "circle-open", - size = 10, - #colorscale = "Viridis" - color = "blue" - ), - # text = c(str_extract(selection[,2], '\\w*')[1], - # str_extract(selection[,2], '\\w*')), - text = selected_text, - textposition = "top right" , - #textfont_size = 4, - textfont = list(family = "sans serif", - size = 10, - color = "blue" - ) - ) - ) - - # p <- plotlyProxy(outputId = "res_volplot", session = session)$x$data - # saveRDS(object = p, file = "p_proxy_build.rds") - - global_data$trace_count <- global_data$trace_count + 1 - #print(global_data$trace_count) - #output$debug_console <- renderPrint(global_data$trace_count) - - #output$debug_console <- renderPrint(selection) - - #update the selection column of the selected rows - #was designed so that selected rows can then be sorted on to move them to - #the top. however, the code currently invokes a perpetual looping update - #of the table and doesn't allow the selection of more than one row either - #probably due to reactivity of the row selection event -- maybe try - #binding to an action button instead of automatic updates via row - #selection? ---- code is now fixed - changes <- data.frame(row = selected_rows, - #col = 6, #column index of "selected" (index starts at 0) - col = 7, - value = "Yes") - - DT::dataTableProxy("res_table") %>% - DT::editData(data = isolate(global_data$dt), - info = changes, - rownames = F, - resetPaging = F, - clearSelection = "none") - - } - #else if the last row has been deselected, then reset selection column - else if(!is.null(input$res_table_row_last_clicked)){ - - changes <- data.frame(row = isolate(input$res_table_row_last_clicked), - #col = 6, #column index of "selected" (index starts at 0) - col = 7, - value = "No") - - DT::dataTableProxy("res_table") %>% - DT::editData(data = isolate(global_data$dt), - info = changes, - rownames = F, - resetPaging = F, - clearSelection = "all") - } - - }) - - - #resetting rows selected in the results table - observeEvent(input$ResetTable, { - proxy <- dataTableProxy("res_table") - - proxy %>% selectRows(NULL) - - output$res_colplot <- NULL - - #removing traces from the plotly volcano plot - # plotly traces are zero indexed so min count is global_data$base_traces-1 - if(global_data$trace_count > global_data$base_traces-1){ - plotlyProxy(outputId = "res_volplot", session = session) %>% - plotlyProxyInvoke("deleteTraces", - #as.list(c(2:global_data$trace_count)) - as.list(c( - global_data$base_traces:global_data$trace_count)) - ) - - global_data$trace_count <- global_data$base_traces-1 - #print(global_data$trace_count) - } - # if(global_data$trace_count > 2){ - # plotlyProxy(outputId = "res_volplot", session = session) %>% - # plotlyProxyInvoke("deleteTraces", - # #as.list(c(2:global_data$trace_count)) - # as.list(c(3:global_data$trace_count)) - # # as.list(c( - # # (global_data$trace_count-1):global_data$trace_count)) - # ) - # - # global_data$trace_count <- 2 - # } - - }) - - - #download the complete results - output$DownloadTableAll <- downloadHandler(filename = "Results.tab", - content = function(file){ - write.table( - x = global_data$result, - file = file, - quote = F, - sep = "\t", - col.names = T, - row.names = F) - }) - - #download the filtered results - output$DownloadTableFilt <- downloadHandler( - filename = "Results_filtered.tab", - content = function(file){ - write.table(x = - global_data$result[input$res_table_rows_all, , drop = F], - file = file, - quote = F, - sep = "\t", - col.names = T, - row.names = F) - }) - - #download the selected rows only - output$DownloadTableSelected <- downloadHandler( - filename = "Results_selected.tab", - content = function(file){ - write.table(x = global_data$result[ - input$res_table_rows_selected, , drop = F], - file = file, - quote = F, - sep = "\t", - col.names = T, - row.names = F) - }) - - #(de)select rows in the results table by (de)selecting it in volcano plot - observe({ - - #the req() circumvents the whole exists() and !is.null requirements in the - #outer if-else block below, reducing the check to just the 50 hard limit - req(event_data("plotly_click", - source = "volplot", - priority = "event")) - - volplot_selected <- event_data("plotly_click", - source = "volplot", - priority = "event") - proxy <- dataTableProxy("res_table") - - already_selected <- isolate(input$res_table_rows_selected) - # proxy %>% selectRows(c(volplot_selected$key, already_selected)) - - #the following code is designed to allow deselection, but it is bugged for - #some reason and requires the user to click the last selected node twice - #to deselect it will need to work on this in later updates if required as - #a feature ----- the bug is resolved by 'priority = "event"' above - #Also, hard limiting to max selections to 50 - # if(exists("volplot_selected") & !is.null(volplot_selected) & - # length(already_selected) < 50){ - if(length(already_selected) < 50){ - - if(length(intersect(volplot_selected$key, already_selected)) > 0){ - #print("entering 'if' block") - if(length(already_selected) > 1){ - #print("entering 'if>if' block") - proxy %>% selectRows(as.numeric( - setdiff(already_selected, volplot_selected$key))) - } - else{ - #print("entering 'if>else' block") - proxy %>% selectRows(NULL) - } - } - else if(length(union(volplot_selected$key, already_selected)) > 0){ - #print("entering 'else if' block") - proxy %>% selectRows(as.numeric( - union(volplot_selected$key, already_selected))) - } - # else if(length(volplot_selected$key) > 0 & - # length(already_selected) < 1){ - # proxy %>% selectRows(as.numeric(volplot_selected$key)) - # } - # else{ - # print("entering 'else' block") - # proxy %>% selectRows(NULL) - # } - - } - - #volplot_selected <- NULL - #print(isolate(global_data$trace_count)) - - }) - - - #creating reactive expression to group multiple inputs for the observeEvent - plotFormatAndSize <- reactive({ - list(input$SavePlotFormat, input$WidthPixels, input$HeightPixels) - }) - #change the snapshot save format and size of plotly plots - observeEvent(plotFormatAndSize(), { - - #formatting the input - format <- tolower(input$SavePlotFormat) - width <- input$WidthPixels - height <- input$HeightPixels - - #updating the format for the volcano plot - plotlyProxy(outputId = "res_volplot", session = session) %>% - plotlyProxyInvoke("reconfig", - displaylogo = F, - displayModeBar = T, - toImageButtonOptions = - list(format = format, - width = width, - height = height - ), - modeBarButtonsToRemove = list( - "sendDataToCloud", "zoom2d", "zoomIn2d", - "zoomOut2d", "pan2d", "select2d", "lasso2d", - "autoScale2d", "hoverClosestCartesian", - "hoverCompareCartesian" - ) - ) - - #updating the format for the counts plot - plotlyProxy(outputId = "res_colplot", session = session) %>% - plotlyProxyInvoke("reconfig", - displaylogo = F, - displayModeBar = T, - toImageButtonOptions = - list(format = format, - width = width, - height = height - ), - modeBarButtonsToRemove = list( - "sendDataToCloud", "zoom2d", "zoomIn2d", - "zoomOut2d", "pan2d", "select2d", "lasso2d", - "autoScale2d", "toggleSpikelines"#, - #"hoverClosestCartesian", - #"hoverCompareCartesian" - ) - ) - - }) - - - #display PCA, t-SNE and UMAP plots - observeEvent(input$TriplePlot, { - - output$PCAplot1 <- NULL - output$PCAplot2 <- NULL - output$PCAplot3 <- NULL - - # output$debug_console <- renderPrint(input$TriplePlot == "PCA") - - if(input$TriplePlot == "PCA"){ - #output$debug_console <- renderText(class(input$TriplePlot)) - #output$PCAplot1 <- renderPlotly({ - output$PCAplot1 <- renderPlot({ - #ggplotly(PCA_plots[[1]]) %>% toWebGL() - PCA_plots[[1]] - }) - output$PCAplot2 <- renderPlot({ - PCA_plots[[2]] - }) - output$PCAplot3 <- renderPlot({ - PCA_plots[[3]] - }) - } - else if(input$TriplePlot == "t-SNE"){ - output$PCAplot1 <- renderPlot({ - tSNE_plots[[1]] - }) - output$PCAplot2 <- renderPlot({ - tSNE_plots[[2]] - }) - output$PCAplot3 <- renderPlot({ - tSNE_plots[[3]] - }) - } - else if(input$TriplePlot == "UMAP"){ - output$PCAplot1 <- renderPlot({ - UMAP_plots[[1]] - }) - output$PCAplot2 <- renderPlot({ - UMAP_plots[[2]] - }) - output$PCAplot3 <- renderPlot({ - UMAP_plots[[3]] - }) - } - else if(input$TriplePlot == "PCA - tissue means"){ - output$PCAplot1 <- renderPlot({ - PCA_means_plots[[1]] - }) - output$PCAplot2 <- renderPlot({ - PCA_means_plots[[2]] - }) - output$PCAplot3 <- renderPlot({ - PCA_means_plots[[3]] - }) - } - - }) - - - #display TCR chord diagrams - observeEvent(input$TCRChordSelect, { - output$TCRChordImage <- renderImage(expr = { - - img <- readPNG( - source = paste0("www/TCR_plots/", - TCR_plots_list[grep(pattern = input$TCRChordSelect, - x = TCR_plots_list, - ignore.case = T)]), - native = T, info = T) - - list(src = paste0("www/TCR_plots/", - TCR_plots_list[grep(pattern = input$TCRChordSelect, - x = TCR_plots_list, - ignore.case = T)]), - contentType = 'image/png', - width = dim(img)[2]*0.15, - height = dim(img)[1]*0.15#, - #alt = "This is alternate text" - ) - }, deleteFile = F) - }) - - #display Pathview KEGG Pathways - observeEvent(input$PathviewPlotSelect, { - output$PathviewPlotImage <- renderImage(expr = { - - pathway_id <- gsub(pattern = " - .*", - replacement = "", - x = input$PathviewPlotSelect, - ignore.case = T) - - img <- readPNG( - source = paste0("www/Pathview_plots/", - Pathview_plots_list[grep(pattern = pathway_id, - x = Pathview_plots_list, - ignore.case = T)]), - native = T, info = T) - - list(src = paste0("www/Pathview_plots/", - Pathview_plots_list[grep(pattern = pathway_id, - x = Pathview_plots_list, - ignore.case = T)]), - contentType = 'image/png', - width = dim(img)[2],#*0.35, - height = dim(img)[1]#*0.35#, - #alt = "This is alternate text" - ) - }, deleteFile = F) - }) - - #display Pathview Legend and Legend2 - output$PathviewPlotLegend <- renderImage(expr = { - - img <- readPNG(source = "www/Legend.png", - native = T, info = T) - - list(src = "www/Legend.png", - contentType = 'image/png', - width = dim(img)[2], - height = dim(img)[1]#, - #alt = "This is alternate text" - ) - }, deleteFile = F) - output$PathviewPlotLegend2 <- renderImage(expr = { - - img <- readPNG(source = "www/Legend2.png", - native = T, info = T) - - list(src = "www/Legend2.png", - contentType = 'image/png', - width = dim(img)[2]*0.6, - height = dim(img)[1]*0.6#, - #alt = "This is alternate text" - ) - }, deleteFile = F) - - #download Pathview pathways - output$DownloadPathviewSelected <- downloadHandler( - filename = function() { - paste0(input$PathviewPlotSelect, ".zip") - }, - #filename = paste0(input$PathviewPlotSelect, ".zip"), - content = function(file){ - - pathway_id <- gsub(pattern = " - .*", - replacement = "", - x = input$PathviewPlotSelect, - ignore.case = T) - - zip(zipfile = file, - files = c(paste0("www/Pathview_plots/", - Pathview_plots_list[grep(pattern = pathway_id, - x = Pathview_plots_list, - ignore.case = T)]), - "www/Legend.png", "www/Legend2.png"), - flags = "-j") - }, - contentType = "application/zip") - # output$DownloadPathviewSelected <- downloadHandler( - # filename = paste0(input$PathviewPlotSelect, ".png"), - # content = function(file){ - # - # pathway_id <- gsub(pattern = " - .*", - # replacement = "", - # x = input$PathviewPlotSelect, - # ignore.case = T) - # - # file.copy(paste0("www/Pathview_plots/", - # Pathview_plots_list[grep(pattern = pathway_id, - # x = Pathview_plots_list, - # ignore.case = T)]), - # file, - # copy.mode = F, - # copy.date = F - # ) - # }, - # contentType = "image/png") - output$DownloadPathviewAll <- downloadHandler( - filename = function() { - paste0("all_pathways.zip") - }, - #filename = "all_pathways.zip", - content = function(file){ - - zip(zipfile = file, - files = c(paste0("www/Pathview_plots/", Pathview_plots_list), - "www/Legend.png", "www/Legend2.png"), - flags = "-j") - - }, - contentType = "application/zip") - - #download TCR plots - TCR_plots_list - output$DownloadTCRSelected <- downloadHandler( - filename = function() { - paste0(input$TCRChordSelect, "_chord_diagram.png") - }, - #filename = paste0(input$TCRChordSelect, "_chord_diagram.png"), - content = function(file){ - - file.copy(paste0("www/TCR_plots/", - TCR_plots_list[grep(pattern = input$TCRChordSelect, - x = TCR_plots_list, - ignore.case = T)]), - file, - copy.mode = F, - copy.date = F - ) - }, - contentType = "image/png") - output$DownloadTCRAll <- downloadHandler( - filename = function() { - paste0("all_chord_diagrams.zip") - }, - #filename = "all_chord_diagrams.zip", - content = function(file){ - - zip(zipfile = file, - files = paste0("www/TCR_plots/", TCR_plots_list), - flags = "-j") - - }, - contentType = "application/zip") - - #downloading Supplementary Materials - output$MultiQCBulk <- downloadHandler( - filename = function() { - paste0("QC and pre-processing report.zip") - }, - content = function(file){ - file.copy("www/MultiQC_report/QC and pre-processing report.zip", - file, - copy.mode = F, - copy.date = F - ) - }, - contentType = "application/zip") - output$GlobalBulk <- downloadHandler( - filename = function() { - paste0("PCA, t-SNE and UMAP plots.zip") - }, - content = function(file){ - zip(zipfile = file, - files = paste0("www/Global_bulk/", - list.files(path = "www/Global_bulk/")), - flags = "-j" - ) - }, - contentType = "application/zip") - #"GSEABulk", DiffExprsBulk - output$DiffExprsBulk <- downloadHandler( - filename = function() { - paste0("Counts and differential expression table.xlsx") - }, - content = function(file){ - file.copy("www/DESeq2_table/output_table.xlsx", - file, - copy.mode = F, - copy.date = F - ) - }, - contentType = "xlsx") - output$GSEABulk <- downloadHandler( - filename = function() { - paste0("GSEA table.xlsx") - }, - content = function(file){ - file.copy("www/GSEA_table/output_gage.xlsx", - file, - copy.mode = F, - copy.date = F - ) - }, - contentType = "xlsx") - - - #dynamic tutorials for rintrojs - { - steps <- reactive(data.frame(element = c("#Group1 + .selectize-control", - "#Group1Selector + .selectize-control", - "#Group1Add", - "#Group1Clear", - "#Group2 + .selectize-control", - "#AddOps", - "#TPCFilt + .selectize-control", - "#log2FCFilt", - "#AdjpFilt", - "#GenContrasts", - #"#res_table", - #"#DownloadTableAll", - #"#DownloadTableFilt", - #"#DownloadTableSelected", - #"#ResetTable", - "#mydropdown"), - intro = c(paste0("Start by selecting the ", - "tissue(s) that need to be ", - "contrasted. Selected ", - "tissue(s) can be removed ", - "by clicking on the name ", - "and pressing the 'backspace' ", - "or 'delete' key."), - paste0("Groups of tissues can be ", - "selected in bulk from ", - "here..."), - paste0("...and added using this ", - "button."), - paste0("The whole selection can ", - "be cleared by clicking ", - "this button."), - paste0("Tissue(s) to be contrasted ", - "against can be selected ", - "here. Note that tissues ", - "already selected in the ", - "first group are not ", - "available here and vice ", - "versa."), - paste0("This checkbox shows and ", - "hides additional options ", - "for the analysis."), - paste0("Transcripts per cell ", - "filtering filters out low ", - "expression genes from the ", - "results and plots. The ", - "filtering criteria is at ", - "least the selected amount ", - "of counts per cell in at ", - "least one Treg tissue."), - paste0("The minimum absolute log2 ", - "fold change for a gene to ", - "be considered ", - "differentially expressed."), - paste0("The maximum adjusted p.", - "value for a gene to be ", - "considered differentially ", - "expressed."), - paste0("Clicking this button will ", - "generate the contrasts ", - "between the groups ", - "selected above. This ", - "button remains disabled ", - "till both groups have at ", - "least one tissue selected."), - #paste0(), - paste0("This button can be ", - "clicked to show a summary ", - "of the instructions ", - "and features.")))) - - observeEvent(input$TutorialButton,{ - updateCheckboxInput(session = session, - inputId = "AddOps", - value = TRUE) - - introjs(session = session , options = list(steps=steps())) - }) - - steps2 <- reactive(data.frame(element = c("#res_table", - "#DownloadTableAll", - "#DownloadTableFilt", - "#DownloadTableSelected", - "#ResetTable", - "#PlotSaveOps", - "#SavePlotFormat", - "#WidthPixels", - "#HeightPixels", - "#res_volplot" - ), - intro = c(paste0("The results of the ", - "contrast are tabulated ", - "here. Column headers can ", - "be clicked to sort by ", - "that column (sorting can ", - "be done on multiple ", - "columns by pressing the ", - "'shift' key and clicking ", - "on the other header). ", - "Columns can be searched ", - "or filtered by clicking ", - "on the blank fields ", - "under the column headers.", - " Clicking a row will ", - "select the row in the ", - "table and highlight the ", - "respective gene in the ", - "volcano plot (upto 50 ", - "rows can be selected by ", - "pressing the 'shift' or ", - "'control' key)."), - paste0("Download the complete ", - "contrast results as a ", - "table."), - paste0("Download only the rows ", - "which are left after ", - "filtering (empty table if", - " no filtering done)."), - paste0("Download only the rows ", - "which have been manually ", - "selected (empty table if", - " no selection done)."), - paste0("This button resets any ", - "rows that have been ", - "selected."), - paste0("This checkbox shows and ", - "hides options related ", - "to exporting the plots."), - paste0("Select the format for ", - "exporting the plot."), - paste0("Specify the width of the ", - "plot in pixels for export."), - paste0("Specify the height of the ", - "plot in pixels for export."), - paste0("The interactive volcano ", - "plot of the contrast. ", - "Hovering over a data ", - "point would show a ", - "tooltip with additional ", - "information. Clicking on ", - "a data point will select ", - "the respective gene in ", - "the table and will also ", - "generate a column plot to ", - "visualise the expression ", - "of the selected gene in ", - "tissues. The plot can be ", - "zoomed-in by clicking and ", - "dragging a box over the ", - "area to be zoomed. ", - "Additionally the axes of ", - "the plot can be adjusted ", - "by dragging the edges of ", - "the respective axes. The ", - "zoom and/or axes can be ", - "reset by double-clicking ", - "in the empty space in the ", - "plot area or by clicking ", - "the house icon in the ", - "top-right of the plot. ", - "The plot can be exported ", - "using the export options ", - "by clicking the camera ", - "icon in the top-right of ", - "the plot. The same ", - "functionality except for ", - "data point selection also ", - "exists for the column ", - "plot.") - ))) - - observeEvent(input$TutorialButton2,{ - updateCheckboxInput(session = session, - inputId = "PlotSaveOps", - value = TRUE) - introjs(session = session , options = list(steps=steps2())) - }) - } - - } # closing bracket for server call -} - -# Create Shiny app ---- -shinyApp(ui = ui, server = server) - - - - - - + + + +#importing libraries & functions, reading in data, and setting up global options +{ + #setting global options + options(scipen = 5, digits = 4) + + #libraries + { + library(ggExtra) + library(shiny) + library(DESeq2) + library(shinyjs) + #library(shinyBS) + library(DT) + library(ggplot2) + library(ggrepel) + library(viridis) + library(plotly) + library(stringr) + library(htmlwidgets) + library(shinyWidgets) + library(shinycssloaders) + library(png) + library(htmltools) + #library(shinythemes) + #library(shinydashboard) + #library(dashboardthemes) + library(rintrojs) + library(shinyalert) + + } + + # #functions and scripts + # source("customFunctions.R") + + #data + { + #deseq2 object + DESeq_object <- readRDS(file = "data/DESeq_Object.rds") + + #formatting names for available contrasts + results_names <- resultsNames(DESeq_object) + #print(results_names) + + #aesthetic formatting of name labels + { + names(results_names) <- gsub(pattern = "tissue", + replacement = "", + x = results_names, + ignore.case = T) + + names(results_names) <- gsub(pattern = "\\.cellTypeNT", + replacement = "_Tconv", + x = names(results_names), + ignore.case = T) + + names(results_names) <- gsub(pattern = "\\.cellTypeT", + replacement = "_Tregs", + x = names(results_names), + ignore.case = T) + + # names(results_names) <- gsub(pattern = "_NT$", + # replacement = "_CD4", + # x = names(results_names), + # ignore.case = T) + # + # names(results_names) <- gsub(pattern = "_T$", + # replacement = "_Treg", + # x = names(results_names), + # ignore.case = T) + + names(results_names) <- gsub(pattern = "lymphnode", + replacement = "LN", + x = names(results_names), + ignore.case = T) + + names(results_names) <- gsub(pattern = "intraepitheliallayer", + replacement = "IEL", + x = names(results_names), + ignore.case = T) + + names(results_names) <- gsub(pattern = "laminaproprialymphocyte", + replacement = "LPL", + x = names(results_names), + ignore.case = T) + + names(results_names) <- gsub(pattern = "peyerspatches", + replacement = "PP", + x = names(results_names), + ignore.case = T) + + names(results_names) <- gsub(pattern = "_", + replacement = " ", + x = names(results_names), + ignore.case = T) + } + + + #meta data + MGI_info <- read.delim(file = "data/combined_MGI_info.txt", + sep = "\t", + stringsAsFactors = F) + + #counts + mean_norm_counts <- readRDS(file = "data/Mean_Normalised_Counts.rds") + mean_norm_counts$Gene.stable.ID <- gsub(pattern = "\\.[0-9]+$", + replacement = "", + x = rownames(mean_norm_counts), + ignore.case = T) + #mean_norm_counts2 <- readRDS(file = "Mean_Normalised_Counts.rds") + #aesthetic formatting of mean_norm_counts + { + colnames(mean_norm_counts) <- gsub(pattern = "_NT$", + replacement = "_Tconv", + x = colnames(mean_norm_counts), + ignore.case = T) + + colnames(mean_norm_counts) <- gsub(pattern = "_T$", + replacement = "_Treg", + x = colnames(mean_norm_counts), + ignore.case = T) + + colnames(mean_norm_counts) <- gsub(pattern = "lymphnode", + replacement = "LN", + x = colnames(mean_norm_counts), + ignore.case = T) + + colnames(mean_norm_counts) <- gsub(pattern = "intraepitheliallayer", + replacement = "IEL", + x = colnames(mean_norm_counts), + ignore.case = T) + + colnames(mean_norm_counts) <- gsub(pattern = "laminaproprialymphocyte", + replacement = "LPL", + x = colnames(mean_norm_counts), + ignore.case = T) + + colnames(mean_norm_counts) <- gsub(pattern = "peyerspatches", + replacement = "PP", + x = colnames(mean_norm_counts), + ignore.case = T) + } + + norm_counts <- DESeq2::counts(object = DESeq_object, normalized = T) + #aesthetic formatting of norm_counts + { + colnames(norm_counts) <- gsub(pattern = "_NT", + replacement = "_Tconv", + x = colnames(norm_counts), + ignore.case = T) + + colnames(norm_counts) <- gsub(pattern = "_T", + replacement = "_Treg", + x = colnames(norm_counts), + ignore.case = T) + + colnames(norm_counts) <- gsub(pattern = "lymphnode", + replacement = "LN", + x = colnames(norm_counts), + ignore.case = T) + + colnames(norm_counts) <- gsub(pattern = "intraepitheliallayer", + replacement = "IEL", + x = colnames(norm_counts), + ignore.case = T) + + colnames(norm_counts) <- gsub(pattern = "laminaproprialymphocyte", + replacement = "LPL", + x = colnames(norm_counts), + ignore.case = T) + + colnames(norm_counts) <- gsub(pattern = "peyerspatches", + replacement = "PP", + x = colnames(norm_counts), + ignore.case = T) + } + + #PCA, tSNE and UMAP plots + PCA_means_plots <- readRDS(file = "data/PCA_plots_means.rds") + PCA_plots <- readRDS(file = "data/PCA_plots.rds") + tSNE_plots <- readRDS(file = "data/tSNE_plots.rds") + UMAP_plots <- readRDS(file = "data/UMAP_plots.rds") + + #importing min. transcript per cell gene list object + filtered_genes_lists <- readRDS(file = "data/Filtered_Gene_Lists.rds") + + #importing TCR plots + TCR_plots_list <- list.files(path = "www/TCR_plots/", pattern = ".png") + #print(TCR_plots_list) + + ## aging_tsne_plots_list ---- + aging_tsne_plots_list <- list.files(path = "www/Aging_tsne_plots/", pattern = ".png") + aging_tsne_tissues <- gsub(pattern = " .*", replacement = "", x = aging_tsne_plots_list, ignore.case = T) + aging_tsne_tissues <- unique(gsub(pattern = "tsne_plot_", replacement = "", x = aging_tsne_tissues, ignore.case = T)) + + #importing Pathview plots + { + Pathview_plots_list <- list.files(path = "www/Pathview_plots", + pattern = ".multi\\.png") + #print(Pathview_plots_list) + + #mapping Pathview plot files to pathway names + Pathview_mapping <- read.delim(file = "data/output_gage.tab", + sep = "\t", stringsAsFactors = F)[-1,1:2] + Pathview_mapping <- + Pathview_mapping[Pathview_mapping[,1] %in% + gsub(pattern = "\\..*", + replacement = "", + x = Pathview_plots_list, + ignore.case = T),] + Pathview_mapping <- Pathview_mapping[order(Pathview_mapping[,1], + decreasing = F),] + + } + + } + + #importing functions + source("R/customFunctions.R") + +} + + +#shiny app ui +{ + # Define UI ---- + ui <- fluidPage( + + # using shinyjs to enable or disable certain sections/elements of the app + shinyjs::useShinyjs(), + # using shinyalert to enable popup messages + shinyalert::useShinyalert(), + + # Step-by-step tutorial + introjsUI(), + + #importing shinydashboard css + # tags$head( + # includeCSS(file.path('www', path = "AdminLTE.css")), + # includeCSS(file.path('www', path = "shinydashboard.css") + # )), + + #shinythemes::themeSelector(), + #theme = shinytheme("cerulean"), + #shinyDashboardThemes(theme = "grey_light"), + #shinyDashboardThemes(theme = "grey_dark"), + + #dashboardBody(), + + # App title ---- + #titlePanel("Expression Viewer"), + titlePanel("The Tissue Tregs Project"), + + tabsetPanel( + + #Landing Page panel + { + tabPanel(title = "The Study", value = "TheStudy", + + h3("General description/information of the study/data"), + + p(paste0("Some description of the study, as well as where to ", + "find additional sources such as papers and data.")), + p(paste0("Ask Adrian and Oliver as they would be able to ", + "provide a much better description")), + + hr(), + + column(width = 9), + + column(width = 3, + # style = paste0('margin-bottom:30px;border:1px solid; ', + # 'padding: 10px;'), + + wellPanel( + h4("Supplementary material for download"), + + h5("Bulk RNA-seq analysis"), + + withTags( + ul( + li(downloadLink( + outputId = "MultiQCBulk", + label = "QC and pre-processing report" + )), + li(downloadLink( + outputId = "GlobalBulk", + label = "PCA, t-SNE and UMAP plots" + )), + li(downloadLink( + outputId = "DiffExprsBulk", + label = paste0("Counts and differential ", + "expression results") + )), + li(downloadLink( + outputId = "GSEABulk", + label = "Gene set enrichment results" + )) + ) + ), + + # downloadLink( + # outputId = "MultiQCBulk", + # label = "QC and pre-processing report" + # ) + + ) + ), + + br(), + br(), + br(), + br(), + br(), + + fluidRow( + h6(paste0("Copyright 2020, Liston Lab, Babraham Institute. ", + "Babraham internal/intranet release only. ", + "Copyright licence will most likely change by the ", + "first public release as per the requirements of ", + "the publication journal and BI.")) + ) + ) + }, + + #Global Analysis panel + { + tabPanel(title = "Global Analysis", value = "GlobalAnalysis", + + h3("Exploratory analysis of the expression data"), + + p(paste0("PCA, t-SNE and UMAP analysis and plots of the ", + "RNA-seq expression data from the study.")), + + p(paste0("Plots can be downloaded by right clicking the ", + "plot and clicking on 'Save image as...'")), + + #br(), + hr(), + + h4("Bulk RNA-seq data"), + + fluidRow( + + column(width = 12, + + selectizeInput(inputId = "TriplePlot", + label = "Select Plot Type", + # choices = as.list(results_names), + #choices = c("", "PCA", "t-SNE", "UMAP"), + choices = c("PCA - tissue means", + "PCA", "t-SNE", "UMAP"), + selected = "PCA - tissue means", + #selectize = T, + multiple = F, + options = list( + placeholder = "Click to select") + ), + ) + ), + + fluidRow( + + column(width = 4, + plotOutput(outputId = "PCAplot1") %>% + withSpinner(type = 3, size = 0.5, + color.background = "#FFFFFF") + ), + column(width = 4, + plotOutput(outputId = "PCAplot2") %>% + withSpinner(type = 3, size = 0.5, + color.background = "#FFFFFF") + ), + column(width = 4, + plotOutput(outputId = "PCAplot3") %>% + withSpinner(type = 3, size = 0.5, + color.background = "#FFFFFF") + ) + ), + + hr(), + + h4("Single cell RNA-seq data"), + + p("Will be available soon") + + ) + }, + + #Bulk RNA-seq Expression panel + { + tabPanel(title = "Bulk RNA-seq Expression", + value = "BulkDiffExprs", + + fluidRow( + + column(width = 7, + h3("Generate contrasts"), + p(paste0("The differential expression analysis is performed ", + "using DESeq2. For a quick walkthrough on doing the ", + "the analysis, please click the yellow button with ", + "the question mark.")) + ), + + column(width = 5, + br(), + br(), + # actionButton(inputId = "TutorialButton", + # label = "Start tutorial"), + actionBttn(inputId = "TutorialButton", + label = "Start tutorial", + style = "material-circle", + color = "warning", + icon = icon("question")), + ) + ), + + hr(), + + fluidRow( + + column(width = 2, + + wellPanel( + + fluidRow( + + column(width = 9, + h4("Available Contrasts"), + ), + + column(width = 3, + + dropdownButton(inputId = "mydropdown", + circle = TRUE, + #status = "info", + status = "secondary", + #icon = icon("question"), + icon = icon("info"), + right = F, up = F, + width = "500px", + tooltip = tooltipOptions( + title = "Instructions"), + + h4("To Begin:"), + + h5(paste0("Start by ", + "selecting tissues", + " for the contrast", + " groups in this ", + "panel. This will", + " enable the ", + "'Generate ", + "Contrasts' button", + " at the bottom. ", + "A tissue cannot ", + "be in both ", + "groups ", + "simultaneously.")), + + h5(paste0("The drop down ", + "list and 'Add' ", + "button below ", + "each of the two ", + "selections can ", + "be used to add ", + "relevant tissues", + ", or to ", + "clear the current", + " selection. The ", + "buttons will not ", + "duplicate ", + "tissues that are ", + "already selected." + )), + + h5(paste0("Clicking the ", + "'Generate ", + "Contrasts' button", + " will perform ", + "the differential ", + "expression ", + "analysis between ", + "the two contrast ", + "groups, creating ", + "a table of ", + "results and a ", + "volcano plot on ", + "right." + )), + + h5(paste0("Clicking the ", + "'Show Advanced Options' ", + "checkbox reveals three ", + "additional options to ", + "adjust the analysis and ", + "visualisation. The ", + "'Minimum Transcripts per ", + "Cell' drop down list ", + "gives options to filter ", + "results to remove low ", + "expression genes if ", + "needed. The 'log2 Fold ", + "Change' and 'Adjusted ", + "p.value' fields are the ", + "significance criteria ", + "used in the volcano plot ", + "and can be adjusted as ", + "needed." + )), + + hr(), + h4("Interactivity:"), + + h5(paste0("The table and ", + "volcano plot are ", + "interactive. Up ", + "to 50 points/rows", + " can be selected ", + "to mark them in ", + "the volcano plot ", + "and to generate ", + "a plot of ", + "normalised counts", + " of the selected ", + "genes across all ", + "samples.")), + + h5(paste0("Alternatively, ", + "points can also", + " be selected by ", + "clicking on them", + " in the volcano", + " plot to mark ", + "them in the plot", + " and also select", + " the respective", + " rows in the ", + "table. Clicking", + " a selected ", + "point again will", + " deselect it in ", + "the plot and ", + "table.")), + + h5(paste0("The selection of ", + "points/rows can ", + "be reset at any ", + "time using the ", + "'Reset Row ", + "Selection' button", + " above the table." + )), + + h5(paste0("The plots can be ", + "zoomed in by ", + "dragging a ", + "selection box ", + "over the relevant", + " area. Reset the ", + "zoom by either ", + "double clicking ", + "anywhere in the ", + "plot or by ", + "clicking the home", + " icon on the top ", + "right of the ", + "plot.")), + + h5(paste0("The table can be ", + "filtered through ", + "the fields under ", + "the column ", + "headers. These ", + "can also be used ", + "to search the ", + "column of ", + "interest.")), + + h5(paste0("Additionaly, the ", + "table can be ", + "sorted by ", + "multiple columns ", + "by shift clicking", + " the column ", + "headers.")), + + hr(), + h4("Download/Export:"), + + h5(paste0("The results table", + " can be ", + "downloaded as ", + "tab delimited ", + "files from the ", + "three download ", + "buttons above the", + " table.")), + + h5(paste0("The plots can be ", + "downloaded via ", + "the camera icon ", + "on the top right ", + "of each plot.", + "Downloading a ", + "plot while zoomed", + " in will download", + " the zoomed in ", + "view.")), + + h5(paste0("The 'Show Plot ", + "Export Options' ", + "that becomes ", + "available above ", + "the volcano plots", + " reveals options ", + "to modify the ", + "export type and ", + "size of the ", + "plot. The ", + "format of the ", + "image (SVG/PNG/", + "JPEG) can be ", + "selected from ", + "the radio button ", + "that becomes ", + "available below ", + "the 'Generate ", + "Conrasts' button.", + " The size of the ", + "plot can be ", + "adjusted by ", + "specifying the ", + "height and width ", + "in pixels." + )) + ) + ), + ), + #h4("Available Contrasts"), + + selectizeInput(inputId = "Group1", + label = "Contrast Group 1", + # choices = as.list(results_names), + choices = list( + Tregs = results_names[ + grepl(pattern = "cellTypeT", + #grepl(pattern = "_Treg", + x = results_names, + ignore.case = T)], + Tconv = results_names[ + grepl(pattern = "cellTypeNT", + #grepl(pattern = "_Tconv", + x = results_names, + ignore.case = T)] + ), + selected = NULL, + #selectize = T, + multiple = T, + options = list( + placeholder = paste0("Click here ", + "to manually", + " select ", + "...") + ) + #) + ), + + #fluidRow( + #h5("Select available:"), + #h5("Quick add available:"), + # actionButton(inputId = "Group1SelectTconv", + # label = "Tconv"), + # actionButton(inputId = "Group1SelectTregs", + # label = "Tregs"), + selectizeInput(inputId = "Group1Selector", + label = "Add available:", + choice = list( + Treg = c("All Tregs", + "Blood Tregs", + "Lymphoid Tregs", + "Non-lymphoid Tregs", + "Gut-associated Tregs"), + Tconv = c("All Tconv", + "Blood Tconv", + "Lymphoid Tconv", + "Non-lymphoid Tconv", + "Gut-associated Tconv") + ), + selected = "All Tregs", + multiple = F, + options = list( + placeholder = "Click to select") + ), + actionButton(inputId = "Group1Add", + label = "Add"), + actionButton(inputId = "Group1Clear", + label = "Clear Selection"), + #), + + #br(), + #br(), + hr(), + #br(), + + #output of group 1 selection for debugging -- remove/ + # hide when deploying + #verbatimTextOutput(outputId = "value_Group1"), + + #p("compared against"), + + #selection for group 2 (denominator) of the contrasts + #initially disabled to allow + #disabled( + selectizeInput(inputId = "Group2", + label = "Contrast Group 2", + choices = list( + Tregs = results_names[ + grepl(pattern = "cellTypeT", + x = results_names, + ignore.case = T)], + Tconv = results_names[ + grepl(pattern = "cellTypeNT", + x = results_names, + ignore.case = T)] + ), + selected = NULL, + #selectize = T, + multiple = T, + options = list( + placeholder = paste0("Click here ", + "to manually", + " select ", + "...") + ) + ), + + #fluidRow( + #h5("Select available:"), + #h5("Quick add available:"), + # actionButton(inputId = "Group2SelectTconv", + # label = "Tconv"), + # actionButton(inputId = "Group2SelectTregs", + # label = "Tregs"), + selectizeInput(inputId = "Group2Selector", + label = "Add available:", + choice = list( + Treg = c("All Tregs", + "Blood Tregs", + "Lymphoid Tregs", + "Non-lymphoid Tregs", + "Gut-associated Tregs"), + Tconv = c("All Tconv", + "Blood Tconv", + "Lymphoid Tconv", + "Non-lymphoid Tconv", + "Gut-associated Tconv") + ), + selected = "All Tconv", + multiple = F, + options = list( + placeholder = "Click to select") + ), + actionButton(inputId = "Group2Add", + label = "Add"), + actionButton(inputId = "Group2Clear", + label = "Clear Selection"), + #), + + hr(), + + checkboxInput(inputId = "AddOps", + label = "Show Additional Options"), + includeScript(path = "js/validate-number.js"), + conditionalPanel( + condition = "input.AddOps", + selectizeInput(inputId = "TPCFilt", + label = "Minimum Transcripts per Cell", + choice = c("No filter", "0.01", + "0.1", "1"), + selected = "0.01", + multiple = F), + h5("Volcano Plot Marking Criteria"), + numericInput(inputId = "log2FCFilt", + label = HTML(paste0("log2 Fold ", + "Change (+/-)", + "
(Range: ", + "0 - 20)")), + value = 2, + min = 0, + max = 20, + step = 0.1), + numericInput(inputId = "AdjpFilt", + label = "Adjusted p.value", + value = 0.01, + min = 0, + max = 1, + step = 0.01), + ), + hr(), + #output of group 2 selection for debugging -- remove/ + # hide when deploying + #verbatimTextOutput(outputId = "value_Group2"), + + #action button for starting the DESeq2 results() + # function for generating contrasts + actionButton(inputId = "GenContrasts", + label = "Generate Contrasts" + ), + + ), + + # verbatimTextOutput(outputId = "res_error"), + # + # h6("Debug console-like thingy"), + # verbatimTextOutput(outputId = "debug_console") + + ), + + column(width = 4, + + conditionalPanel( + condition = "output.show_panels", + + #h5(HTML("
")), + br(), + fluidRow( + column(width = 10, + p(paste0("For an overview of the results, ", + "features and interactability, please ", + "click the yellow button to the right.")) + ), + column(width = 2, + # actionButton(inputId = "TutorialButton2", + # label = "Start tutorial"), + actionBttn(inputId = "TutorialButton2", + label = "Start tutorial", + style = "material-circle", + color = "warning", + icon = icon("question")), + ) + ), + + hr(), + + checkboxInput(inputId = "PlotSaveOps", + label = "Show Plot Export Options"), + + conditionalPanel( + condition = "input.PlotSaveOps", + + fluidRow( + column(width = 3,#) + radioGroupButtons(inputId = "SavePlotFormat", + label = HTML(paste0("Plot", + " Export ", + "Format")), + choices = c("SVG", "PNG", "JPEG"), + #SVG, PNG, JPEG and WEBP + #np native PDF support + selected = "SVG", + direction = "vertical", #"horizontal", + checkIcon = list( + # yes = icon("ok", + # lib = "glyphicon") + yes = icon(name = "file-image", + lib = "font-awesome") + ), + size = "normal" # xs/sm/normal/lg + ), + ), + column(width = 7,#) + includeScript(path = "js/validate-number.js"), + + numericInputIcon(inputId = "WidthPixels", + label = HTML(paste0("Plot ", + "Size in ", + " Pixels ", #"
", + "(Range: 100 ", + "- 10,000) ", + #"

", + "
", + "Width")), #in Pixels", + value = 880, + min = 100, + max = 10000, + step = 10, + width = "85%", + icon = list( + icon(name = "ruler-horizontal", + lib = "font-awesome"), + "px") + ), + + numericInputIcon(inputId = "HeightPixels", + label = "Height", #in Pixels", + value = 562, + min = 100, + max = 10000, + step = 10, + width = "85%", + icon = list( + icon(name = "ruler-vertical", + lib = "font-awesome"), + "px") + ) + ) + ) + ), + + hr(), + + h4("Differential Expression Volcano Plot"), + # h5(paste0("Click on data points to select/deselect ", + # "them in the table and show/hide their ", + # "normalised counts.")), + + plotlyOutput(outputId = "res_volplot") %>% + withSpinner(type = 3, size = 0.5, + color.background = "#FFFFFF"), + ), + + conditionalPanel( + condition = paste0( + "typeof input.res_table_rows_selected !== ", + "'undefined' && ", + "input.res_table_rows_selected.length > 0" + ), + hr(), + h4("Normalised Expression Plot"), + + plotlyOutput(outputId = "res_colplot") %>% + withSpinner(type = 3, size = 0.5, + color.background = "#FFFFFF") + ) + + ), + + column(width = 5, + + fluidRow( + conditionalPanel( + condition = "output.show_panels", + + #fluidRow( + #column(width = 11, + h4(HTML("
Results Table")), + h5("Download tab delimited file:"), + downloadButton( + outputId = "DownloadTableAll", + label = "Complete Results" + ), + downloadButton( + outputId = "DownloadTableFilt", + label = "Filtered Table" + ), + downloadButton( + outputId = "DownloadTableSelected", + label = "Selected Rows Only" + ) + #) + #), + ), + ), + #br(), + conditionalPanel( + condition = "output.show_panels", + fluidRow( + column(width = 12, + hr(), + #h5("Row Selection Options:"), + actionButton(inputId = "ResetTable", + label = "Reset Row Selection" + ) + ) + ), + br(), + ), + + # conditionalPanel( + # condition = paste0( + # "typeof output.res_table !== 'undefined'" + # ), + #shinyjs::hidden( + fluidRow( + #shinyjs::hidden( + DT::dataTableOutput(outputId = "res_table") #%>% + # withSpinner(type = 3, size = 0.5, + # color.background = "#FFFFFF") + #) + ) + ) + ) + ) + }, + + #Pathway analysis panel + { + tabPanel(title = "Pathway Analysis", value = "PathwayAnalysis", + + h3("Visualise enriched KEGG pathways"), + + p(paste0("The results of the pathway analsis using GAGE and ", + "Pathview can be visualised below.")), + + hr(), + + #vr(), + + column(width = 9, + + fluidRow( + + #h4("Visualise Enriched KEGG Pathways"), + + column(width = 6, + #br(), + br(), + + selectizeInput(inputId = "PathviewPlotSelect", + label = "Select KEGG Pathway", + # choices = as.list(results_names), + #choices = c("", "PCA", "t-SNE", "UMAP"), + choices = paste(Pathview_mapping[,1], + Pathview_mapping[,2], + sep = " - "), + selected = paste(Pathview_mapping[,1], + Pathview_mapping[,2], + sep = " - ")[1], + #selectize = T, + multiple = F, + options = list( + placeholder = "Click to select"), + width = "600px" + ), + + downloadButton( + outputId = "DownloadPathviewSelected", + label = "Selected Pathway" + ), + + downloadButton( + outputId = "DownloadPathviewAll", + label = "All Pathways" + ) + + ), + + column(width = 3, + + imageOutput(outputId = "PathviewPlotLegend", + width = "100%", + height = "100%") %>% + withSpinner(type = 3, size = 0.5, + color.background = "#FFFFFF"), + br() + ), + + column(width = 3, + + br(), + imageOutput(outputId = "PathviewPlotLegend2", + width = "100%", + height = "100%") %>% + withSpinner(type = 3, size = 0.5, + color.background = "#FFFFFF") + )#, + ), + + imageOutput(outputId = "PathviewPlotImage", + width = 1342*0.35, + height = 1451*0.35) %>% + withSpinner(type = 3, size = 0.5, + color.background = "#FFFFFF") + + ) + ) + + }, + + #Single Cell Expression panel + # { + # tabPanel(title = "Single Cell Expression", value = "SingleCellExprs", + # + # ) + # }, + + #TCR panel ---- + { + tabPanel(title = "TCR", value = "TCR", + + h3("TCR repertoire analysis"), + + p(paste0("Description of the TCR repertoire analysis - ", + "ask Orian")), + + #br(), + hr(), + + fluidRow( + + column(width = 4, + + selectizeInput(inputId = "TCRChordSelect", + label = "Select Tissue", + # choices = as.list(results_names), + #choices = c("", "PCA", "t-SNE", "UMAP"), + choices = gsub(pattern = "_.*", + replacement = "", + x = TCR_plots_list, + ignore.case = T), + selected = "Blood", + #selectize = T, + multiple = F, + options = list( + placeholder = "Click to select") + ), + + downloadButton( + outputId = "DownloadTCRSelected", + label = "Selected Tissue" + ), + + downloadButton( + outputId = "DownloadTCRAll", + label = "All Tissues" + ) + + ), + + column(width = 8, + + imageOutput(outputId = "TCRChordImage", + width = 4344*0.15, + height = 4601*0.15) %>% + withSpinner(type = 3, size = 0.5, + color.background = "#FFFFFF") + + ) + + ) + ) + }, + # Aging t-SNE panel ---- + # It might be worth putting the pngs together into one plot + { + tabPanel( + title = "Aging t-SNE", + value = "AgingTSNE", + h3("t-SNE plots across 6 time-points"), + p(paste0("Further description?")), + hr(), + fluidRow( + column( + width = 3, + selectizeInput( + inputId = "agingTSNEtissue", + label = "Select Tissue", + choices = aging_tsne_tissues, + #selectize = T, + multiple = FALSE, + options = list( + placeholder = "Click to select") + ), + downloadButton( + outputId = "DownloadAgingTSNEtissue", + label = "Selected Tissue" + ), + downloadButton( + outputId = "DownloadAgingTSNEAll", + label = "All Tissues" + ), + actionButton("browser", "browser") + ), + column( + width = 4, + imageOutput( + outputId = "agingTSNEImage1", + width = 4344*0.15, + height = 4601*0.15 + ) %>% + withSpinner(type = 3, size = 0.5, color.background = "#FFFFFF") + ), + column( + width = 4, + imageOutput( + outputId = "agingTSNEImage2", + width = 4344*0.15, + height = 4601*0.15 + ) %>% + withSpinner(type = 3, size = 0.5, color.background = "#FFFFFF") + ) + ) + ) + } + ), + ) + ## ui fluid page ends at bracket immediately above +} + + +#shiny app server +{ + + # Define server logic ---- + server <- function(input, output, session) { + + #reading inputs for debugging + #output$value_Group1 <- renderPrint({ (input$Group1) }) + #output$value_Group2 <- renderPrint({ (input$Group2) }) + + #output$value_padj <- renderPrint({ (input$Padj_cutoff) }) + + #debug console output + # output$debug_console <- renderPrint(results_names[ + # grepl(pattern = "_NT", + # x = results_names, + # ignore.case = T)]) + # output$debug_console <- renderPrint(class(input$Group1)) + + #external variable handles so that they do not get destroyed once relevant + #gui updates and rendering are performed + global_data <- reactiveValues(result = NULL, + base_traces = 0, + #plotly traces are 0 indexed so count is ct-1 + trace_count = 0, + #trace_count = base_traces-1, + dt = NULL, + #volplot_selected = NULL#, + #volplot = NULL, + #colplot = NULL + left_log = 0, + right_log = 0, + first_run_done = F + ) + + #live updating UI elements + observe({ + + updateSelectInput(session = session, + inputId = "Group1", + label = "Contrast Group 1", + # choices = as.list(results_names[ + # !(results_names %in% input$Group2) + # ]), + choices = list( + Tregs = results_names[ + grepl(pattern = "cellTypeT", + x = results_names, + ignore.case = T) & !(results_names %in% + input$Group2)], + Tconv = results_names[ + grepl(pattern = "cellTypeNT", + x = results_names, + ignore.case = T) & !(results_names %in% + input$Group2)] + ), + selected = input$Group1) + + updateSelectInput(session = session, + inputId = "Group2", + label = "Contrast Group 2", + # choices = as.list(results_names[ + # !(results_names %in% input$Group1) + # ]), + choices = list( + Tregs = results_names[ + grepl(pattern = "cellTypeT", + x = results_names, + ignore.case = T) & !(results_names %in% + input$Group1)], + Tconv = results_names[ + grepl(pattern = "cellTypeNT", + x = results_names, + ignore.case = T) & !(results_names %in% + input$Group1)] + ), + selected = input$Group2) + + updateNumericInput(session = session, + inputId = "HeightPixels", + # label = tags$div(HTML(paste0(' Plot ', + # 'Height (pixels)') + # ) + # ), + value = ifelse(!is.numeric(input$HeightPixels), 562, + ifelse(input$HeightPixels > 10000, + 10000, ifelse( + input$HeightPixels < 100, 100, + round(input$HeightPixels)) + ) + ), + min = 100, + max = 10000, + step = 10 + ) + + updateNumericInput(session = session, + inputId = "WidthPixels", + # label = tags$div(HTML(paste0(' Plot ', + # 'Width (pixels)') + # ) + # ), + value = ifelse(!is.numeric(input$WidthPixels), 880, + ifelse(input$WidthPixels > 10000, + 10000, ifelse( + input$WidthPixels < 100, 100, + round(input$WidthPixels)) + ) + ), + min = 100, + max = 10000, + step = 10 + ) + + updateNumericInput(session = session, + inputId = "log2FCFilt", + value = ifelse(input$log2FCFilt > 20, 20, + ifelse(input$log2FCFilt < 0, + -(input$log2FCFilt), + ifelse( + !is.numeric(input$log2FCFilt) | + is.na(input$log2FCFilt) | + is.null(input$log2FCFilt) | + input$log2FCFilt == "" | + input$log2FCFilt == 0, + 2, input$log2FCFilt) + ) + ), + min = 0, max = 20, step = 0.1 + ) + updateNumericInput(session = session, + inputId = "AdjpFilt", + value = ifelse(input$AdjpFilt > 1, 1, + ifelse(input$AdjpFilt < 0, + -(input$AdjpFilt), + ifelse( + !is.numeric(input$AdjpFilt) | + is.na(input$AdjpFilt) | + is.null(input$AdjpFilt) | + input$AdjpFilt == "" | + input$AdjpFilt == 0, + 0.01, input$AdjpFilt) + ) + ), + min = 0, max = 1, step = 0.01 + ) + #print(input$AdjpFilt) + + }) + + #update available samples in contrast group 1 selection buttons + { + #adding entries to selection based on group + observeEvent(input$Group1Add, { + group1_selection <- unlist(str_split(string = input$Group1Selector, + pattern = " ")) + + #print(results_names) + #print(group1_selection) + if(group1_selection[2] == "Tregs"){ + available <- results_names[grepl(pattern = "cellTypeT", + x = results_names, + ignore.case = T)] + } + else{ + available <- results_names[grepl(pattern = "cellTypeNT", + x = results_names, + ignore.case = T)] + } + #print(available) + + if(group1_selection[1] == "Blood"){ + updateSelectInput(session = session, + inputId = "Group1", + label = "Contrast Group 1", + choices = list( + Tregs = results_names[ + grepl(pattern = "cellTypeT", + x = results_names, + ignore.case = T) & !(results_names %in% + input$Group2)], + Tconv = results_names[ + grepl(pattern = "cellTypeNT", + x = results_names, + ignore.case = T) & !(results_names %in% + input$Group2)] + ), + selected = c(input$Group1, available[ + grepl(pattern = "Blood", + x = available, + ignore.case = T) & !(available %in% + input$Group2)]) + ) + } + else if(group1_selection[1] == "Lymphoid"){ + updateSelectInput(session = session, + inputId = "Group1", + label = "Contrast Group 1", + choices = list( + Tregs = results_names[ + grepl(pattern = "cellTypeT", + x = results_names, + ignore.case = T) & !(results_names %in% + input$Group2)], + Tconv = results_names[ + grepl(pattern = "cellTypeNT", + x = results_names, + ignore.case = T) & !(results_names %in% + input$Group2)] + ), + selected = c(input$Group1, available[ + grepl(pattern = "(LymphNode|Spleen|PeyersPatches)", + x = available, + ignore.case = T) & !(available %in% + input$Group2)]) + ) + } + else if(group1_selection[1] == "Non-lymphoid"){ + updateSelectInput(session = session, + inputId = "Group1", + label = "Contrast Group 1", + choices = list( + Tregs = results_names[ + grepl(pattern = "cellTypeT", + x = results_names, + ignore.case = T) & !(results_names %in% + input$Group2)], + Tconv = results_names[ + grepl(pattern = "cellTypeNT", + x = results_names, + ignore.case = T) & !(results_names %in% + input$Group2)] + ), + selected = c(input$Group1, available[ + grepl(pattern = "(Kidney|Liver|Lung|Pancreas)", + x = available, + ignore.case = T) & !(available %in% + input$Group2)]) + ) + } + else if(group1_selection[1] == "Gut-associated"){ + updateSelectInput(session = session, + inputId = "Group1", + label = "Contrast Group 1", + choices = list( + Tregs = results_names[ + grepl(pattern = "cellTypeT", + x = results_names, + ignore.case = T) & !(results_names %in% + input$Group2)], + Tconv = results_names[ + grepl(pattern = "cellTypeNT", + x = results_names, + ignore.case = T) & !(results_names %in% + input$Group2)] + ), + selected = c(input$Group1, available[ + grepl(pattern = "(IntraEpithelialLayer|LaminaPropriaLymphocyte)", + x = available, + ignore.case = T) & !(available %in% + input$Group2)]) + ) + } + else{ + updateSelectInput(session = session, + inputId = "Group1", + label = "Contrast Group 1", + choices = list( + Tregs = results_names[ + grepl(pattern = "cellTypeT", + x = results_names, + ignore.case = T) & !(results_names %in% + input$Group2)], + Tconv = results_names[ + grepl(pattern = "cellTypeNT", + x = results_names, + ignore.case = T) & !(results_names %in% + input$Group2)] + ), + selected = c(input$Group1, available[ + !(available %in% input$Group2)]) + ) + } + + }) + + #clear selection + observeEvent(input$Group1Clear, { + + updateSelectInput(session = session, + inputId = "Group1", + label = "Contrast Group 1", + choices = list( + CD4s = results_names[ + grepl(pattern = "cellTypeNT", + x = results_names, + ignore.case = T) & !(results_names %in% + input$Group2)], + Tregs = results_names[ + grepl(pattern = "cellTypeT", + x = results_names, + ignore.case = T) & !(results_names %in% + input$Group2)] + ), + selected = NULL + ) + + }) + + #old button code + { + # #all CD4s + # observeEvent(input$Group1SelectTconv, { + # + # updateSelectInput(session = session, + # inputId = "Group1", + # label = "Contrast Group 1", + # choices = list( + # CD4s = results_names[ + # grepl(pattern = "cellTypeNT", + # x = results_names, + # ignore.case = T) & !(results_names %in% + # input$Group2)], + # Tregs = results_names[ + # grepl(pattern = "cellTypeT", + # x = results_names, + # ignore.case = T) & !(results_names %in% + # input$Group2)] + # ), + # selected = c(input$Group1, results_names[ + # grepl(pattern = "cellTypeNT", + # x = results_names, + # ignore.case = T) & !(results_names %in% + # input$Group2)]) + # ) + # + # }) + # + # #all Tregs + # observeEvent(input$Group1SelectTregs, { + # + # updateSelectInput(session = session, + # inputId = "Group1", + # label = "Contrast Group 1", + # choices = list( + # CD4s = results_names[ + # grepl(pattern = "cellTypeNT", + # x = results_names, + # ignore.case = T) & !(results_names %in% + # input$Group2)], + # Tregs = results_names[ + # grepl(pattern = "cellTypeT", + # x = results_names, + # ignore.case = T) & !(results_names %in% + # input$Group2)] + # ), + # selected = c(input$Group1, results_names[ + # grepl(pattern = "cellTypeT", + # x = results_names, + # ignore.case = T) & !(results_names %in% + # input$Group2)]) + # ) + # + # }) + # + # #all remaining + # # observeEvent(input$Group1SelectAll, { + # # + # # updateSelectInput(session = session, + # # inputId = "Group1", + # # label = "Contrast Group 1", + # # choices = list( + # # CD4s = results_names[ + # # grepl(pattern = "cellTypeNT", + # # x = results_names, + # # ignore.case = T) & !(results_names %in% + # # input$Group2)], + # # Tregs = results_names[ + # # grepl(pattern = "cellTypeT", + # # x = results_names, + # # ignore.case = T) & !(results_names %in% + # # input$Group2)] + # # ), + # # selected = c(input$Group1, results_names[ + # # grepl(pattern = "cellTypeNT", + # # x = results_names, + # # ignore.case = T) & !(results_names %in% + # # input$Group2)], + # # results_names[ + # # grepl(pattern = "cellTypeT", + # # x = results_names, + # # ignore.case = T) & !(results_names %in% + # # input$Group2)]) + # # ) + # # + # # }) + # + } + + } + + #update available samples in contrast group 2 selection buttons + { + #adding entries to selection based on group + observeEvent(input$Group2Add, { + group2_selection <- unlist(str_split(string = input$Group2Selector, + pattern = " ")) + #print(group1_selection) + if(group2_selection[2] == "Tregs"){ + available <- results_names[grepl(pattern = "cellTypeT", + x = results_names, + ignore.case = T)] + } + else{ + available <- results_names[grepl(pattern = "cellTypeNT", + x = results_names, + ignore.case = T)] + } + #print(available) + + if(group2_selection[1] == "Blood"){ + updateSelectInput(session = session, + inputId = "Group2", + label = "Contrast Group 2", + choices = list( + Tregs = results_names[ + grepl(pattern = "cellTypeT", + x = results_names, + ignore.case = T) & !(results_names %in% + input$Group1)], + Tconv = results_names[ + grepl(pattern = "cellTypeNT", + x = results_names, + ignore.case = T) & !(results_names %in% + input$Group1)] + ), + selected = c(input$Group2, available[ + grepl(pattern = "Blood", + x = available, + ignore.case = T) & !(available %in% + input$Group1)]) + ) + } + else if(group2_selection[1] == "Lymphoid"){ + updateSelectInput(session = session, + inputId = "Group2", + label = "Contrast Group 2", + choices = list( + Tregs = results_names[ + grepl(pattern = "cellTypeT", + x = results_names, + ignore.case = T) & !(results_names %in% + input$Group1)], + Tconv = results_names[ + grepl(pattern = "cellTypeNT", + x = results_names, + ignore.case = T) & !(results_names %in% + input$Group1)] + ), + selected = c(input$Group2, available[ + grepl(pattern = "(LymphNode|Spleen|PeyersPatches)", + x = available, + ignore.case = T) & !(available %in% + input$Group1)]) + ) + } + else if(group2_selection[1] == "Non-lymphoid"){ + updateSelectInput(session = session, + inputId = "Group2", + label = "Contrast Group 2", + choices = list( + Tregs = results_names[ + grepl(pattern = "cellTypeT", + x = results_names, + ignore.case = T) & !(results_names %in% + input$Group1)], + Tconv = results_names[ + grepl(pattern = "cellTypeNT", + x = results_names, + ignore.case = T) & !(results_names %in% + input$Group1)] + ), + selected = c(input$Group2, available[ + grepl(pattern = "(Kidney|Liver|Lung|Pancreas)", + x = available, + ignore.case = T) & !(available %in% + input$Group1)]) + ) + } + else if(group2_selection[1] == "Gut-associated"){ + updateSelectInput(session = session, + inputId = "Group2", + label = "Contrast Group 2", + choices = list( + Tregs = results_names[ + grepl(pattern = "cellTypeT", + x = results_names, + ignore.case = T) & !(results_names %in% + input$Group1)], + Tconv = results_names[ + grepl(pattern = "cellTypeNT", + x = results_names, + ignore.case = T) & !(results_names %in% + input$Group1)] + ), + selected = c(input$Group2, available[ + grepl(pattern = "(IntraEpithelialLayer|LaminaPropriaLymphocyte)", + x = available, + ignore.case = T) & !(available %in% + input$Group1)]) + ) + } + else{ + updateSelectInput(session = session, + inputId = "Group2", + label = "Contrast Group 2", + choices = list( + Tregs = results_names[ + grepl(pattern = "cellTypeT", + x = results_names, + ignore.case = T) & !(results_names %in% + input$Group1)], + Tconv = results_names[ + grepl(pattern = "cellTypeNT", + x = results_names, + ignore.case = T) & !(results_names %in% + input$Group1)] + ), + selected = c(input$Group2, available[ + !(available %in% input$Group1)]) + ) + } + + }) + + #clear selection + observeEvent(input$Group2Clear, { + + updateSelectInput(session = session, + inputId = "Group2", + label = "Contrast Group 2", + choices = list( + CD4s = results_names[ + grepl(pattern = "cellTypeNT", + x = results_names, + ignore.case = T) & !(results_names %in% + input$Group1)], + Tregs = results_names[ + grepl(pattern = "cellTypeT", + x = results_names, + ignore.case = T) & !(results_names %in% + input$Group1)] + ), + selected = NULL + ) + + }) + + #old button code + { + # #all CD4s + # observeEvent(input$Group2SelectTconv, { + # + # updateSelectInput(session = session, + # inputId = "Group2", + # label = "Contrast Group 2", + # choices = list( + # CD4s = results_names[ + # grepl(pattern = "cellTypeNT", + # x = results_names, + # ignore.case = T) & !(results_names %in% + # input$Group1)], + # Tregs = results_names[ + # grepl(pattern = "cellTypeT", + # x = results_names, + # ignore.case = T) & !(results_names %in% + # input$Group1)] + # ), + # selected = c(input$Group2, results_names[ + # grepl(pattern = "cellTypeNT", + # x = results_names, + # ignore.case = T) & !(results_names %in% + # input$Group1)]) + # ) + # + # }) + # + # #all Tregs + # observeEvent(input$Group2SelectTregs, { + # + # updateSelectInput(session = session, + # inputId = "Group2", + # label = "Contrast Group 2", + # choices = list( + # CD4s = results_names[ + # grepl(pattern = "cellTypeNT", + # x = results_names, + # ignore.case = T) & !(results_names %in% + # input$Group1)], + # Tregs = results_names[ + # grepl(pattern = "cellTypeT", + # x = results_names, + # ignore.case = T) & !(results_names %in% + # input$Group1)] + # ), + # selected = c(input$Group2, results_names[ + # grepl(pattern = "cellTypeT", + # x = results_names, + # ignore.case = T) & !(results_names %in% + # input$Group1)]) + # ) + # + # }) + # + # #all remaining + # # observeEvent(input$Group2SelectAll, { + # # + # # updateSelectInput(session = session, + # # inputId = "Group2", + # # label = "Contrast Group 2", + # # choices = list( + # # CD4s = results_names[ + # # grepl(pattern = "cellTypeNT", + # # x = results_names, + # # ignore.case = T) & !(results_names %in% + # # input$Group1)], + # # Tregs = results_names[ + # # grepl(pattern = "cellTypeT", + # # x = results_names, + # # ignore.case = T) & !(results_names %in% + # # input$Group1)] + # # ), + # # selected = c(input$Group2, results_names[ + # # grepl(pattern = "cellTypeNT", + # # x = results_names, + # # ignore.case = T) & !(results_names %in% + # # input$Group1)], + # # results_names[ + # # grepl(pattern = "cellTypeT", + # # x = results_names, + # # ignore.case = T) & !(results_names %in% + # # input$Group1)]) + # # ) + # # + # # }) + # + } + + } + + #enabling / disabling UI elements + observe({ + shinyjs::toggleState(id = "GenContrasts", + condition = !(length(input$Group1) < 1 | + length(input$Group2) < 1) && + (is.numeric(input$log2FCFilt) & + is.numeric(input$AdjpFilt)) + ) + }) + #output$show_panels <- reactive({FALSE}) + #outputOptions(x = output, name = "show_panels", suspendWhenHidden = FALSE) + shinyjs::hide(id = "res_table") + #shinyjs::disable(id = "res_table") + + + #generating contrasts via the DESeq2 results() function + observeEvent(input$GenContrasts, { + + #splash alert for the user informing them that the analysis may take some + # time + if(!global_data$first_run_done){ + # sendSweetAlert(session = session, + # title = "Computing contrasts", + # text = paste0("The computation may take some time during ", + # "which the tool may appear to not be doing ", + # "anything. Once the results are computed ", + # "they will populate the tab."), + # type = "info") + shinyalert(title = "Computing contrasts", + text = paste0("The computation may take some time during ", + "which the tool may appear to not be doing ", + "anything. Once the results are computed, ", + "they will populate the tab."), + type = "info", + animation = F, #"slide-from-bottom", + inputId = "AlertFirstRunDone") + global_data$first_run_done <- T + } + + + #ensuring that the minimum log2FC and minimum adj.p.value fields weren't + # left blank for the fraction of the second it takes to disable the + # generate contrast button + req(input$log2FCFilt, input$AdjpFilt, input$TPCFilt) + + #disabling the GenContrasts button so that users may not spam it + #is renabled at the end of this observeEvent block + shinyjs::disable(id = "GenContrasts") + #enabling the results table rendered object + #shinyjs::show(id = "res_table") + + #resetting the previous output if it exists + output$res_error <- NULL + output$res_table <- NULL + output$res_volplot <- NULL + output$res_colplot <- NULL + + #print(input$TPCFilt) + #checking which filter list to send to the contrast + if(input$TPCFilt == "No filter"){ + tpc_filtered_list <- NULL + } + else{ + tpc_filtered_list <- filtered_genes_lists[[paste0("Filt", + input$TPCFilt)]] + } + + #performing the contrast + result <- GenerateContrasts(DESeq_object = DESeq_object, + group1 = input$Group1, + group2 = input$Group2, + meta_data = MGI_info, + mean_norm_counts = mean_norm_counts, + tpc_filtered_list = tpc_filtered_list + ) + #print(class(result[[2]])) + #global_data$result <- result + + #filtering results for only genes for which actual logFC exists + # actually bad idea because we would lose genes which are not being + # in one contrast group (e.g., those which are only expressed in Treg and + # not Tconv) + # actual_logFC <- + # + # (is.finite(log2(rowMeans(norm_counts[,2:4])) - log2(rowMeans(norm_counts[,6:8]))))[order()] + + #interpretting the type of output and updating relevant UI elements + if(length(result) == 2){ + + #outputting error + output$res_error <- renderText({ result[[1]] }) + + #saving the results in a shared variable + global_data$result <- result[[2]] + + result <- result[[2]] + + } + else if(class(result) == "data.frame"){ + #saving the results in a shared variable + global_data$result <- result + + } + else if(class(result) == "character"){ + output$res_error <- renderText({ result }) + } + + #print(summary(isolate(global_data$result))) + #calculating the fold change values for the left and right flanking + # columns of the NA/NaN/Inf/-Inf genes in the volcano plot + # --can be sent to the plotting function too, but I already have that up + # -- and running; this is for the table/data point selection and traces + global_data$left_log <- -(ceiling(max(abs( + isolate(global_data$result$DESeq2.Beta.Coefficients[ + is.finite(isolate(global_data$result$log2.Fold.Change))])))) + 5) #* -1 + #))) + #print(isolate(global_data$left_log)) + global_data$right_log <- (ceiling(max(abs( + isolate(global_data$result$DESeq2.Beta.Coefficients[ + is.finite(isolate(global_data$result$log2.Fold.Change))])))) + 5) + #))) + # print(isolate(global_data$right_log)) + + + #processing data frame to be displayed as DataTable + if(class(result) == "data.frame"){ + + #replacing zero p.values/adj.p.values with smallest IEEE supported value + global_data$result$p.value[global_data$result$p.value == 0] <- + .Machine$double.xmin + global_data$result$Adjusted.p.value[ + global_data$result$Adjusted.p.value == 0] <- .Machine$double.xmin + + result <- global_data$result + + #removing extra columns + result$Standard.Error <- NULL + result$Statistic <- NULL + result$p.value <- NULL + + #formatting the numeric columns out here rather than through DT::format* + # so that the filter ranges do not end up with 15 decimal places + result$Mean.Counts <- round(x = result$Mean.Counts, digits = 3) + result$log2.Fold.Change <- round(x = result$log2.Fold.Change, + digits = 3) + result$DESeq2.Beta.Coefficients <- + round(x = result$DESeq2.Beta.Coefficients, digits = 3) + result$Adjusted.p.value <- signif(x = result$Adjusted.p.value, + digits = 3) + + #adding selection column for sorting selected rows -- not being used atm + result$Selected <- "No" + + #outputting results + output$res_table <- DT::renderDataTable({ + DT::datatable(elementId = "res_table", + data = result, + #caption = "Differential expression results", + colnames = gsub(pattern = "\\.", + replacement = " ", + x = colnames(result), + ignore.case = T), + #extensions = "Buttons", + options = list(searching = T, + searchHighlight = TRUE, + dom = "lrtip", + #buttons = c('csv', 'excel', 'pdf'), + #order = list(list(5, 'asc')) + order = list(list(6, 'asc')) + ), + filter = list(position = "top", + plain = F + ), + editable = F, + rownames = F) #%>% + # DT::formatRound(columns = c("Mean.Counts"), + # digits = 2, + # mark = "") %>% + # DT::formatRound(columns = c("log2.Fold.Change"), + # digits = 3, + # mark = "") %>% + # DT::formatSignif(columns = c("Adjusted.p.value"), + # digits = 3, + # mark = "") + }) + + #used for updating DataTable values for the 'Selected' column + global_data$dt <- result + + } + + + #checking if a contrast was generated to create the volcano plot + if(class(result) == "data.frame"){ + + #interpreting the label names for the plot title + names_group1 <- names(results_names[results_names %in% + isolate(input$Group1)]) + names_group2 <- names(results_names[results_names %in% + isolate(input$Group2)]) + + #turning off warning to hide the persistent "Warning: 'scattergl' + # objects don't have these attributes: 'hoveron'" warnings. Comment the + # below code when debugging the plotting code + #restoreWarn <- getOption("warn") + #options(warn = -1) + + #generating the ggplotly plot + # output$res_volplot <- renderPlotly({ + # GenerateVolcanoPlot(plot_data = isolate(global_data$result), + # names_group1 = names_group1, + # names_group2 = names_group2, + # format = tolower(isolate(input$SavePlotFormat)), + # width = isolate(input$WidthPixels), + # height = isolate(input$HeightPixels) + # ) + p <- GenerateVolcanoPlot(plot_data = isolate(global_data$result), + names_group1 = names_group1, + names_group2 = names_group2, + format = tolower(isolate(input$SavePlotFormat)), + width = isolate(input$WidthPixels), + height = isolate(input$HeightPixels), + log2FCFilt = input$log2FCFilt, + AdjpFilt = input$AdjpFilt + ) + output$res_volplot <- renderPlotly({p}) + p <- plotly_build(p) + global_data$base_traces <- length(p$x$data) + global_data$trace_count <- global_data$base_traces-1 + #plotly traces are zero indexed so trace_count is base_traces-1 + + #restoring warnings + #options(warn = restoreWarn) + + } + + #renabling the disabled GenContrasts button + shinyjs::enable(id = "GenContrasts") + #shinyjs::enable(id = "res_table") + + #showing hidden ui controls and elements + shinyjs::show(id = "res_table") + output$show_panels <- reactive({TRUE}) #for multiple conditionalPanels + outputOptions(x = output, name = "show_panels", suspendWhenHidden = FALSE) + + # #update the SavePlotFormat once more to allow saving the default SVG + # updateRadioGroupButtons(session = session, + # inputId = "SavePlotFormat", + # selected = isolate(input$SavePlotFormat)) + + #resuming the DT row selection observer + #res_table_row_observer$resume() + + }) + + + #generating bar plot of normalised counts for the genes of the rows selected + # in the rendered DataTable + observeEvent(input$res_table_rows_selected, ignoreNULL = F, { + + #resetting any existing plot + output$res_colplot <- NULL + + #remove any existing plotly traces on the volcano plot + # plotly traces are zero indexed so min count is global_data$base_traces-1 + #if(global_data$trace_count > 2){ + if(global_data$trace_count > global_data$base_traces-1){ + plotlyProxy(outputId = "res_volplot", session = session) %>% + plotlyProxyInvoke("deleteTraces", + #as.list(c(2:global_data$trace_count)) + as.list(c( + global_data$base_traces:global_data$trace_count)) + ) + + global_data$trace_count <- global_data$base_traces-1 + #print(global_data$trace_count) + } + + #if there is at least one row selected + if(!is.null(input$res_table_rows_selected)){ + + #hard limiting max selectable rows to first 50 selected + if(length(input$res_table_rows_selected) > 50){ + selected_rows <- input$res_table_rows_selected[1:50] + DT::dataTableProxy("res_table") %>% selectRows(selected_rows) + } + else{ + selected_rows <- input$res_table_rows_selected + } + + selection <- isolate(global_data$result[selected_rows, 1:2]) + + #generating the counts plot + output$res_colplot <- renderPlotly({ + GenerateColPlot(mean_norm_counts = mean_norm_counts, + selection = selection, + format = tolower(isolate(input$SavePlotFormat)), + width = isolate(input$WidthPixels), + height = isolate(input$HeightPixels) + ) + }) + + #adding traces to the plotly volcano plot + #need cols 5 and 9 from selection + selection <- isolate(global_data$result[selected_rows,]) + #print(summary(selection)) + + #colnames: 1-Gene.ID 2-Gene.Symbol 3-Description 4-Mean.Counts + # 5-log2.Fold.Change 6-DESeq2.Beta.Coefficients 7-p.value + # 8-Adjusted.p.value 9-Standard.Error 10-Statistic + + #output$debug_console <- renderPrint(selection) + + #plotly does not show traces for single items -- + #duplicating item if single + if(nrow(selection) == 1){ + #checking if log2.Fold.Change exists, then use DESeq2.Beta.Coefficients + # else use max/min for the right/left columns + if(is.finite(selection[1,5])){ + selected_x <- c(selection[1,6],selection[,6]) + } + else{ + if(selection[1,6] < 0) selected_x <- + c(isolate(global_data$left_log), isolate(global_data$left_log)) + else if(selection[1,6] > 0) selected_x <- + c(isolate(global_data$right_log), isolate(global_data$right_log)) + } + #selected_y <- c(selection[1,9],selection[,9]) + selected_y <- c(selection[1,8],selection[,8]) + + #if MGI or other symbols are missing + #if(selection[,2] == ""){ + if(is.na(selection[,2]) | selection[,2] == ""){ + # selected_text <- c(str_extract(selection[,1], '\\w*')[1], + # str_extract(selection[,1], '\\w*')) + selected_text <- c(gsub(pattern = ";.*", + replacement = "", + x = selection[,1], + ignore.case = T)[1], + gsub(pattern = ";.*", + replacement = "", + x = selection[,1], + ignore.case = T) + ) + #print(selected_text) + } + else{ + # selected_text <- c(str_extract(selection[,2], '\\w*')[1], + # str_extract(selection[,2], '\\w*')) + selected_text <- c(gsub(pattern = ";.*", + replacement = "", + x = selection[,2], + ignore.case = T)[1], + gsub(pattern = ";.*", + replacement = "", + x = selection[,2], + ignore.case = T) + ) + #print(selected_text) + } + } + else{ + #selected_x <- selection[,5] + selected_x <- selection[,6] + #checking if log2.Fold.Change exists, then use DESeq2.Beta.Coefficients + # else use max/min for the right/left columns + selected_x[!is.finite(selection[,5]) & selection[,6] < 0] <- + isolate(global_data$left_log) + selected_x[!is.finite(selection[,5]) & selection[,6] > 0] <- + isolate(global_data$right_log) + + #selected_y <- selection[,9] + selected_y <- selection[,8] + + #if MGI or other symbols are missing + #if(selection[,2] == ""){ + if(any(is.na(selection[,2])) | any(selection[,2] == "")){ + #print(selection[,2]) + #selected_text <- str_extract(selection[,1], '\\w*') + selected_text <- gsub(pattern = ";.*", + replacement = "", + x = selection[,1], + ignore.case = T) + selected_text[-(which(is.na(selection[,2]) | + selection[,2] == ""))] <- + gsub(pattern = ";.*", + replacement = "", + x = selection[-(which(is.na(selection[,2]) | + selection[,2] == "")),2], + ignore.case = T) + # selected_text[-(which(is.na(selection[,2]) | + # selection[,2] == ""))] <- + # str_extract(selection[-(which(is.na(selection[,2]) | + # selection[,2] == "")),2], '\\w*') + #print(selected_text) + } + else{ + #selected_text <- str_extract(selection[,2], '\\w*') + selected_text <- gsub(pattern = ";.*", + replacement = "", + x = selection[,2], + ignore.case = T) + } + } + + plotlyProxy(outputId = "res_volplot", session = session) %>% + plotlyProxyInvoke("addTrace", + list(#x = c(selection[1,5],selection[,5]), + #y = -log10(c(selection[1,9], selection[,9])), + x = selected_x, + y = -log10(selected_y), + type = "scattergl", + name = "Selected genes", + mode = "markers+text", + marker = list(#symbol = "x", + symbol = "circle-open", + size = 10, + #colorscale = "Viridis" + color = "blue" + ), + # text = c(str_extract(selection[,2], '\\w*')[1], + # str_extract(selection[,2], '\\w*')), + text = selected_text, + textposition = "top right" , + #textfont_size = 4, + textfont = list(family = "sans serif", + size = 10, + color = "blue" + ) + ) + ) + + # p <- plotlyProxy(outputId = "res_volplot", session = session)$x$data + # saveRDS(object = p, file = "p_proxy_build.rds") + + global_data$trace_count <- global_data$trace_count + 1 + #print(global_data$trace_count) + #output$debug_console <- renderPrint(global_data$trace_count) + + #output$debug_console <- renderPrint(selection) + + #update the selection column of the selected rows + #was designed so that selected rows can then be sorted on to move them to + #the top. however, the code currently invokes a perpetual looping update + #of the table and doesn't allow the selection of more than one row either + #probably due to reactivity of the row selection event -- maybe try + #binding to an action button instead of automatic updates via row + #selection? ---- code is now fixed + changes <- data.frame(row = selected_rows, + #col = 6, #column index of "selected" (index starts at 0) + col = 7, + value = "Yes") + + DT::dataTableProxy("res_table") %>% + DT::editData(data = isolate(global_data$dt), + info = changes, + rownames = F, + resetPaging = F, + clearSelection = "none") + + } + #else if the last row has been deselected, then reset selection column + else if(!is.null(input$res_table_row_last_clicked)){ + + changes <- data.frame(row = isolate(input$res_table_row_last_clicked), + #col = 6, #column index of "selected" (index starts at 0) + col = 7, + value = "No") + + DT::dataTableProxy("res_table") %>% + DT::editData(data = isolate(global_data$dt), + info = changes, + rownames = F, + resetPaging = F, + clearSelection = "all") + } + + }) + + + #resetting rows selected in the results table + observeEvent(input$ResetTable, { + proxy <- dataTableProxy("res_table") + + proxy %>% selectRows(NULL) + + output$res_colplot <- NULL + + #removing traces from the plotly volcano plot + # plotly traces are zero indexed so min count is global_data$base_traces-1 + if(global_data$trace_count > global_data$base_traces-1){ + plotlyProxy(outputId = "res_volplot", session = session) %>% + plotlyProxyInvoke("deleteTraces", + #as.list(c(2:global_data$trace_count)) + as.list(c( + global_data$base_traces:global_data$trace_count)) + ) + + global_data$trace_count <- global_data$base_traces-1 + #print(global_data$trace_count) + } + # if(global_data$trace_count > 2){ + # plotlyProxy(outputId = "res_volplot", session = session) %>% + # plotlyProxyInvoke("deleteTraces", + # #as.list(c(2:global_data$trace_count)) + # as.list(c(3:global_data$trace_count)) + # # as.list(c( + # # (global_data$trace_count-1):global_data$trace_count)) + # ) + # + # global_data$trace_count <- 2 + # } + + }) + + + #download the complete results + output$DownloadTableAll <- downloadHandler(filename = "Results.tab", + content = function(file){ + write.table( + x = global_data$result, + file = file, + quote = F, + sep = "\t", + col.names = T, + row.names = F) + }) + + #download the filtered results + output$DownloadTableFilt <- downloadHandler( + filename = "Results_filtered.tab", + content = function(file){ + write.table(x = + global_data$result[input$res_table_rows_all, , drop = F], + file = file, + quote = F, + sep = "\t", + col.names = T, + row.names = F) + }) + + #download the selected rows only + output$DownloadTableSelected <- downloadHandler( + filename = "Results_selected.tab", + content = function(file){ + write.table(x = global_data$result[ + input$res_table_rows_selected, , drop = F], + file = file, + quote = F, + sep = "\t", + col.names = T, + row.names = F) + }) + + #(de)select rows in the results table by (de)selecting it in volcano plot + observe({ + + #the req() circumvents the whole exists() and !is.null requirements in the + #outer if-else block below, reducing the check to just the 50 hard limit + req(event_data("plotly_click", + source = "volplot", + priority = "event")) + + volplot_selected <- event_data("plotly_click", + source = "volplot", + priority = "event") + proxy <- dataTableProxy("res_table") + + already_selected <- isolate(input$res_table_rows_selected) + # proxy %>% selectRows(c(volplot_selected$key, already_selected)) + + #the following code is designed to allow deselection, but it is bugged for + #some reason and requires the user to click the last selected node twice + #to deselect it will need to work on this in later updates if required as + #a feature ----- the bug is resolved by 'priority = "event"' above + #Also, hard limiting to max selections to 50 + # if(exists("volplot_selected") & !is.null(volplot_selected) & + # length(already_selected) < 50){ + if(length(already_selected) < 50){ + + if(length(intersect(volplot_selected$key, already_selected)) > 0){ + #print("entering 'if' block") + if(length(already_selected) > 1){ + #print("entering 'if>if' block") + proxy %>% selectRows(as.numeric( + setdiff(already_selected, volplot_selected$key))) + } + else{ + #print("entering 'if>else' block") + proxy %>% selectRows(NULL) + } + } + else if(length(union(volplot_selected$key, already_selected)) > 0){ + #print("entering 'else if' block") + proxy %>% selectRows(as.numeric( + union(volplot_selected$key, already_selected))) + } + # else if(length(volplot_selected$key) > 0 & + # length(already_selected) < 1){ + # proxy %>% selectRows(as.numeric(volplot_selected$key)) + # } + # else{ + # print("entering 'else' block") + # proxy %>% selectRows(NULL) + # } + + } + + #volplot_selected <- NULL + #print(isolate(global_data$trace_count)) + + }) + + + #creating reactive expression to group multiple inputs for the observeEvent + plotFormatAndSize <- reactive({ + list(input$SavePlotFormat, input$WidthPixels, input$HeightPixels) + }) + #change the snapshot save format and size of plotly plots + observeEvent(plotFormatAndSize(), { + + #formatting the input + format <- tolower(input$SavePlotFormat) + width <- input$WidthPixels + height <- input$HeightPixels + + #updating the format for the volcano plot + plotlyProxy(outputId = "res_volplot", session = session) %>% + plotlyProxyInvoke("reconfig", + displaylogo = F, + displayModeBar = T, + toImageButtonOptions = + list(format = format, + width = width, + height = height + ), + modeBarButtonsToRemove = list( + "sendDataToCloud", "zoom2d", "zoomIn2d", + "zoomOut2d", "pan2d", "select2d", "lasso2d", + "autoScale2d", "hoverClosestCartesian", + "hoverCompareCartesian" + ) + ) + + #updating the format for the counts plot + plotlyProxy(outputId = "res_colplot", session = session) %>% + plotlyProxyInvoke("reconfig", + displaylogo = F, + displayModeBar = T, + toImageButtonOptions = + list(format = format, + width = width, + height = height + ), + modeBarButtonsToRemove = list( + "sendDataToCloud", "zoom2d", "zoomIn2d", + "zoomOut2d", "pan2d", "select2d", "lasso2d", + "autoScale2d", "toggleSpikelines"#, + #"hoverClosestCartesian", + #"hoverCompareCartesian" + ) + ) + + }) + + + #display PCA, t-SNE and UMAP plots + observeEvent(input$TriplePlot, { + + output$PCAplot1 <- NULL + output$PCAplot2 <- NULL + output$PCAplot3 <- NULL + + # output$debug_console <- renderPrint(input$TriplePlot == "PCA") + + if(input$TriplePlot == "PCA"){ + #output$debug_console <- renderText(class(input$TriplePlot)) + #output$PCAplot1 <- renderPlotly({ + output$PCAplot1 <- renderPlot({ + #ggplotly(PCA_plots[[1]]) %>% toWebGL() + PCA_plots[[1]] + }) + output$PCAplot2 <- renderPlot({ + PCA_plots[[2]] + }) + output$PCAplot3 <- renderPlot({ + PCA_plots[[3]] + }) + } + else if(input$TriplePlot == "t-SNE"){ + output$PCAplot1 <- renderPlot({ + tSNE_plots[[1]] + }) + output$PCAplot2 <- renderPlot({ + tSNE_plots[[2]] + }) + output$PCAplot3 <- renderPlot({ + tSNE_plots[[3]] + }) + } + else if(input$TriplePlot == "UMAP"){ + output$PCAplot1 <- renderPlot({ + UMAP_plots[[1]] + }) + output$PCAplot2 <- renderPlot({ + UMAP_plots[[2]] + }) + output$PCAplot3 <- renderPlot({ + UMAP_plots[[3]] + }) + } + else if(input$TriplePlot == "PCA - tissue means"){ + output$PCAplot1 <- renderPlot({ + PCA_means_plots[[1]] + }) + output$PCAplot2 <- renderPlot({ + PCA_means_plots[[2]] + }) + output$PCAplot3 <- renderPlot({ + PCA_means_plots[[3]] + }) + } + + }) + + + # display TCR chord diagrams ---- + observeEvent(input$TCRChordSelect, { + output$TCRChordImage <- renderImage(expr = { + + img <- readPNG( + source = paste0("www/TCR_plots/", + TCR_plots_list[grep(pattern = input$TCRChordSelect, + x = TCR_plots_list, + ignore.case = T)]), + native = T, info = T) + + list(src = paste0("www/TCR_plots/", + TCR_plots_list[grep(pattern = input$TCRChordSelect, + x = TCR_plots_list, + ignore.case = T)]), + contentType = 'image/png', + width = dim(img)[2]*0.15, + height = dim(img)[1]*0.15#, + #alt = "This is alternate text" + ) + }, deleteFile = F) + }) + + tsne_aging_img <- function(filename){ + img <- readPNG(source = filename, native = T, info = T) + list( + src = filename, + contentType = 'image/png', + width = dim(img)[2]*0.5, + height = dim(img)[1]*0.5 + ) + } + + observeEvent(input$browser, browser()) + + # aging tSNE plots ---- + observeEvent(input$agingTSNEtissue, { + tissue_files <- paste0("www/Aging_tsne_plots/", + aging_tsne_plots_list[grep(pattern = input$agingTSNEtissue, + x = aging_tsne_plots_list, + ignore.case = T)]) + tissue_files <- tissue_files[c(6,2,3,4,5,1)] + # ordering them manually + + output$agingTSNEImage1 <- renderImage( + expr = tsne_aging_img(tissue_files[2]), deleteFile = F + ) + + output$agingTSNEImage2 <- renderImage( + expr = tsne_aging_img(tissue_files[2]), deleteFile = F + ) + + }) + + + #display Pathview KEGG Pathways + observeEvent(input$PathviewPlotSelect, { + output$PathviewPlotImage <- renderImage(expr = { + + pathway_id <- gsub(pattern = " - .*", + replacement = "", + x = input$PathviewPlotSelect, + ignore.case = T) + + img <- readPNG( + source = paste0("www/Pathview_plots/", + Pathview_plots_list[grep(pattern = pathway_id, + x = Pathview_plots_list, + ignore.case = T)]), + native = T, info = T) + + list(src = paste0("www/Pathview_plots/", + Pathview_plots_list[grep(pattern = pathway_id, + x = Pathview_plots_list, + ignore.case = T)]), + contentType = 'image/png', + width = dim(img)[2],#*0.35, + height = dim(img)[1]#*0.35#, + #alt = "This is alternate text" + ) + }, deleteFile = F) + }) + + #display Pathview Legend and Legend2 + output$PathviewPlotLegend <- renderImage(expr = { + + img <- readPNG(source = "www/Legend.png", + native = T, info = T) + + list(src = "www/Legend.png", + contentType = 'image/png', + width = dim(img)[2], + height = dim(img)[1]#, + #alt = "This is alternate text" + ) + }, deleteFile = F) + output$PathviewPlotLegend2 <- renderImage(expr = { + + img <- readPNG(source = "www/Legend2.png", + native = T, info = T) + + list(src = "www/Legend2.png", + contentType = 'image/png', + width = dim(img)[2]*0.6, + height = dim(img)[1]*0.6#, + #alt = "This is alternate text" + ) + }, deleteFile = F) + + #download Pathview pathways + output$DownloadPathviewSelected <- downloadHandler( + filename = function() { + paste0(input$PathviewPlotSelect, ".zip") + }, + #filename = paste0(input$PathviewPlotSelect, ".zip"), + content = function(file){ + + pathway_id <- gsub(pattern = " - .*", + replacement = "", + x = input$PathviewPlotSelect, + ignore.case = T) + + zip(zipfile = file, + files = c(paste0("www/Pathview_plots/", + Pathview_plots_list[grep(pattern = pathway_id, + x = Pathview_plots_list, + ignore.case = T)]), + "www/Legend.png", "www/Legend2.png"), + flags = "-j") + }, + contentType = "application/zip") + # output$DownloadPathviewSelected <- downloadHandler( + # filename = paste0(input$PathviewPlotSelect, ".png"), + # content = function(file){ + # + # pathway_id <- gsub(pattern = " - .*", + # replacement = "", + # x = input$PathviewPlotSelect, + # ignore.case = T) + # + # file.copy(paste0("www/Pathview_plots/", + # Pathview_plots_list[grep(pattern = pathway_id, + # x = Pathview_plots_list, + # ignore.case = T)]), + # file, + # copy.mode = F, + # copy.date = F + # ) + # }, + # contentType = "image/png") + output$DownloadPathviewAll <- downloadHandler( + filename = function() { + paste0("all_pathways.zip") + }, + #filename = "all_pathways.zip", + content = function(file){ + + zip(zipfile = file, + files = c(paste0("www/Pathview_plots/", Pathview_plots_list), + "www/Legend.png", "www/Legend2.png"), + flags = "-j") + + }, + contentType = "application/zip") + + #download TCR plots + TCR_plots_list + output$DownloadTCRSelected <- downloadHandler( + filename = function() { + paste0(input$TCRChordSelect, "_chord_diagram.png") + }, + #filename = paste0(input$TCRChordSelect, "_chord_diagram.png"), + content = function(file){ + + file.copy(paste0("www/TCR_plots/", + TCR_plots_list[grep(pattern = input$TCRChordSelect, + x = TCR_plots_list, + ignore.case = T)]), + file, + copy.mode = F, + copy.date = F + ) + }, + contentType = "image/png") + output$DownloadTCRAll <- downloadHandler( + filename = function() { + paste0("all_chord_diagrams.zip") + }, + #filename = "all_chord_diagrams.zip", + content = function(file){ + + zip(zipfile = file, + files = paste0("www/TCR_plots/", TCR_plots_list), + flags = "-j") + + }, + contentType = "application/zip") + + #downloading Supplementary Materials + output$MultiQCBulk <- downloadHandler( + filename = function() { + paste0("QC and pre-processing report.zip") + }, + content = function(file){ + file.copy("www/MultiQC_report/QC and pre-processing report.zip", + file, + copy.mode = F, + copy.date = F + ) + }, + contentType = "application/zip") + output$GlobalBulk <- downloadHandler( + filename = function() { + paste0("PCA, t-SNE and UMAP plots.zip") + }, + content = function(file){ + zip(zipfile = file, + files = paste0("www/Global_bulk/", + list.files(path = "www/Global_bulk/")), + flags = "-j" + ) + }, + contentType = "application/zip") + #"GSEABulk", DiffExprsBulk + output$DiffExprsBulk <- downloadHandler( + filename = function() { + paste0("Counts and differential expression table.xlsx") + }, + content = function(file){ + file.copy("www/DESeq2_table/output_table.xlsx", + file, + copy.mode = F, + copy.date = F + ) + }, + contentType = "xlsx") + output$GSEABulk <- downloadHandler( + filename = function() { + paste0("GSEA table.xlsx") + }, + content = function(file){ + file.copy("www/GSEA_table/output_gage.xlsx", + file, + copy.mode = F, + copy.date = F + ) + }, + contentType = "xlsx") + + + #dynamic tutorials for rintrojs + { + steps <- reactive(data.frame(element = c("#Group1 + .selectize-control", + "#Group1Selector + .selectize-control", + "#Group1Add", + "#Group1Clear", + "#Group2 + .selectize-control", + "#AddOps", + "#TPCFilt + .selectize-control", + "#log2FCFilt", + "#AdjpFilt", + "#GenContrasts", + #"#res_table", + #"#DownloadTableAll", + #"#DownloadTableFilt", + #"#DownloadTableSelected", + #"#ResetTable", + "#mydropdown"), + intro = c(paste0("Start by selecting the ", + "tissue(s) that need to be ", + "contrasted. Selected ", + "tissue(s) can be removed ", + "by clicking on the name ", + "and pressing the 'backspace' ", + "or 'delete' key."), + paste0("Groups of tissues can be ", + "selected in bulk from ", + "here..."), + paste0("...and added using this ", + "button."), + paste0("The whole selection can ", + "be cleared by clicking ", + "this button."), + paste0("Tissue(s) to be contrasted ", + "against can be selected ", + "here. Note that tissues ", + "already selected in the ", + "first group are not ", + "available here and vice ", + "versa."), + paste0("This checkbox shows and ", + "hides additional options ", + "for the analysis."), + paste0("Transcripts per cell ", + "filtering filters out low ", + "expression genes from the ", + "results and plots. The ", + "filtering criteria is at ", + "least the selected amount ", + "of counts per cell in at ", + "least one Treg tissue."), + paste0("The minimum absolute log2 ", + "fold change for a gene to ", + "be considered ", + "differentially expressed."), + paste0("The maximum adjusted p.", + "value for a gene to be ", + "considered differentially ", + "expressed."), + paste0("Clicking this button will ", + "generate the contrasts ", + "between the groups ", + "selected above. This ", + "button remains disabled ", + "till both groups have at ", + "least one tissue selected."), + #paste0(), + paste0("This button can be ", + "clicked to show a summary ", + "of the instructions ", + "and features.")))) + + observeEvent(input$TutorialButton,{ + updateCheckboxInput(session = session, + inputId = "AddOps", + value = TRUE) + + introjs(session = session , options = list(steps=steps())) + }) + + steps2 <- reactive(data.frame(element = c("#res_table", + "#DownloadTableAll", + "#DownloadTableFilt", + "#DownloadTableSelected", + "#ResetTable", + "#PlotSaveOps", + "#SavePlotFormat", + "#WidthPixels", + "#HeightPixels", + "#res_volplot" + ), + intro = c(paste0("The results of the ", + "contrast are tabulated ", + "here. Column headers can ", + "be clicked to sort by ", + "that column (sorting can ", + "be done on multiple ", + "columns by pressing the ", + "'shift' key and clicking ", + "on the other header). ", + "Columns can be searched ", + "or filtered by clicking ", + "on the blank fields ", + "under the column headers.", + " Clicking a row will ", + "select the row in the ", + "table and highlight the ", + "respective gene in the ", + "volcano plot (upto 50 ", + "rows can be selected by ", + "pressing the 'shift' or ", + "'control' key)."), + paste0("Download the complete ", + "contrast results as a ", + "table."), + paste0("Download only the rows ", + "which are left after ", + "filtering (empty table if", + " no filtering done)."), + paste0("Download only the rows ", + "which have been manually ", + "selected (empty table if", + " no selection done)."), + paste0("This button resets any ", + "rows that have been ", + "selected."), + paste0("This checkbox shows and ", + "hides options related ", + "to exporting the plots."), + paste0("Select the format for ", + "exporting the plot."), + paste0("Specify the width of the ", + "plot in pixels for export."), + paste0("Specify the height of the ", + "plot in pixels for export."), + paste0("The interactive volcano ", + "plot of the contrast. ", + "Hovering over a data ", + "point would show a ", + "tooltip with additional ", + "information. Clicking on ", + "a data point will select ", + "the respective gene in ", + "the table and will also ", + "generate a column plot to ", + "visualise the expression ", + "of the selected gene in ", + "tissues. The plot can be ", + "zoomed-in by clicking and ", + "dragging a box over the ", + "area to be zoomed. ", + "Additionally the axes of ", + "the plot can be adjusted ", + "by dragging the edges of ", + "the respective axes. The ", + "zoom and/or axes can be ", + "reset by double-clicking ", + "in the empty space in the ", + "plot area or by clicking ", + "the house icon in the ", + "top-right of the plot. ", + "The plot can be exported ", + "using the export options ", + "by clicking the camera ", + "icon in the top-right of ", + "the plot. The same ", + "functionality except for ", + "data point selection also ", + "exists for the column ", + "plot.") + ))) + + observeEvent(input$TutorialButton2,{ + updateCheckboxInput(session = session, + inputId = "PlotSaveOps", + value = TRUE) + introjs(session = session , options = list(steps=steps2())) + }) + } + + } # closing bracket for server call +} + +# Create Shiny app ---- +shinyApp(ui = ui, server = server) + + + + + + From 4e61e95815e4272736cb52dbfe930fc78b6b6fb1 Mon Sep 17 00:00:00 2001 From: Laura Biggins Date: Tue, 29 Aug 2023 10:50:06 +0100 Subject: [PATCH 02/15] Adding the aging t-sne plots --- .../tsne_plot_Adrenals 100-week__cluster.png | Bin 0 -> 19499 bytes .../tsne_plot_Adrenals 12-week__cluster.png | Bin 0 -> 14151 bytes .../tsne_plot_Adrenals 20-week__cluster.png | Bin 0 -> 14288 bytes .../tsne_plot_Adrenals 30-week__cluster.png | Bin 0 -> 12818 bytes .../tsne_plot_Adrenals 52-week__cluster.png | Bin 0 -> 15127 bytes .../tsne_plot_Adrenals 8-week__cluster.png | 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zOpC&s4A;9&r%(3eDf<%|3sFbL&H2@jbW?>+GzqjrlNoMzBElS=M_!;+4>$gU4F`v! z1t>zh+|`?A+uHK%H1@Fr{!c48l0{yY{H=>G6S4d1oOpMLT%+kapl2cxAXj6!$euGC zv)5F4MbutNmIAL`F=&8ODK?|DOa)rYEC1$c2BI@m$ zvmf!zx0k`DJ##RkJ~4=nhiM-t-|^8f}IlRBO0^@otF&^ z9G%xTwHi_{5eiw<<(aj$u0ZF5Nf|JoC3-mDOCa_2ZBnL(T1r1dPtV~3X-;|vpzxHg z$By@#jHh(jp5VLC;%iE0KL5Pf>2;g|y~N#POirr@5Cd7nCT~Cmv&q zU7MLDRDjK(@_SIDp_Y=-T92&*#M-|Ein?zEFr^B2#3UuIb}%E3Q+NW5i&e0}g$|#T zT56q>@LC6>g1Xzf_%NSd$D5}H&-h<6=^5u@78f%{lIH}X_} zrzXn^Ook47+<*&};djX)cOgozomErgn8y?Re1mZ8#y5ne?(WiY;j&c0@q$XvZSU@m|2L4T<7E9kzZmPEPNX70S;tzlwaOt0GA)~5 z7jFmOYfX%Tu Date: Tue, 29 Aug 2023 10:51:42 +0100 Subject: [PATCH 03/15] Updated app.R to include the latest version from the shiny server that had updated descriptions. --- expressionViewer/app.R | 951 +++++++++++++++++++++++------------------ 1 file changed, 529 insertions(+), 422 deletions(-) diff --git a/expressionViewer/app.R b/expressionViewer/app.R index f6d5e0c..a04e91f 100644 --- a/expressionViewer/app.R +++ b/expressionViewer/app.R @@ -1,5 +1,7 @@ +# Developed by Samar H.K. Tareen + #importing libraries & functions, reading in data, and setting up global options { @@ -16,6 +18,7 @@ library(DT) library(ggplot2) library(ggrepel) + library(patchwork) library(viridis) library(plotly) library(stringr) @@ -32,9 +35,6 @@ } - # #functions and scripts - # source("customFunctions.R") - #data { #deseq2 object @@ -187,7 +187,7 @@ filtered_genes_lists <- readRDS(file = "data/Filtered_Gene_Lists.rds") #importing TCR plots - TCR_plots_list <- list.files(path = "www/TCR_plots/", pattern = ".png") + TCR_plots_list <- list.files(path = "www/TCR_plots/", pattern = "\\.png$") #print(TCR_plots_list) ## aging_tsne_plots_list ---- @@ -197,8 +197,8 @@ #importing Pathview plots { - Pathview_plots_list <- list.files(path = "www/Pathview_plots", - pattern = ".multi\\.png") + Pathview_plots_list <- list.files(path = "www/Pathview_plots/", + pattern = "\\.multi\\.png$") #print(Pathview_plots_list) #mapping Pathview plot files to pathway names @@ -231,7 +231,7 @@ # using shinyjs to enable or disable certain sections/elements of the app shinyjs::useShinyjs(), # using shinyalert to enable popup messages - shinyalert::useShinyalert(), + # shinyalert::useShinyalert(), # Step-by-step tutorial introjsUI(), @@ -250,8 +250,7 @@ #dashboardBody(), # App title ---- - #titlePanel("Expression Viewer"), - titlePanel("The Tissue Tregs Project"), + titlePanel("The Tissue Treg Project"), tabsetPanel( @@ -259,12 +258,25 @@ { tabPanel(title = "The Study", value = "TheStudy", - h3("General description/information of the study/data"), + h3("Transcriptional analysis of tissue-resident regulatory T cells"), - p(paste0("Some description of the study, as well as where to ", - "find additional sources such as papers and data.")), - p(paste0("Ask Adrian and Oliver as they would be able to ", - "provide a much better description")), + "A transcriptional analysis of tissue-resident ", + "regulatory T cells (Tregs) as part of the Tissue ", + "Treg Project from the ", + a("Liston-Dooley lab.", href='https://www.listonlab.uk/', + target = "_blank"), + "The project is based on Tregs and conventional ", + "T cells (Tconv) isolated from blood, spleen, ", + "lymph nodes, kidney, liver, lung, pancreas, and ", + "gut (Peyer’s patches, LPLs, IELs). The ", + "expression viewer, allowing interactive custom ", + "analysis and data download, was developed by ", + "Samar Tareen, is ", + a("open source,", href = 'https://github.com/AdrianListon/ExpressionViewer', + target = "_blank"), + "and is hosted by the Babraham Institute ", + a("Bioinformatics Group.", href = 'https://www.bioinformatics.babraham.ac.uk/', + target = "_blank"), hr(), @@ -316,11 +328,8 @@ br(), fluidRow( - h6(paste0("Copyright 2020, Liston Lab, Babraham Institute. ", - "Babraham internal/intranet release only. ", - "Copyright licence will most likely change by the ", - "first public release as per the requirements of ", - "the publication journal and BI.")) + h6(paste0("Copyright 2023, Liston-Dooley Lab, ", + "Babraham Institute and University of Cambridge.")) ) ) }, @@ -329,13 +338,36 @@ { tabPanel(title = "Global Analysis", value = "GlobalAnalysis", - h3("Exploratory analysis of the expression data"), + h3("Global analysis of tissue Treg transcriptome across different tissue sources"), - p(paste0("PCA, t-SNE and UMAP analysis and plots of the ", - "RNA-seq expression data from the study.")), + p(paste0("For the bulk RNA-seq analysis, Tregs and Tconv were ", + "isolated from blood, and perfused spleen, lymph ", + "nodes, kidney, liver, lung, pancreas, and gut ", + "(Peyer’s patches, LPLs, IELs) samples. Samples were ", + "prepared from Foxp3Thy1.1 reporter mice, allowing ", + "purification based on Foxp3 expression. mRNA was ", + "sequencing using the QuantSeq 3’mRNA-Seq Library ", + "Prep Kit for Illumina and the QuantSeg data analysis ", + "workflow. The relationship between samples at the ", + "global level is visualised through PCA (using all ", + "samples or tissue means), tSNE and UMAP projections.")), - p(paste0("Plots can be downloaded by right clicking the ", - "plot and clicking on 'Save image as...'")), + # p(paste0("For the single cell RNA-seq analysis, Tregs were ", + # "flow sorted from Foxp3Thy1.1 reporter mice, ", + # "pre-injected with anti-CD45 via intravenous ", + # "delivery. Cells were purified from blood, kidney, ", + # "liver, pancreas and LPLs from the gut. Purified ", + # "cells were CD4+Foxp3Thy1.1+ as well as negative for ", + # "intravenous CD45 labelling and for the exclusion ", + # "markers CD19, CD11b, CD8 and F4/80. Cells were ", + # "labelled with Hastag TotalSeq and loaded onto the ", + # "10x Chromium Controller with sequencing performed ", + # "on Illumina HiSeq. Single cell data is visually ", + # "presented as tSNE and UMAP projections, with each ", + # "tissue origin labelled a different colour.")), + + p(paste0("Plots may be downloaded by right clicking the ", + "plot and selecting 'Save image as...'")), #br(), hr(), @@ -382,11 +414,11 @@ hr(), - h4("Single cell RNA-seq data"), - - p("Will be available soon") + # h4("Single cell RNA-seq data"), + # + # p("Will be available soon") - ) + ) }, #Bulk RNA-seq Expression panel @@ -395,26 +427,43 @@ value = "BulkDiffExprs", fluidRow( - - column(width = 7, - h3("Generate contrasts"), - p(paste0("The differential expression analysis is performed ", - "using DESeq2. For a quick walkthrough on doing the ", - "the analysis, please click the yellow button with ", - "the question mark.")) - ), - - column(width = 5, - br(), - br(), - # actionButton(inputId = "TutorialButton", - # label = "Start tutorial"), - actionBttn(inputId = "TutorialButton", - label = "Start tutorial", - style = "material-circle", - color = "warning", - icon = icon("question")), - ) + + column(width = 7, + h3("Generate contrasts"), + + "An interactive differential analysis tool based ", + "on the bulk RNAseq dataset. Pick any population ", + "or group of populations to run a differential ", + "expression analysis by comparison to any other ", + "population or group of populations. Both Tregs ", + "and Tconv are available, as purified from the ", + "blood, spleen, lymph nodes, kidney, liver, lung, ", + "pancreas, Peyer’s Patches, and intraepithelial ", + "cells (IEL) and lamina propria lymphocytes (LPL) ", + "from the intestines. The differential expression ", + "analysis is performed using DESeq2, using a custom ", + "script available on ", + a("GitHub.", href= 'https://github.com/AdrianListon/TissueTregs', + target = "_blank"), + p(), + "For a quick walkthrough on doing the analysis, ", + "please click the yellow button with the question ", + "mark. Download options for visualization and ", + "data tables become available after running a contrast.", + + ), + + column(width = 5, + br(), + br(), + # actionButton(inputId = "TutorialButton", + # label = "Start tutorial"), + actionBttn(inputId = "TutorialButton", + label = "Start tutorial", + style = "material-circle", + color = "warning", + icon = icon("question")), + ) ), hr(), @@ -474,7 +523,7 @@ "duplicate ", "tissues that are ", "already selected." - )), + )), h5(paste0("Clicking the ", "'Generate ", @@ -489,26 +538,26 @@ "results and a ", "volcano plot on ", "right." - )), + )), h5(paste0("Clicking the ", - "'Show Advanced Options' ", - "checkbox reveals three ", - "additional options to ", - "adjust the analysis and ", - "visualisation. The ", - "'Minimum Transcripts per ", - "Cell' drop down list ", - "gives options to filter ", - "results to remove low ", - "expression genes if ", - "needed. The 'log2 Fold ", - "Change' and 'Adjusted ", - "p.value' fields are the ", - "significance criteria ", - "used in the volcano plot ", - "and can be adjusted as ", - "needed." + "'Show Advanced Options' ", + "checkbox reveals three ", + "additional options to ", + "adjust the analysis and ", + "visualisation. The ", + "'Minimum Transcripts per ", + "Cell' drop down list ", + "gives options to filter ", + "results to remove low ", + "expression genes if ", + "needed. The 'log2 Fold ", + "Change' and 'Adjusted ", + "p.value' fields are the ", + "significance criteria ", + "used in the volcano plot ", + "and can be adjusted as ", + "needed." )), hr(), @@ -635,7 +684,7 @@ "specifying the ", "height and width ", "in pixels." - )) + )) ) ), ), @@ -652,7 +701,7 @@ ignore.case = T)], Tconv = results_names[ grepl(pattern = "cellTypeNT", - #grepl(pattern = "_Tconv", + #grepl(pattern = "_Tconv", x = results_names, ignore.case = T)] ), @@ -664,40 +713,40 @@ "to manually", " select ", "...") - ) + ) #) ), #fluidRow( - #h5("Select available:"), - #h5("Quick add available:"), - # actionButton(inputId = "Group1SelectTconv", - # label = "Tconv"), - # actionButton(inputId = "Group1SelectTregs", - # label = "Tregs"), - selectizeInput(inputId = "Group1Selector", - label = "Add available:", - choice = list( - Treg = c("All Tregs", - "Blood Tregs", - "Lymphoid Tregs", - "Non-lymphoid Tregs", - "Gut-associated Tregs"), - Tconv = c("All Tconv", - "Blood Tconv", - "Lymphoid Tconv", - "Non-lymphoid Tconv", - "Gut-associated Tconv") - ), - selected = "All Tregs", - multiple = F, - options = list( - placeholder = "Click to select") - ), - actionButton(inputId = "Group1Add", - label = "Add"), - actionButton(inputId = "Group1Clear", - label = "Clear Selection"), + #h5("Select available:"), + #h5("Quick add available:"), + # actionButton(inputId = "Group1SelectTconv", + # label = "Tconv"), + # actionButton(inputId = "Group1SelectTregs", + # label = "Tregs"), + selectizeInput(inputId = "Group1Selector", + label = "Add available:", + choice = list( + Treg = c("All Tregs", + "Blood Tregs", + "Lymphoid Tregs", + "Non-lymphoid Tregs", + "Gut-associated Tregs"), + Tconv = c("All Tconv", + "Blood Tconv", + "Lymphoid Tconv", + "Non-lymphoid Tconv", + "Gut-associated Tconv") + ), + selected = "All Tregs", + multiple = F, + options = list( + placeholder = "Click to select") + ), + actionButton(inputId = "Group1Add", + label = "Add"), + actionButton(inputId = "Group1Clear", + label = "Clear Selection"), #), #br(), @@ -734,39 +783,39 @@ "to manually", " select ", "...") - ) + ) ), #fluidRow( - #h5("Select available:"), - #h5("Quick add available:"), - # actionButton(inputId = "Group2SelectTconv", - # label = "Tconv"), - # actionButton(inputId = "Group2SelectTregs", - # label = "Tregs"), - selectizeInput(inputId = "Group2Selector", - label = "Add available:", - choice = list( - Treg = c("All Tregs", - "Blood Tregs", - "Lymphoid Tregs", - "Non-lymphoid Tregs", - "Gut-associated Tregs"), - Tconv = c("All Tconv", - "Blood Tconv", - "Lymphoid Tconv", - "Non-lymphoid Tconv", - "Gut-associated Tconv") - ), - selected = "All Tconv", - multiple = F, - options = list( - placeholder = "Click to select") - ), - actionButton(inputId = "Group2Add", - label = "Add"), - actionButton(inputId = "Group2Clear", - label = "Clear Selection"), + #h5("Select available:"), + #h5("Quick add available:"), + # actionButton(inputId = "Group2SelectTconv", + # label = "Tconv"), + # actionButton(inputId = "Group2SelectTregs", + # label = "Tregs"), + selectizeInput(inputId = "Group2Selector", + label = "Add available:", + choice = list( + Treg = c("All Tregs", + "Blood Tregs", + "Lymphoid Tregs", + "Non-lymphoid Tregs", + "Gut-associated Tregs"), + Tconv = c("All Tconv", + "Blood Tconv", + "Lymphoid Tconv", + "Non-lymphoid Tconv", + "Gut-associated Tconv") + ), + selected = "All Tconv", + multiple = F, + options = list( + placeholder = "Click to select") + ), + actionButton(inputId = "Group2Add", + label = "Add"), + actionButton(inputId = "Group2Clear", + label = "Clear Selection"), #), hr(), @@ -828,19 +877,19 @@ br(), fluidRow( column(width = 10, - p(paste0("For an overview of the results, ", - "features and interactability, please ", - "click the yellow button to the right.")) + p(paste0("For an overview of the results, ", + "features and interactability, please ", + "click the yellow button to the right.")) ), - column(width = 2, - # actionButton(inputId = "TutorialButton2", - # label = "Start tutorial"), - actionBttn(inputId = "TutorialButton2", - label = "Start tutorial", - style = "material-circle", - color = "warning", - icon = icon("question")), - ) + column(width = 2, + # actionButton(inputId = "TutorialButton2", + # label = "Start tutorial"), + actionBttn(inputId = "TutorialButton2", + label = "Start tutorial", + style = "material-circle", + color = "warning", + icon = icon("question")), + ) ), hr(), @@ -850,64 +899,64 @@ conditionalPanel( condition = "input.PlotSaveOps", - - fluidRow( - column(width = 3,#) - radioGroupButtons(inputId = "SavePlotFormat", - label = HTML(paste0("Plot", - " Export ", - "Format")), - choices = c("SVG", "PNG", "JPEG"), - #SVG, PNG, JPEG and WEBP - #np native PDF support - selected = "SVG", - direction = "vertical", #"horizontal", - checkIcon = list( - # yes = icon("ok", - # lib = "glyphicon") - yes = icon(name = "file-image", - lib = "font-awesome") - ), - size = "normal" # xs/sm/normal/lg - ), - ), - column(width = 7,#) - includeScript(path = "js/validate-number.js"), - numericInputIcon(inputId = "WidthPixels", - label = HTML(paste0("Plot ", - "Size in ", - " Pixels ", #"
", - "(Range: 100 ", - "- 10,000) ", - #"

", - "
", - "Width")), #in Pixels", - value = 880, - min = 100, - max = 10000, - step = 10, - width = "85%", - icon = list( - icon(name = "ruler-horizontal", - lib = "font-awesome"), - "px") - ), - - numericInputIcon(inputId = "HeightPixels", - label = "Height", #in Pixels", - value = 562, - min = 100, - max = 10000, - step = 10, - width = "85%", - icon = list( - icon(name = "ruler-vertical", - lib = "font-awesome"), - "px") - ) + fluidRow( + column(width = 3,#) + radioGroupButtons(inputId = "SavePlotFormat", + label = HTML(paste0("Plot", + " Export ", + "Format")), + choices = c("SVG", "PNG", "JPEG"), + #SVG, PNG, JPEG and WEBP + #np native PDF support + selected = "SVG", + direction = "vertical", #"horizontal", + checkIcon = list( + # yes = icon("ok", + # lib = "glyphicon") + yes = icon(name = "file-image", + lib = "font-awesome") + ), + size = "normal" # xs/sm/normal/lg + ), + ), + column(width = 7,#) + includeScript(path = "js/validate-number.js"), + + numericInputIcon(inputId = "WidthPixels", + label = HTML(paste0("Plot ", + "Size in ", + " Pixels ", #"
", + "(Range: 100 ", + "- 10,000) ", + #"

", + "
", + "Width")), #in Pixels", + value = 880, + min = 100, + max = 10000, + step = 10, + width = "85%", + icon = list( + icon(name = "ruler-horizontal", + lib = "font-awesome"), + "px") + ), + + numericInputIcon(inputId = "HeightPixels", + label = "Height", #in Pixels", + value = 562, + min = 100, + max = 10000, + step = 10, + width = "85%", + icon = list( + icon(name = "ruler-vertical", + lib = "font-awesome"), + "px") + ) + ) ) - ) ), hr(), @@ -1002,8 +1051,23 @@ h3("Visualise enriched KEGG pathways"), - p(paste0("The results of the pathway analsis using GAGE and ", - "Pathview can be visualised below.")), + p(paste0("Tregs and Tconv were isolated from blood, and ", + "perfused spleen, lymph nodes, kidney, liver, lung, ", + "pancreas, and gut (Peyer’s patches, LPLs, IELs) ", + "samples. Samples were prepared from Foxp3Thy1.1 ", + "reporter mice, allowing purification based on Foxp3 ", + "expression. Bulk mRNA was sequencing using the ", + "QuantSeq 3’mRNA-Seq Library Prep Kit for Illumina ", + "and the QuantSeg data analysis workflow. Differential ", + "analysis was performed between Tregs in the blood, ", + "lymphoid tissues (spleen, lymph nodes), non-lymphoid ", + "organs (kidney, liver, lung, pancreas) and ", + "gut-associated tissues (Peyer’s patches, LPLs, IELs). ", + "KEGG pathways enriched for differential expression ", + "are visualised using GAGE and Pathview, with each ", + "pairwise differential expression set visualised as ", + "log2 fold-change for any gene within the pathway with ", + "differential expression.")), hr(), @@ -1087,14 +1151,30 @@ # ) # }, - #TCR panel ---- + # TCR panel ---- { - tabPanel(title = "TCR", value = "TCR", + tabPanel(title = "TCR Repertoire", value = "TCR", h3("TCR repertoire analysis"), - p(paste0("Description of the TCR repertoire analysis - ", - "ask Orian")), + p(paste0("Tregs were flow sorted from Foxp3Thy1.1 reporter ", + "mice, pre-injected with anti-CD45 via intravenous ", + "delivery. Cells were purified from blood, kidney, ", + "liver, pancreas and LPLs from the gut. Purified ", + "cells were CD4+Foxp3Thy1.1+ as well as negative for ", + "intravenous CD45 labelling and for the exclusion ", + "markers CD19, CD11b, CD8 and F4/80. Cells were ", + "labelled with Hastag TotalSeq and loaded onto the ", + "10x Chromium Controller with sequencing performed on ", + "Illumina HiSeq. Data is visualised for each cell ", + "where TCRbeta CDR3 expression was observed using ", + "Chord diagrams. Chord diagrams represent amino ", + "acid-level unique clonotypes on the circular axis, ", + "with shared clonotypes represented on the radial ", + "axis as shared across different tissues in the same ", + "mouse (green), shared across different tissues in ", + "different mice (black) and shared between the same ", + "tissue in different mice (pink).")), #br(), hr(), @@ -1116,7 +1196,7 @@ multiple = F, options = list( placeholder = "Click to select") - ), + ), downloadButton( outputId = "DownloadTCRSelected", @@ -1128,7 +1208,7 @@ label = "All Tissues" ) - ), + ), column(width = 8, @@ -1138,11 +1218,12 @@ withSpinner(type = 3, size = 0.5, color.background = "#FFFFFF") - ) + ) ) - ) + ) }, + # Aging t-SNE panel ---- # It might be worth putting the pngs together into one plot { @@ -1154,7 +1235,7 @@ hr(), fluidRow( column( - width = 3, + width = 2, selectizeInput( inputId = "agingTSNEtissue", label = "Select Tissue", @@ -1162,7 +1243,7 @@ #selectize = T, multiple = FALSE, options = list( - placeholder = "Click to select") + placeholder = "Click to select") ), downloadButton( outputId = "DownloadAgingTSNEtissue", @@ -1174,30 +1255,37 @@ ), actionButton("browser", "browser") ), - column( - width = 4, - imageOutput( - outputId = "agingTSNEImage1", - width = 4344*0.15, - height = 4601*0.15 - ) %>% - withSpinner(type = 3, size = 0.5, color.background = "#FFFFFF") - ), - column( - width = 4, - imageOutput( - outputId = "agingTSNEImage2", - width = 4344*0.15, - height = 4601*0.15 - ) %>% - withSpinner(type = 3, size = 0.5, color.background = "#FFFFFF") - ) + flowLayout( + wellPanel( + verticalLayout( + textOutput(outputId = "agingTSNETitle1"), + imageOutput( + outputId = "agingTSNEImage1"#, + #width = 4344*0.15, + #height = 4601*0.15 + ) #%>% + #withSpinner(type = 3, size = 0.5, color.background = "#FFFFFF") + ) + ), + wellPanel( + verticalLayout( + textOutput(outputId = "agingTSNETitle2"), + imageOutput( + outputId = "agingTSNEImage2", + width = 4344*0.15, + height = 4601*0.15 + ) #%>% + #withSpinner(type = 3, size = 0.5, color.background = "#FFFFFF") + ) + ) + ) ) ) } ), ) ## ui fluid page ends at bracket immediately above + } @@ -1234,7 +1322,7 @@ left_log = 0, right_log = 0, first_run_done = F - ) + ) #live updating UI elements observe({ @@ -1250,7 +1338,7 @@ grepl(pattern = "cellTypeT", x = results_names, ignore.case = T) & !(results_names %in% - input$Group2)], + input$Group2)], Tconv = results_names[ grepl(pattern = "cellTypeNT", x = results_names, @@ -1258,7 +1346,7 @@ input$Group2)] ), selected = input$Group1) - + updateSelectInput(session = session, inputId = "Group2", label = "Contrast Group 2", @@ -1291,12 +1379,12 @@ 10000, ifelse( input$HeightPixels < 100, 100, round(input$HeightPixels)) - ) - ), + ) + ), min = 100, max = 10000, step = 10 - ) + ) updateNumericInput(session = session, inputId = "WidthPixels", @@ -1310,13 +1398,13 @@ 10000, ifelse( input$WidthPixels < 100, 100, round(input$WidthPixels)) - ) - ), + ) + ), min = 100, max = 10000, step = 10 - ) - + ) + updateNumericInput(session = session, inputId = "log2FCFilt", value = ifelse(input$log2FCFilt > 20, 20, @@ -1329,10 +1417,10 @@ input$log2FCFilt == "" | input$log2FCFilt == 0, 2, input$log2FCFilt) - ) - ), + ) + ), min = 0, max = 20, step = 0.1 - ) + ) updateNumericInput(session = session, inputId = "AdjpFilt", value = ifelse(input$AdjpFilt > 1, 1, @@ -1345,10 +1433,10 @@ input$AdjpFilt == "" | input$AdjpFilt == 0, 0.01, input$AdjpFilt) - ) - ), + ) + ), min = 0, max = 1, step = 0.01 - ) + ) #print(input$AdjpFilt) }) @@ -1395,7 +1483,7 @@ x = available, ignore.case = T) & !(available %in% input$Group2)]) - ) + ) } else if(group1_selection[1] == "Lymphoid"){ updateSelectInput(session = session, @@ -1418,7 +1506,7 @@ x = available, ignore.case = T) & !(available %in% input$Group2)]) - ) + ) } else if(group1_selection[1] == "Non-lymphoid"){ updateSelectInput(session = session, @@ -1441,7 +1529,7 @@ x = available, ignore.case = T) & !(available %in% input$Group2)]) - ) + ) } else if(group1_selection[1] == "Gut-associated"){ updateSelectInput(session = session, @@ -1464,7 +1552,7 @@ x = available, ignore.case = T) & !(available %in% input$Group2)]) - ) + ) } else{ updateSelectInput(session = session, @@ -1484,7 +1572,7 @@ ), selected = c(input$Group1, available[ !(available %in% input$Group2)]) - ) + ) } }) @@ -1644,7 +1732,7 @@ x = available, ignore.case = T) & !(available %in% input$Group1)]) - ) + ) } else if(group2_selection[1] == "Lymphoid"){ updateSelectInput(session = session, @@ -1860,7 +1948,7 @@ length(input$Group2) < 1) && (is.numeric(input$log2FCFilt) & is.numeric(input$AdjpFilt)) - ) + ) }) #output$show_panels <- reactive({FALSE}) #outputOptions(x = output, name = "show_panels", suspendWhenHidden = FALSE) @@ -1873,24 +1961,26 @@ #splash alert for the user informing them that the analysis may take some # time - if(!global_data$first_run_done){ - # sendSweetAlert(session = session, - # title = "Computing contrasts", - # text = paste0("The computation may take some time during ", - # "which the tool may appear to not be doing ", - # "anything. Once the results are computed ", - # "they will populate the tab."), - # type = "info") - shinyalert(title = "Computing contrasts", - text = paste0("The computation may take some time during ", - "which the tool may appear to not be doing ", - "anything. Once the results are computed, ", - "they will populate the tab."), - type = "info", - animation = F, #"slide-from-bottom", - inputId = "AlertFirstRunDone") - global_data$first_run_done <- T - } + # if(!global_data$first_run_done){ + # sendSweetAlert(session = session, + # title = "Computing contrasts", + # text = paste0("The computation may take some time during ", + # "which the tool may appear to not be doing ", + # "anything. Once the results are computed ", + # "they will populate the tab."), + # type = "info") + shinyalert(title = "Computing contrasts,\nplease wait...", + text = paste0("The contrasts may take some time to compute.\n", + "The results will populate the tab once ready."), + type = "info", + animation = F, #"slide-from-bottom", + inputId = "AlertFirstRunDone", + showCancelButton = FALSE, + showConfirmButton = FALSE, + closeOnEsc = FALSE, + closeOnClickOutside = FALSE) + # global_data$first_run_done <- T + # } #ensuring that the minimum log2FC and minimum adj.p.value fields weren't @@ -1927,7 +2017,7 @@ meta_data = MGI_info, mean_norm_counts = mean_norm_counts, tpc_filtered_list = tpc_filtered_list - ) + ) #print(class(result[[2]])) #global_data$result <- result @@ -2022,21 +2112,21 @@ #buttons = c('csv', 'excel', 'pdf'), #order = list(list(5, 'asc')) order = list(list(6, 'asc')) - ), + ), filter = list(position = "top", plain = F - ), + ), editable = F, rownames = F) #%>% - # DT::formatRound(columns = c("Mean.Counts"), - # digits = 2, - # mark = "") %>% - # DT::formatRound(columns = c("log2.Fold.Change"), - # digits = 3, - # mark = "") %>% - # DT::formatSignif(columns = c("Adjusted.p.value"), - # digits = 3, - # mark = "") + # DT::formatRound(columns = c("Mean.Counts"), + # digits = 2, + # mark = "") %>% + # DT::formatRound(columns = c("log2.Fold.Change"), + # digits = 3, + # mark = "") %>% + # DT::formatSignif(columns = c("Adjusted.p.value"), + # digits = 3, + # mark = "") }) #used for updating DataTable values for the 'Selected' column @@ -2077,7 +2167,7 @@ height = isolate(input$HeightPixels), log2FCFilt = input$log2FCFilt, AdjpFilt = input$AdjpFilt - ) + ) output$res_volplot <- renderPlotly({p}) p <- plotly_build(p) global_data$base_traces <- length(p$x$data) @@ -2106,6 +2196,19 @@ #resuming the DT row selection observer #res_table_row_observer$resume() + #remove popup shinyalert from earlier using second alert with immediate + # set to TRUE + shinyalert(title = "Computation Complete", + text = paste0("Results are being populated"), + type = "info", + animation = F, #"slide-from-bottom", + inputId = "RemoveAlert", + immediate = TRUE, timer = 1, + showCancelButton = FALSE, + showConfirmButton = FALSE, + closeOnEsc = FALSE, + closeOnClickOutside = FALSE) + }) @@ -2125,7 +2228,7 @@ #as.list(c(2:global_data$trace_count)) as.list(c( global_data$base_traces:global_data$trace_count)) - ) + ) global_data$trace_count <- global_data$base_traces-1 #print(global_data$trace_count) @@ -2152,7 +2255,7 @@ format = tolower(isolate(input$SavePlotFormat)), width = isolate(input$WidthPixels), height = isolate(input$HeightPixels) - ) + ) }) #adding traces to the plotly volcano plot @@ -2196,7 +2299,7 @@ replacement = "", x = selection[,1], ignore.case = T) - ) + ) #print(selected_text) } else{ @@ -2210,7 +2313,7 @@ replacement = "", x = selection[,2], ignore.case = T) - ) + ) #print(selected_text) } } @@ -2261,27 +2364,27 @@ plotlyProxy(outputId = "res_volplot", session = session) %>% plotlyProxyInvoke("addTrace", list(#x = c(selection[1,5],selection[,5]), - #y = -log10(c(selection[1,9], selection[,9])), - x = selected_x, - y = -log10(selected_y), - type = "scattergl", - name = "Selected genes", - mode = "markers+text", - marker = list(#symbol = "x", - symbol = "circle-open", - size = 10, - #colorscale = "Viridis" - color = "blue" - ), - # text = c(str_extract(selection[,2], '\\w*')[1], - # str_extract(selection[,2], '\\w*')), - text = selected_text, - textposition = "top right" , - #textfont_size = 4, - textfont = list(family = "sans serif", - size = 10, - color = "blue" - ) + #y = -log10(c(selection[1,9], selection[,9])), + x = selected_x, + y = -log10(selected_y), + type = "scattergl", + name = "Selected genes", + mode = "markers+text", + marker = list(#symbol = "x", + symbol = "circle-open", + size = 10, + #colorscale = "Viridis" + color = "blue" + ), + # text = c(str_extract(selection[,2], '\\w*')[1], + # str_extract(selection[,2], '\\w*')), + text = selected_text, + textposition = "top right" , + #textfont_size = 4, + textfont = list(family = "sans serif", + size = 10, + color = "blue" + ) ) ) @@ -2312,7 +2415,7 @@ rownames = F, resetPaging = F, clearSelection = "none") - + } #else if the last row has been deselected, then reset selection column else if(!is.null(input$res_table_row_last_clicked)){ @@ -2392,7 +2495,7 @@ sep = "\t", col.names = T, row.names = F) - }) + }) #download the selected rows only output$DownloadTableSelected <- downloadHandler( @@ -2420,10 +2523,10 @@ source = "volplot", priority = "event") proxy <- dataTableProxy("res_table") - + already_selected <- isolate(input$res_table_rows_selected) # proxy %>% selectRows(c(volplot_selected$key, already_selected)) - + #the following code is designed to allow deselection, but it is bugged for #some reason and requires the user to click the last selected node twice #to deselect it will need to work on this in later updates if required as @@ -2432,7 +2535,7 @@ # if(exists("volplot_selected") & !is.null(volplot_selected) & # length(already_selected) < 50){ if(length(already_selected) < 50){ - + if(length(intersect(volplot_selected$key, already_selected)) > 0){ #print("entering 'if' block") if(length(already_selected) > 1){ @@ -2458,12 +2561,12 @@ # print("entering 'else' block") # proxy %>% selectRows(NULL) # } - + } - + #volplot_selected <- NULL #print(isolate(global_data$trace_count)) - + }) @@ -2488,14 +2591,14 @@ list(format = format, width = width, height = height - ), + ), modeBarButtonsToRemove = list( "sendDataToCloud", "zoom2d", "zoomIn2d", "zoomOut2d", "pan2d", "select2d", "lasso2d", "autoScale2d", "hoverClosestCartesian", "hoverCompareCartesian" - ) ) + ) #updating the format for the counts plot plotlyProxy(outputId = "res_colplot", session = session) %>% @@ -2506,15 +2609,15 @@ list(format = format, width = width, height = height - ), + ), modeBarButtonsToRemove = list( "sendDataToCloud", "zoom2d", "zoomIn2d", "zoomOut2d", "pan2d", "select2d", "lasso2d", "autoScale2d", "toggleSpikelines"#, #"hoverClosestCartesian", #"hoverCompareCartesian" - ) ) + ) }) @@ -2579,7 +2682,7 @@ }) - # display TCR chord diagrams ---- + #display TCR chord diagrams observeEvent(input$TCRChordSelect, { output$TCRChordImage <- renderImage(expr = { @@ -2598,17 +2701,18 @@ width = dim(img)[2]*0.15, height = dim(img)[1]*0.15#, #alt = "This is alternate text" - ) + ) }, deleteFile = F) }) + tsne_aging_img <- function(filename){ img <- readPNG(source = filename, native = T, info = T) list( src = filename, contentType = 'image/png', - width = dim(img)[2]*0.5, - height = dim(img)[1]*0.5 + width = dim(img)[2]*0.15, + height = dim(img)[1]*0.15 ) } @@ -2617,23 +2721,26 @@ # aging tSNE plots ---- observeEvent(input$agingTSNEtissue, { tissue_files <- paste0("www/Aging_tsne_plots/", - aging_tsne_plots_list[grep(pattern = input$agingTSNEtissue, - x = aging_tsne_plots_list, - ignore.case = T)]) + aging_tsne_plots_list[grep(pattern = input$agingTSNEtissue, + x = aging_tsne_plots_list, + ignore.case = T)]) tissue_files <- tissue_files[c(6,2,3,4,5,1)] # ordering them manually output$agingTSNEImage1 <- renderImage( expr = tsne_aging_img(tissue_files[2]), deleteFile = F ) - + output$agingTSNEImage2 <- renderImage( expr = tsne_aging_img(tissue_files[2]), deleteFile = F ) + output$agingTSNETitle1 <- renderText(paste("t-SNE", input$agingTSNEtissue, "8 weeks")) + output$agingTSNETitle2 <- renderText(paste("t-SNE", input$agingTSNEtissue, "12 weeks")) + + }) - #display Pathview KEGG Pathways observeEvent(input$PathviewPlotSelect, { output$PathviewPlotImage <- renderImage(expr = { @@ -2646,14 +2753,14 @@ img <- readPNG( source = paste0("www/Pathview_plots/", Pathview_plots_list[grep(pattern = pathway_id, - x = Pathview_plots_list, - ignore.case = T)]), + x = Pathview_plots_list, + ignore.case = T)]), native = T, info = T) list(src = paste0("www/Pathview_plots/", Pathview_plots_list[grep(pattern = pathway_id, - x = Pathview_plots_list, - ignore.case = T)]), + x = Pathview_plots_list, + ignore.case = T)]), contentType = 'image/png', width = dim(img)[2],#*0.35, height = dim(img)[1]#*0.35#, @@ -2692,7 +2799,7 @@ output$DownloadPathviewSelected <- downloadHandler( filename = function() { paste0(input$PathviewPlotSelect, ".zip") - }, + }, #filename = paste0(input$PathviewPlotSelect, ".zip"), content = function(file){ @@ -2708,6 +2815,7 @@ ignore.case = T)]), "www/Legend.png", "www/Legend2.png"), flags = "-j") + }, contentType = "application/zip") # output$DownloadPathviewSelected <- downloadHandler( @@ -2732,7 +2840,7 @@ output$DownloadPathviewAll <- downloadHandler( filename = function() { paste0("all_pathways.zip") - }, + }, #filename = "all_pathways.zip", content = function(file){ @@ -2741,27 +2849,26 @@ "www/Legend.png", "www/Legend2.png"), flags = "-j") - }, + }, contentType = "application/zip") #download TCR plots - TCR_plots_list output$DownloadTCRSelected <- downloadHandler( filename = function() { paste0(input$TCRChordSelect, "_chord_diagram.png") - }, + }, #filename = paste0(input$TCRChordSelect, "_chord_diagram.png"), content = function(file){ file.copy(paste0("www/TCR_plots/", TCR_plots_list[grep(pattern = input$TCRChordSelect, - x = TCR_plots_list, - ignore.case = T)]), + x = TCR_plots_list, + ignore.case = T)]), file, copy.mode = F, copy.date = F - ) - }, + ) + }, contentType = "image/png") output$DownloadTCRAll <- downloadHandler( filename = function() { @@ -2799,7 +2906,7 @@ files = paste0("www/Global_bulk/", list.files(path = "www/Global_bulk/")), flags = "-j" - ) + ) }, contentType = "application/zip") #"GSEABulk", DiffExprsBulk @@ -2919,87 +3026,87 @@ "#WidthPixels", "#HeightPixels", "#res_volplot" - ), - intro = c(paste0("The results of the ", - "contrast are tabulated ", - "here. Column headers can ", - "be clicked to sort by ", - "that column (sorting can ", - "be done on multiple ", - "columns by pressing the ", - "'shift' key and clicking ", - "on the other header). ", - "Columns can be searched ", - "or filtered by clicking ", - "on the blank fields ", - "under the column headers.", - " Clicking a row will ", - "select the row in the ", - "table and highlight the ", - "respective gene in the ", - "volcano plot (upto 50 ", - "rows can be selected by ", - "pressing the 'shift' or ", - "'control' key)."), - paste0("Download the complete ", - "contrast results as a ", - "table."), - paste0("Download only the rows ", - "which are left after ", - "filtering (empty table if", - " no filtering done)."), - paste0("Download only the rows ", - "which have been manually ", - "selected (empty table if", - " no selection done)."), - paste0("This button resets any ", - "rows that have been ", - "selected."), - paste0("This checkbox shows and ", - "hides options related ", - "to exporting the plots."), - paste0("Select the format for ", - "exporting the plot."), - paste0("Specify the width of the ", - "plot in pixels for export."), - paste0("Specify the height of the ", - "plot in pixels for export."), - paste0("The interactive volcano ", - "plot of the contrast. ", - "Hovering over a data ", - "point would show a ", - "tooltip with additional ", - "information. Clicking on ", - "a data point will select ", - "the respective gene in ", - "the table and will also ", - "generate a column plot to ", - "visualise the expression ", - "of the selected gene in ", - "tissues. The plot can be ", - "zoomed-in by clicking and ", - "dragging a box over the ", - "area to be zoomed. ", - "Additionally the axes of ", - "the plot can be adjusted ", - "by dragging the edges of ", - "the respective axes. The ", - "zoom and/or axes can be ", - "reset by double-clicking ", - "in the empty space in the ", - "plot area or by clicking ", - "the house icon in the ", - "top-right of the plot. ", - "The plot can be exported ", - "using the export options ", - "by clicking the camera ", - "icon in the top-right of ", - "the plot. The same ", - "functionality except for ", - "data point selection also ", - "exists for the column ", - "plot.") - ))) + ), + intro = c(paste0("The results of the ", + "contrast are tabulated ", + "here. Column headers can ", + "be clicked to sort by ", + "that column (sorting can ", + "be done on multiple ", + "columns by pressing the ", + "'shift' key and clicking ", + "on the other header). ", + "Columns can be searched ", + "or filtered by clicking ", + "on the blank fields ", + "under the column headers.", + " Clicking a row will ", + "select the row in the ", + "table and highlight the ", + "respective gene in the ", + "volcano plot (upto 50 ", + "rows can be selected by ", + "pressing the 'shift' or ", + "'control' key)."), + paste0("Download the complete ", + "contrast results as a ", + "table."), + paste0("Download only the rows ", + "which are left after ", + "filtering (empty table if", + " no filtering done)."), + paste0("Download only the rows ", + "which have been manually ", + "selected (empty table if", + " no selection done)."), + paste0("This button resets any ", + "rows that have been ", + "selected."), + paste0("This checkbox shows and ", + "hides options related ", + "to exporting the plots."), + paste0("Select the format for ", + "exporting the plot."), + paste0("Specify the width of the ", + "plot in pixels for export."), + paste0("Specify the height of the ", + "plot in pixels for export."), + paste0("The interactive volcano ", + "plot of the contrast. ", + "Hovering over a data ", + "point would show a ", + "tooltip with additional ", + "information. Clicking on ", + "a data point will select ", + "the respective gene in ", + "the table and will also ", + "generate a column plot to ", + "visualise the expression ", + "of the selected gene in ", + "tissues. The plot can be ", + "zoomed-in by clicking and ", + "dragging a box over the ", + "area to be zoomed. ", + "Additionally the axes of ", + "the plot can be adjusted ", + "by dragging the edges of ", + "the respective axes. The ", + "zoom and/or axes can be ", + "reset by double-clicking ", + "in the empty space in the ", + "plot area or by clicking ", + "the house icon in the ", + "top-right of the plot. ", + "The plot can be exported ", + "using the export options ", + "by clicking the camera ", + "icon in the top-right of ", + "the plot. The same ", + "functionality except for ", + "data point selection also ", + "exists for the column ", + "plot.") + ))) observeEvent(input$TutorialButton2,{ updateCheckboxInput(session = session, From 8539983be527a255c71349961ecb602f7b02de5d Mon Sep 17 00:00:00 2001 From: Laura Biggins Date: Tue, 29 Aug 2023 11:12:08 +0100 Subject: [PATCH 04/15] Updated the tSNE plots to the shiny server version --- expressionViewer/data/tSNE_plots.rds | Bin 3017658 -> 3126543 bytes 1 file changed, 0 insertions(+), 0 deletions(-) diff --git a/expressionViewer/data/tSNE_plots.rds b/expressionViewer/data/tSNE_plots.rds index 81e2f3ead801e4ece3c61f143f81b4ffaec02012..9e97ddc7e9ec8fb4165288518a8acff1cbea5e9d 100644 GIT binary patch literal 3126543 zcmV){Kz+X-iwFP!000002JF2FcpS%ZDBOb^Byo}gZ*mDz6iJZ~PhAu#NE9Veq9}@@ zBgubyy1M5T3xp8OqNS->G`HISiq^Z1 z-?;5c`=7;5glLw}PIxxi&~3w}b9Zddcb@vmZ=JjSzxH&lTlLy=cijE+`&)ndyXWrs z?Y}6F{p~ZucVD%C_@d7hh97+6>ARQx4=rqZ-Lk2F z86Nqq=el=Xv3+>tbN|}%6Uz>qJM??ke&h7F4xKyvSF#e>`{i 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