inst/ProstarApp/server/srv_AnaDiff.R

source(file.path("server", "mod_popover.R"), local = TRUE)$value
source(file.path("server", "mod_query_metacell.R"), local = TRUE)$value
source(file.path("server", "mod_filtering_example.R"), local = TRUE)$value
source(file.path("server", "mod_staticDT.R"), local=TRUE)$value


convertAnaDiff2DF <- reactive({
  req(rv$widgets$anaDiff)
  rv$widgets$anaDiff[sapply(rv$widgets$anaDiff, is.null)] <- NA
  df <- as.data.frame(tibble::enframe(rv$widgets$anaDiff))
  names(df) <- c("Parameter", "Value")
  rownames(df) <- NULL
  df
})


callModule(moduleVolcanoplot,"volcano_Step1", 
           data = reactive({rv$resAnaDiff}),
           comp = reactive({as.character(rv$widgets$anaDiff$Comparison)}),
           tooltip = reactive({ rv$widgets$anaDiff$tooltipInfo})
           )

callModule(moduleVolcanoplot,"volcano_Step2",
           data = reactive({rv$resAnaDiff}),
           comp = reactive({as.character(rv$widgets$anaDiff$Comparison)}),
           tooltip = reactive({rv$widgets$anaDiff$tooltipInfo})
           )

mod_staticDT_server("params_AnaDiff",
             data = reactive({convertAnaDiff2DF()}),
             filename='AnaDiffParams')

callModule(module_Not_a_numeric,"test_seuilPVal", reactive({rv$widgets$anaDiff$th_pval}))


callModule(moduleProcess, "moduleProcess_AnaDiff", 
           isDone = reactive({rvModProcess$moduleAnaDiffDone}), 
           pages = reactive({rvModProcess$moduleAnaDiff}),
           rstFunc = resetModuleAnaDiff,
           forceReset = reactive({rvModProcess$moduleAnaDiffForceReset}))


######
resetModuleAnaDiff <- reactive({  
  # if (rv$widgets$anaDiff$swapVolcano == TRUE){
  #   rv$resAnaDiff$logFC <- - rv$resAnaDiff$logFC
  #   rv$widgets$anaDiff$swapVolcano <- FALSE
  # }

  ## update widgets values (reactive values)
  resetModuleProcess("AnaDiff")
  
  rv$nbTotalAnaDiff = NULL
  rv$nbSelectedAnaDiff = NULL
  rv$nbSelectedTotal_Step3 = NULL
  rv$nbSelected_Step3 = NULL  
  rv$conditions <- list(cond1 = NULL, cond2 = NULL)
  rv$calibrationRes <- NULL
  rv$errMsgcalibrationPlot <- NULL
  rv$errMsgcalibrationPlotALL <- NULL
  rv$pi0 <- NULL
  
  rv$widgets$anaDiff$Comparison = "None"
  rv$widgets$anaDiff$Condition1 = ""
  rv$widgets$anaDiff$Condition2 = ""
  rv$widgets$anaDiff$val_vs_percent = "Value"
  rv$widgets$anaDiff$ChooseFilters = "None"
  rv$widgets$anaDiff$seuilNA_percent = 0
  rv$widgets$anaDiff$seuilNA = 0
  rv$widgets$anaDiff$filter_th_NA = 0
  rv$widgets$anaDiff$calibMethod = 'None'
  rv$widgets$anaDiff$numValCalibMethod = 0
  rv$widgets$anaDiff$th_pval = 0
  rv$widgets$anaDiff$FDR = 0
  rv$widgets$anaDiff$NbSelected = 0
  rv$widgets$anaDiff$nBinsHistpval = 80
  rv$widgets$anaDiff$downloadAnaDiff = "All"
  rv$widgets$anaDiff$tooltipInfo = rv$current.obj@experimentData@other$proteinId
  
  rv$widgets$anaDiff[sapply(rv$widgets$anaDiff, is.null)] <- NA
  rvModProcess$moduleAnaDiffDone = rep(FALSE, 4)
  
  rv_anaDiff$filename = NULL
  rv_anaDiff$local.reset <- rv_anaDiff$local.reset + 1
  UpdateCompList()
  ##update dataset to put the previous one
  #rv$current.obj <- rv$dataset[[last(names(rv$dataset))]] 
  #rv$resAnaDiff <- NULL
  
  
})


rv_anaDiff <- reactiveValues(
  filename = NULL,
  local.reset = 0
)




###
### ------------------- SCREEN 1 ------------------------------
###


# Fill the variable 'rv$resAnaDiff' with informations relatives to
# the comparison choosen by the user. 
# Concertely, it extracts data from the variable rv$res_AllPairwiseComparisons
# which contains all info (logFC and pValue) for all comparisons.
UpdateCompList <- reactive({
  rv$widgets$anaDiff$Comparison 
  isolate({
    if (rv$widgets$anaDiff$Comparison== "None"){
      rv$resAnaDiff <- NULL
      return(NULL)
    } else {
      index <- which(paste(as.character(rv$widgets$anaDiff$Comparison), "_logFC", sep="") == colnames(rv$res_AllPairwiseComparisons$logFC))
      
      # Update of the list rv$resAnaDiff
      rv$widgets$anaDiff$Condition1 <- strsplit(as.character(rv$widgets$anaDiff$Comparison), "_vs_")[[1]][1]
      rv$widgets$anaDiff$Condition2 <- strsplit(as.character(rv$widgets$anaDiff$Comparison), "_vs_")[[1]][2]
      
      # if (input$swapVolcano == FALSE)
        rv$resAnaDiff <- list(logFC = (rv$res_AllPairwiseComparisons$logFC)[,index],
                            P_Value = (rv$res_AllPairwiseComparisons$P_Value)[,index],
                            condition1 = rv$widgets$anaDiff$Condition1,
                            condition2 = rv$widgets$anaDiff$Condition2
      )
      # # else
      #   rv$resAnaDiff <- list(logFC = (rv$res_AllPairwiseComparisons$logFC)[,index],
      #                         P_Value = -(rv$res_AllPairwiseComparisons$P_Value)[,index],
      #                         condition1 = rv$widgets$anaDiff$Condition1,
      #                         condition2 = rv$widgets$anaDiff$Condition2
      #   )
      
      rvModProcess$moduleAnaDiffDone[1] <- TRUE
    }
  })
})


# By default, the tooltip for volcanoplot is set to the proteinId
observe({
  req(rv$current.obj)
  if (is.null(rv$widgets$anaDiff$tooltipInfo))
    rv$widgets$anaDiff$tooltipInfo <- rv$current.obj@experimentData@other$proteinId
  
})


output$screenAnaDiff1 <- renderUI({
  isolate({
    tagList(
      tags$div(
        tags$div( style="display:inline-block; vertical-align: top; padding-right: 60px",
                  selectInput("selectComparison", "Select a comparison",
                                choices = c("None" = "None", GetPairwiseCompChoice()),
                                selected = rv$widgets$anaDiff$Comparison,
                                width = '300px')
                  )
         ),
        uiOutput('pushpval_ui'),

      tags$hr(),
      tags$div(
        tags$div( style="display:inline-block; vertical-align: top; padding-right: 60px",
                  moduleVolcanoplotUI("volcano_Step1")),
        tags$div( style="display:inline-block; vertical-align: top;",
                  tagList(
                    br(),
                    uiOutput("volcanoTooltip_UI")
                    )
                  )
        )
    )
    
  })
})


output$pushpval_ui <- renderUI({
  req(rv$widgets$anaDiff$Comparison != 'None')
  callModule(modulePopover, "modulePopover_pushPVal", 
             data = reactive(list(title=h3("Push p-value"),
                                  content= "This functionality is useful in case of multiple pairwise comparisons (more than 2 conditions): At the filtering step, a given analyte X (either peptide or protein) may have been kept because it contains very few missing values in a given condition (say Cond. A), even though it contains (too) many of them in all other conditions (say Cond B and C only contains “MEC” type missing values). Thanks to the imputation step, these missing values are no longer an issue for the differential analysis, at least from the computational viewpoint. However, statistically speaking, when performing B vs C, the test will rely on too many imputed missing values to derive a meaningful p-value: It may be wiser to consider analyte X as non-differentially abundant, regardless the test result (and thus, to push its p-value to 1). This is just the role of the “P-value push” parameter. It makes it possible to introduce a new filtering step that only applies to each pairwise comparison, and which assigns a p-value of 1 to analytes that, for the considered comparison are assumed meaningless due to too many missing values (before imputation).")))
  
    wellPanel(
       modulePopoverUI("modulePopover_pushPVal"),
      mod_query_metacell_ui('AnaDiff_query'),
      shinyjs::disabled(actionButton("AnaDiff_performFilteringMV",
                                   "Push p-value",
                                   class = actionBtnClass)
                      )
    )
})

#---------------------------

# Extract conditions of the current dataset to represent the selected comparison
# It returns a subset of the current dataset that will be used to filter the data
# within the 'Push p-value' feature
Get_Dataset_to_Analyze <- reactive({
  req(rv$widgets$anaDiff$Comparison != 'None')
  req(rv$current.obj)

  datasetToAnalyze <- NULL
  
  if (length(grep("all-", rv$widgets$anaDiff$Comparison)) == 1){
    condition1 = strsplit(as.character(rv$widgets$anaDiff$Comparison), "_vs_")[[1]][1]
    ind_virtual_cond2 <- which(pData(rv$current.obj)$Condition != condition1)
    datasetToAnalyze <- rv$current.obj
    pData(datasetToAnalyze)$Condition[ind_virtual_cond2] <- 'virtual_cond_2'
  }
  else {
    condition1 = strsplit(as.character(rv$widgets$anaDiff$Comparison), "_vs_")[[1]][1]
    condition2 = strsplit(as.character(rv$widgets$anaDiff$Comparison), "_vs_")[[1]][2]
    
    if (substr(condition1, 1,1) == '(' && 
        substr(condition1, nchar(condition1),nchar(condition1)) == ')')
      condition1 <- sub('^.(.*).$', '\\1', condition1)
    
    if (substr(condition2, 1,1) == '(' && 
        substr(condition2, nchar(condition2), nchar(condition2)) == ')')
      condition2 <- sub('^.(.*).$', '\\1', condition2)
    
    
    
    ind <- c( which(pData(rv$current.obj)$Condition == condition1), 
              which(pData(rv$current.obj)$Condition == condition2))
    
    datasetToAnalyze <- rv$current.obj[,ind]
    datasetToAnalyze@experimentData@other$names_metacell <-
      rv$current.obj@experimentData@other$names_metacell[ind]
  }
  
  datasetToAnalyze
}) %>% bindCache(rv$current.obj, rv$widgets$anaDiff$Comparison)






AnaDiff_indices <- mod_query_metacell_server(id = 'AnaDiff_query',
                                     obj = reactive({Get_Dataset_to_Analyze()}),
                                     list_tags = reactive({c('None' = 'None',
                                                             DAPAR::metacell.def(GetTypeofData(rv$current.obj))$node
                                                             )
                                       }),
                                     keep_vs_remove = reactive({setNames(nm = c("delete", "keep"))}),
                                     filters = reactive({c("None" = "None",
                                                           "Whole Line" = "WholeLine",
                                                           "Whole matrix" = "WholeMatrix",
                                                           "For every condition" = "AllCond",
                                                           "At least one condition" = "AtLeastOneCond")}),
                                     val_vs_percent = reactive({setNames(nm=c('Count', 'Percentage'))}),
                                     operator = reactive({setNames(nm=DAPAR::SymFilteringOperators())}),
                                     reset = reactive({rv_anaDiff$local.reset})
                                     )
#----------------------------------------------------



observe({
  req(AnaDiff_indices()$params$MetacellTag)

  shinyjs::toggleState("AnaDiff_performFilteringMV",
                       condition = AnaDiff_indices()$params$MetacellTag != 'None')
})

########################################################
## Perform missing values filtering
########################################################
observeEvent(input$AnaDiff_performFilteringMV, ignoreInit = TRUE, ignoreNULL = TRUE, {
   UpdateCompList()
  rv$widgets$anaDiff$MetacellTag <- AnaDiff_indices()$params$MetacellTag
  rv$widgets$anaDiff$KeepRemove <-  AnaDiff_indices()$params$KeepRemove
  rv$widgets$anaDiff$ChooseFilters <- AnaDiff_indices()$params$MetacellFilters
  rv$widgets$anaDiff$seuilNA_percent  <- AnaDiff_indices()$params$metacell_percent_th
  rv$widgets$anaDiff$seuilNA  <- AnaDiff_indices()$params$metacell_value_th
  rv$widgets$anaDiff$val_vs_percent  <- AnaDiff_indices()$params$val_vs_percent
  rv$widgets$anaDiff$operator  <- AnaDiff_indices()$params$metacellFilter_operator
  rv$widgets$anaDiff$tooltipInfo <- rv$current.obj@experimentData@other$proteinId
  
  if (as.character(rv$widgets$anaDiff$ChooseFilters) == 'None')
    GetBackToCurrentResAnaDiff()

    #--------------------------------
    if (!is.null(AnaDiff_indices()$indices) && length(AnaDiff_indices()$indices) < nrow(Get_Dataset_to_Analyze())){
      rv$resAnaDiff$P_Value[-AnaDiff_indices()$indices] <- 1
    }

})



# Catch a change with swapVolcano, made by the user
# observeEvent(req(input$swapVolcano),{
#   req(rv$resAnaDiff$logFC)
# 
#   rv$widgets$anaDiff$swapVolcano <- input$swapVolcano
#   rv$resAnaDiff$logFC <- -rv$resAnaDiff$logFC
# 
# })


observeEvent(input$selectComparison,ignoreInit = TRUE,{ 
  rv$widgets$anaDiff$Comparison <- input$selectComparison
  rv$widgets$anaDiff$tooltipInfo <- rv$current.obj@experimentData@other$proteinId
  UpdateCompList()
  
  req(rv$widgets$anaDiff$Comparison)
  cond1 = rv$widgets$anaDiff$Condition1
  cond2 = rv$widgets$anaDiff$Condition2
  
  # if (isTRUE(rv$widgets$anaDiff$swapVolcano)) {
  #   rv_anaDiff$filename = paste0('anaDiff_', cond2,'_vs_', cond1, '.xlsx')
  # } else {
    rv_anaDiff$filename = paste0('anaDiff_', cond1,'_vs_', cond2, '.xlsx')
  #}
  
})





##--------------------------------------------------------
##---------------------------------------------------------


output$volcanoTooltip_UI <- renderUI({
  req(rv$widgets$anaDiff$Comparison != "None")
  rv$widgets$anaDiff$tooltipInfo
  isolate({
    tagList(
      modulePopoverUI("modulePopover_volcanoTooltip"),
      selectInput("tooltipInfo",
                  label = NULL,
                  choices = colnames(fData(rv$current.obj)),
                  selected = rv$widgets$anaDiff$tooltipInfo,
                  multiple = TRUE,
                  selectize = FALSE,
                  width='300px', size=5),
      actionButton("validTooltipInfo", "Validate tooltip choice", class = actionBtnClass)
    )
  })
})



GetPairwiseCompChoice <- reactive({
  req(rv$res_AllPairwiseComparisons$logFC)
  ll <- unlist(strsplit(colnames(rv$res_AllPairwiseComparisons$logFC), "_logFC"))
  ll
})




# observe({
#   req(rv$widgets$anaDiff$Comparison)
#   #shinyjs::toggle('toto', condition = rv$widgets$anaDiff$Comparison != "None")
#   shinyjs::toggle('swapVolcano', condition = rv$widgets$anaDiff$Comparison != "None")
# })


callModule(modulePopover,"modulePopover_volcanoTooltip", 
           data = reactive(list(title = "Tooltip", 
                                content="Infos to be displayed in the tooltip of volcanoplot")))


callModule(modulePopover,"modulePopover_keepLines", 
           data = reactive(list(title="n values",
                                content= "Keep the lines which have at least n intensity values.")))


GetBackToCurrentResAnaDiff <- reactive({
  req(rv$res_AllPairwiseComparisons)
  
  index <- which(paste(as.character(rv$widgets$anaDiff$Comparison), "_logFC", sep="") == colnames(rv$res_AllPairwiseComparisons$logFC))
  rv$resAnaDiff <- list(logFC = (rv$res_AllPairwiseComparisons$logFC)[,index],
                        P_Value = (rv$res_AllPairwiseComparisons$P_Value)[,index],
                        condition1 = strsplit(as.character(rv$widgets$anaDiff$Comparison), "_vs_")[[1]][1],
                        condition2 = strsplit(as.character(rv$widgets$anaDiff$Comparison), "_vs_")[[1]][2]
  )
  rv$resAnaDiff
})





not_a_numeric <- function(input) {
  if (is.na(as.numeric(input))) {
    "Please input a number"
  }  else {
    NULL
  }
}

# 
# Get_seuilPVal <- reactive({
#    shiny::validate(
#     need(!is.na(rv$widgets$anaDiff$th_pval), "")
#   )
#   rv$widgets$anaDiff$th_pval
# })



###
### ------------------- SCREEN 2 ------------------------------
###



output$screenAnaDiff2 <- renderUI({
  req(as.character(rv$widgets$anaDiff$Comparison))
  
  tagList(
    tags$div(
      tags$div( style="display:inline-block; vertical-align: middle; padding-right: 40px;",
                selectInput("calibrationMethod","Calibration method",
                            choices = c('None' = 'None', calibMethod_Choices),
                            selected = rv$widgets$anaDiff$calibMethod, width='200px')
      ),
      tags$div( style="display:inline-block; vertical-align: middle;",
                uiOutput('numericalValForCalibrationPlot')
                #hidden(numericInput( "numericValCalibration","Proportion of TRUE null hypohtesis", 
                #                     value = rv$widgets$anaDiff$numValCalibMethod, 
                #                     min=0, max=1, step=0.05, width='200px')
                #      )
      ),
      tags$div( style="display:inline-block; vertical-align: middle;padding-right: 40px;",
                uiOutput('nBins_ui')),
      tags$div( style="display:inline-block; vertical-align: middle;",
                p(tags$strong(paste0("value of pi0: ", round(as.numeric(rv$pi0), digits=2))))
      )
      
    ),
    tags$hr(),
    
    fluidRow(
      column(width=6,fluidRow(style = "height:800px;",imageOutput("calibrationPlotAll", height='800px'))),
      column(width=6,fluidRow(style = "height:400px;",imageOutput("calibrationPlot", height='400px')),
             fluidRow(style = "height:400px;",highchartOutput("histPValue"))
      )
    )
  )
  
})    





observeEvent(input$nBinsHistpval,{ 
  rv$widgets$anaDiff$nBinsHistpval <- as.numeric(input$nBinsHistpval)
  })


observeEvent(input$calibrationMethod,{  
  rv$widgets$anaDiff$calibMethod <- input$calibrationMethod
  shinyjs::toggle("numericValCalibration", condition=rv$widgets$anaDiff$calibMethod == "numeric value")
})


observeEvent(input$numericValCalibration,{  
  rv$widgets$anaDiff$numValCalibMethod <- input$numericValCalibration
})


output$nBins_ui <- renderUI({
  req(rv$resAnaDiff)
  req(rv$pi0)
  req(rv$widgets$anaDiff$nBinsHistpval)
  
  selectInput("nBinsHistpval", "n bins of p-value histogram", 
              choices = c(1,seq(from = 0, to = 100, by = 10)[-1]),
              selected=rv$widgets$anaDiff$nBinsHistpval, width='80px')
})


histPValue <- reactive({

  req(rv$resAnaDiff)
  req(rv$pi0)
  req(rv$widgets$anaDiff$nBinsHistpval)
  
  rv$widgets$hypothesisTest$th_logFC
  
  if (is.null(rv$widgets$hypothesisTest$th_logFC) || is.na(rv$widgets$hypothesisTest$th_logFC) ||
      (length(rv$resAnaDiff$logFC) == 0)) { return()}
  m <- match.metacell(DAPAR::GetMetacell(rv$current.obj), 
                      pattern="missing", 
                      level = 'peptide')
  if (length(which(m)) > 0)
    {return()}
  
  # isolate({
  t <- NULL
  method <- NULL
  t <- rv$resAnaDiff$P_Value
  t <- t[which(abs(rv$resAnaDiff$logFC) >= rv$widgets$hypothesisTest$th_logFC)]
  toDelete <- which(t==1)
  if (length(toDelete) > 0){	t <- t[-toDelete] }
  histPValue_HC(t,
                bins=as.numeric(rv$widgets$anaDiff$nBinsHistpval), 
                pi0=rv$pi0)
  # })
})

output$histPValue <- renderHighchart({
  histPValue()
})



output$numericalValForCalibrationPlot <- renderUI({
  rv$widgets$anaDiff$calibMethod
  if (rv$widgets$anaDiff$calibMethod == "numeric value"){
    numericInput( "numericValCalibration","Proportion of TRUE null hypohtesis",
                  value = rv$widgets$anaDiff$numValCalibMethod,
                  min=0, max=1, step=0.05)
  }
})


output$calibrationResults <- renderUI({
  req(rv$calibrationRes)
  rv$widgets$hypothesisTest$th_logFC
  rv$current.obj
  
  
  txt <- paste("Non-DA protein proportion = ", 
               round(100*rv$calibrationRes$pi0, digits = 2),"%<br>",
               "DA protein concentration = ", 
               round(100*rv$calibrationRes$h1.concentration, digits = 2),
               "%<br>",
               "Uniformity underestimation = ", 
               rv$calibrationRes$unif.under,"<br><br><hr>", sep="")
  HTML(txt)
  
})




calibrationPlot <- reactive({
  req(rv$widgets$anaDiff$calibMethod != 'None')
  rv$widgets$hypothesisTest$th_logFC
  rv$resAnaDiff
  req(rv$current.obj)
  
  if (length(rv$resAnaDiff$logFC) == 0) { return()}
  
  m <- match.metacell(DAPAR::GetMetacell(rv$current.obj), 
                      pattern="missing", 
                      level = 'peptide')
  if (length(which(m)) > 0)
    return()
  
  cond <- c(rv$resAnaDiff$condition1, rv$resAnaDiff$condition2)
  
  
  t <- NULL
  method <- NULL
  t <- rv$resAnaDiff$P_Value
  t <- t[which(abs(rv$resAnaDiff$logFC) >= rv$widgets$hypothesisTest$th_logFC)]
  toDelete <- which(t==1)
  if (length(toDelete) > 0){	t <- t[-toDelete] }
  
  l <- NULL
  ll <- NULL
  result = tryCatch(
    {
      
      if ((rv$widgets$anaDiff$calibMethod == "numeric value") 
          && !is.null(rv$widgets$anaDiff$numValCalibMethod)) {
        
        ll <-catchToList(
          wrapperCalibrationPlot(t, 
                                 rv$widgets$anaDiff$numValCalibMethod))
        rv$errMsgCalibrationPlot <- ll$warnings[grep( "Warning:", ll$warnings)]
      }
      else if (rv$widgets$anaDiff$calibMethod == "Benjamini-Hochberg") {
        
        ll <-catchToList(wrapperCalibrationPlot(t, 1))
        rv$errMsgCalibrationPlot <- ll$warnings[grep( "Warning:", ll$warnings)]
      }else { 
        ll <-catchToList(wrapperCalibrationPlot(t, rv$widgets$anaDiff$calibMethod))
        rv$errMsgCalibrationPlot <- ll$warnings[grep( "Warning:", ll$warnings)]
      }
      rv$pi0 <- ll$value$pi0

      rvModProcess$moduleAnaDiffDone[2] <- !is.null(rv$pi0)
    }
    , warning = function(w) {
      shinyjs::info(paste("Calibration plot",":",
                          conditionMessage(w), sep=" "))
    }, error = function(e) {
      shinyjs::info(paste("Calibration plot",":",
                          conditionMessage(e), sep=" "))
    }, finally = {
      #cleanup-code 
    })

  
})

output$calibrationPlot <- renderImage({
  
  outfile <- tempfile(fileext='.png')
  
  # Generate a png
  png(outfile, width=600, height=500)
  calibrationPlot()
  dev.off()
  
  # Return a list
  list(src = outfile,
       alt = "This is alternate text")
}, deleteFile = TRUE)




output$errMsgCalibrationPlot <- renderUI({
  req(rv$errMsgCalibrationPlot)
  rv$widgets$hypothesisTest$th_logFC
  req(rv$current.obj)
  
  txt <- NULL
  
  for (i in 1:length(rv$errMsgCalibrationPlot)) {
    txt <- paste(txt, "errMsgCalibrationPlot: ",rv$errMsgCalibrationPlot[i], "<br>", sep="")
  }
  
  div(HTML(txt), style="color:red")
  
})


output$errMsgCalibrationPlotAll <- renderUI({
  rv$errMsgCalibrationPlotAll
  rv$widgets$hypothesisTest$th_logFC
  req(rv$current.obj)
  if (is.null(rv$errMsgCalibrationPlotAll) ) {return()}
  
  txt <- NULL
  for (i in 1:length(rv$errMsgCalibrationPlotAll)) {
    txt <- paste(txt, "errMsgCalibrationPlotAll:", rv$errMsgCalibrationPlotAll[i], "<br>", sep="")
  }
  
  div(HTML(txt), style="color:red")
})



calibrationPlotAll <- reactive({
  
  rv$resAnaDiff
  req(rv$current.obj)
  
  if (is.na(rv$widgets$hypothesisTest$th_logFC) ||
      (length(rv$resAnaDiff$logFC) == 0)) { return()}
  m <- match.metacell(DAPAR::GetMetacell(rv$current.obj), 
                      pattern="missing", 
                      level = 'peptide')
  if (length(which(m)) > 0)
    return()
  
  cond <- c(rv$resAnaDiff$condition1, rv$resAnaDiff$condition2)

  t <- NULL
  method <- NULL
  t <- rv$resAnaDiff$P_Value
  t <- t[which(abs(rv$resAnaDiff$logFC) >= rv$widgets$hypothesisTest$th_logFC)]
  toDelete <- which(t==1)
  if (length(toDelete) > 0){
    t <- t[-toDelete]
  }
  
  l <- NULL
  result = tryCatch(
    {
      l <-catchToList(wrapperCalibrationPlot(t, "ALL")  )
      rv$errMsgCalibrationPlotAll <- l$warnings[grep( "Warning:",l$warnings)]
      rvModProcess$moduleAnaDiffDone[2] <- !is.null(rv$pi0)
    }
    , warning = function(w) {
      shinyjs::info(paste("Calibration Plot All methods",":",
                          conditionMessage(w), sep=" "))
    }, error = function(e) {
      shinyjs::info(paste("Calibration Plot All methods",":",
                          conditionMessage(e), sep=" "))
    }, finally = {
      #cleanup-code 
    })
  
})



#--------------------------------------------------
output$calibrationPlotAll <- renderImage({
  
  outfile <- tempfile(fileext='.png')
  
  # Generate a png
  png(outfile, width=600, height=500)
  calibrationPlotAll()
  dev.off()
  
  # Return a list
  list(src = outfile,
       alt = "This is alternate text")
}, deleteFile = TRUE)



###
### ------------------- SCREEN 3 ------------------------------
###

output$screenAnaDiff3 <- renderUI({
  print("in output$screenAnaDiff3")
  
  if(as.character(rv$widgets$anaDiff$Comparison) == "None"){return(NULL)}
  
  isolate({
    tagList(
      tags$div(
        tags$div( style="display:inline-block; vertical-align: center; padding-right: 2px;",
                  modulePopoverUI("modulePopover_pValThreshold"),
                  textInput("seuilPVal",  NULL,
                            value=rv$widgets$anaDiff$th_pval, width='100px')),
        actionButton("valid_seuilPVal", 'Validate value', class = actionBtnClass),
        tags$div( style="display:inline-block; vertical-align: top;",
                  module_Not_a_numericUI("test_seuilPVal"))
        
        
      ),
      tags$hr(),
      tagList(
        tags$div(
          tags$div( style="display:inline-block; vertical-align: top;",
                    htmlOutput("showFDR"),
                    withProgress(message = '',detail = '', value = 1, {
                      moduleVolcanoplotUI("volcano_Step2")
                    })
          ),
          
          tags$div( style="display:inline-block; vertical-align: top;",
                    uiOutput("tooltipInfo"),
                    checkboxInput("showpvalTable","Show p-value table", value=FALSE),
                    radioButtons("downloadAnaDiff", "Download as Excel file", 
                                 choices=c("All data"="All", "only DA"="onlyDA" ),
                                 selected = rv$widgets$anaDiff$downloadAnaDiff),
                    downloadButton('downloadSelectedItems', 'Download', class=actionBtnClass))
        ),
        hidden(DTOutput("anaDiff_selectedItems"))
      )
    )
    
  })
})     



###
### ------------------- SCREEN 4 ------------------------------
###



output$screenAnaDiff4 <- renderUI({     
  req(as.character(rv$widgets$anaDiff$Comparison) != "None")
  tagList(
    mod_static_ui("params_AnaDiff")
    
  )
})

output$diffAna_Summary <- renderUI({     
  req(as.character(rv$widgets$anaDiff$Comparison) != "None")
  
  tagList(
    mod_static_ui("params_AnaDiff")
  )
  
})


##################################################################################
###### Set code for widgets managment
##################################################################################







observeEvent(input$valid_seuilPVal,{ 
  req(input$seuilPVal)
  tmp <- gsub(",", ".", input$seuilPVal, fixed=TRUE)
  
  rv$widgets$anaDiff$th_pval <- as.numeric(tmp)
})



observeEvent(input$showpvalTable, {
  print("show : anaDiff_selectedItems")
  shinyjs::toggle(id = "anaDiff_selectedItems", condition=isTRUE(input$showpvalTable))
})


observeEvent(input$validTooltipInfo,{ 
  rv$widgets$anaDiff$tooltipInfo <- unique(rv$current.obj@experimentData@other$proteinid, input$tooltipInfo)
  })

observeEvent(input$downloadAnaDiff,{  rv$widgets$anaDiff$downloadAnaDiff <- input$downloadAnaDiff})

callModule(modulePopover,"modulePopover_pValThreshold", 
           data = reactive(list(title = "p-val cutoff", 
                                content="Define the -log10(p_value) threshold")))


output$anaDiff_selectedItems <- renderDT({
  
  DT::datatable(GetSelectedItems(),
                escape = FALSE,
                rownames=FALSE,
                options = list(
                  initComplete = initComplete(),
                  dom = 'frtip',
                  server = TRUE,
                  columnDefs = list(list(width='200px',targets= "_all")),
                  ordering = TRUE)
  ) %>%
    formatStyle(
      paste0('isDifferential (', as.character(rv$widgets$anaDiff$Comparison), ')'),
      target = 'row',
      backgroundColor = styleEqual(c(0, 1), c("white", orangeProstar))
    )
})



output$downloadSelectedItems <- downloadHandler(
  filename = reactive({rv_anaDiff$filename}),
  
  content = function(file) {
    DA_Style <- openxlsx::createStyle(fgFill = orangeProstar)
    hs1 <- openxlsx::createStyle(fgFill = "#DCE6F1", halign = "CENTER", textDecoration = "italic",
                                 border = "Bottom")
    wb <- openxlsx::createWorkbook() # Create wb in R
    openxlsx::addWorksheet(wb,sheetName="DA result") #create sheet
    openxlsx::writeData(wb,sheet = 1, as.character(rv$widgets$anaDiff$Comparison), colNames = TRUE,headerStyle = hs1)
    openxlsx::writeData(wb,sheet = 1, startRow = 3, GetSelectedItems(), colNames = TRUE)
    ll.DA.row <- which(GetSelectedItems()[,paste0('isDifferential (',as.character(rv$widgets$anaDiff$Comparison), ')')]==1)
    ll.DA.col <- rep(which(colnames(GetSelectedItems()) == paste0('isDifferential (',as.character(rv$widgets$anaDiff$Comparison),')')), length(ll.DA.row))
    
    openxlsx::addStyle(wb, sheet=1, cols=ll.DA.col,
                       rows = 3 + ll.DA.row, style = DA_Style)
    
    tempFile <- tempfile(fileext = ".xlsx")
    openxlsx::saveWorkbook(wb, file = tempFile, overwrite = TRUE)
    
    file.rename(tempFile, file)
    
  })











# output$anaDiff_selectedItems <- renderDT({
#   
#   DT::datatable(GetSelectedItems(),
#                 escape = FALSE,
#                 rownames=TRUE,
#                 options = list(initComplete = initComplete(),
#                                dom = 'Bfrtip',
#                                server = TRUE,
#                                columnDefs = list(list(width='200px',targets= "_all")),
#                                ordering = TRUE)
#   ) %>%
#     formatStyle(
#       'isDifferential',
#       target = 'row',
#       backgroundColor = styleEqual(c(0, 1), c("white",orangeProstar))
#     )
# })



# output$downloadSelectedItems <- downloadHandler(
#   #input$chooseDatasetToExportToMSnset,
#   filename = paste0('diffanalysis_', input$datasets,'.xlsx'),
#   content = function(file) {
#     print(paste0("file to write=", file))
#     DA_Style <- openxlsx::createStyle(fgFill = orangeProstar)
#     hs1 <- createStyle(fgFill = "#DCE6F1", halign = "CENTER", textDecoration = "italic",
#                        border = "Bottom")
#     wb <- openxlsx::createWorkbook() # Create wb in R
#     openxlsx::addWorksheet(wb,sheetName="DA result") #create sheet
#     openxlsx::writeData(wb,sheet = 1, as.character(rv$widgets$anaDiff$Comparison), colNames = TRUE,headerStyle = hs1)
#     openxlsx::writeData(wb,sheet = 1, startRow = 3,GetSelectedItems(), colNames = TRUE)
#     ll.DA.row <- which(GetSelectedItems()[,'isDifferential']==1)
#     ll.DA.col <- rep(which(colnames(GetSelectedItems()) == 'isDifferential'), length(ll.DA.row))
#     openxlsx::addStyle(wb, sheet=1, cols=ll.DA.col, rows = 1+ ll.DA.row, style = DA_Style)
#     openxlsx::saveWorkbook(wb, file = file, overwrite = TRUE)
#   })
# 













#####
####  SELECT AND LOAD ONE PARIWISE COMPARISON
####







Get_FDR <- reactive({
  req(rv$current.obj)
  rv$widgets$anaDiff$numValCalibMethod
  rv$widgets$anaDiff$calibMethod
  req(rv$resAnaDiff)
  
  m <- NULL
  if (rv$widgets$anaDiff$calibMethod == "Benjamini-Hochberg") { m <- 1}
  else if (rv$widgets$anaDiff$calibMethod == "numeric value") {
    m <- as.numeric(rv$widgets$anaDiff$numValCalibMethod)} 
  else {m <- rv$widgets$anaDiff$calibMethod }
  
  rv$widgets$anaDiff$FDR <- diffAnaComputeFDR(rv$resAnaDiff[["logFC"]], 
                                              rv$resAnaDiff[["P_Value"]],
                                              rv$widgets$anaDiff$th_pval, 
                                              rv$widgets$hypothesisTest$th_logFC, 
                                              m)
  rvModProcess$moduleAnaDiffDone[3] <- TRUE
  as.numeric(rv$widgets$anaDiff$FDR)
})


output$showFDR <- renderUI({
  req(rv$current.obj)
  nb <- length(which(GetSelectedItems()[paste0('isDifferential (',as.character(rv$widgets$anaDiff$Comparison),')')]==1))
  th <- Get_FDR() * nb
  print(th)
  
  
  tagList(
    if (!is.infinite(Get_FDR())){
      tags$p(style="font-size: 25px;","FDR = ", round(100*Get_FDR(), digits=2)," % (p-value = ",
             signif(10^(- (rv$widgets$anaDiff$th_pval)), digits=3), ")")
    } else {
      tags$p(style="font-size: 25px;","FDR = NA") 
    },
    
    if (th < 1){
      tags$p(style="color: red",paste0("Warning: With such a dataset size (", nb ," selected discoveries), an FDR of ",round(100*Get_FDR(), digits=2), "% should be cautiously interpreted as strictly less than one discovery (", 
                                       round(th, digits=2), ") 
             is expected to be false")
      )
    } 
  )
  
  
})





output$equivPVal <- renderUI ({
  req(rv$widgets$anaDiff$th_pval)
  
  tags$p(paste0("(p-value = ",signif(10^(- (rv$widgets$anaDiff$th_pval)), digits=3), ")"))
})


output$equivLog10 <- renderText ({
  req(rv$widgets$anaDiff$th_pval)
  
  tags$p(paste0("-log10 (p-value) = ",signif(- log10(rv$widgets$anaDiff$th_pval/100), digits=1)))
})




GetSelectedItems <- reactive({
  req(rv$resAnaDiff)
  rv$widgets$anaDiff$downloadAnaDiff
  
  
  t <- NULL
  upItems1 <- which(-log10(rv$resAnaDiff$P_Value) >=rv$widgets$anaDiff$th_pval)
  upItems2 <- which(abs(rv$resAnaDiff$logFC) >= rv$widgets$hypothesisTest$th_logFC)
  
  if ( rv$widgets$anaDiff$downloadAnaDiff == "All"){
    selectedItems <- 1:nrow(rv$current.obj)
    significant <- rep(0, nrow(rv$current.obj))
    significant[intersect(upItems1, upItems2)] <- 1
  } else {
    selectedItems <- intersect(upItems1, upItems2)
    significant <- rep(1, length(selectedItems))
  }
  
  t <- data.frame(id = rownames(exprs(rv$current.obj))[selectedItems],
                  logFC = round(rv$resAnaDiff$logFC[selectedItems], digits=rv$settings_nDigits),
                  P_Value = rv$resAnaDiff$P_Value[selectedItems],
                  isDifferential = significant)
  tmp <- as.data.frame(fData(rv$current.obj)[selectedItems, rv$widgets$anaDiff$tooltipInfo])
  names(tmp) <- rv$widgets$anaDiff$tooltipInfo
  t <- cbind(t, tmp)
  colnames(t)[2:4] <- paste0(colnames(t)[2:4], " (", as.character(rv$widgets$anaDiff$Comparison),')')
  t
})



isContainedIn <- function(strA, strB){
  return (all(strA %in% strB))
}
samWieczorek/Prostar documentation built on April 27, 2022, 7:32 a.m.