##### SETUP
library(shiny)
library(shinyjs)
library(OmicSelector)
library(dplyr)
library(tools)
library(shinyfullscreen)
suppressMessages(suppressMessages(library(data.table)))
suppressMessages(suppressMessages(library(dplyr)))
suppressMessages(suppressMessages(library(ggplot2)))
suppressMessages(suppressMessages(library(OmicSelector)))
oldwd = getwd()
options(shiny.maxRequestSize = 30*1024^2)
dummy_df = as.data.frame(matrix(rnorm(100),nrow = 10))
library(waiter)
waiting_screen <- tagList(
spin_3(),
h4("OmicSelector is working...")
)
##### INIT VARS
if_analysis_id_duplicated = TRUE
while(if_analysis_id_duplicated == TRUE){
init_random_analysis_id = stringi::stri_rand_strings(1, 15)
if_analysis_id_duplicated = dir.exists(paste0("/OmicSelector/",init_random_analysis_id))
}
##### UI
ui <- fluidPage(
#### STYLE
useShinyjs(),
tags$head(
tags$style(HTML("
#error_msg {
color: red;
}
"))
),
##### MAIN
htmlTemplate("ui.html",
init_random_analysis_id = init_random_analysis_id)
)
##### SERVER
server <- function(input,output,session){
observeEvent(input$button_startover, {
shinyjs::enable("button_start_analysis")
shinyjs::toggle(id= "panelA")
shinyjs::toggle(id= "panelB")
})
observeEvent(input$select_plot, {
output$wykres = renderImage({
filename <- input$select_plot
list(src = filename, width = "100%", height = "100%")
})
})
observeEvent(input$select_df, {
output$download_df = downloadHandler(filename = input$select_df, content = function(file) { file.copy(input$select_df, file) })
})
observeEvent(input$button_start_analysis, {
shinyjs::disable("button_start_analysis")
# VALIDATION:
walidacja = ""
if(dir.exists(paste0("/OmicSelector/",input$analysis_id)) == TRUE) { walidacja = "The analysis with the same analysis ID already exists. Choose a unique one."; updateTextInput(session, "analysis_id", value = stringi::stri_rand_strings(1, 15)); }
if(is.null(input$file2)) { walidacja = "You have to provide xlsx or csv file for the analysis." }
try({
plik = input$file2$datapath
if(tools::file_ext(plik) == "xlsx") { dane = xlsx::read.xlsx(plik,sheetIndex=1) } else {
dane = data.table::fread(plik)
} })
if(is.null(dane)) { walidacja = "You have to provide valid xlsx or csv file for the analysis." }
if(walidacja != "") { showNotification(walidacja, duration = 10, type = "error") }
shinyjs::enable("button_start_analysis")
validate(need(walidacja == "", "Form validation error", "button_start_analysis"))
shinyjs::disable("button_start_analysis")
# Create a Progress object
progress <- shiny::Progress$new()
# Make sure it closes when we exit this reactive, even if there's an error
on.exit(progress$close())
progress$set(message = "Validating submitted data & performing initial analysis", value = 0)
# UNIQUEID:
uniquid = gsub("\\.","",paste0(as.numeric(Sys.time())))
dir.create(paste0(oldwd, "/", uniquid))
setwd(paste0(oldwd, "/", uniquid))
OmicSelector_log(paste0("Staring pre-check of OmicSelector analysis ID: ", getwd()))
# ACTION:
n_steps <- 13
task_status = "OK"
if(file.exists("task.log")) { file.remove("task.log") }
OmicSelector_log("Welcome! OmicSelector id loaded.");
options(warn=-1)
progress$inc(1/n_steps) # PROGRESS BAR 1
writeLines("ERROR", "var_initcheck.txt", sep="")
writeLines(input$class_interest, "var_class_interest.txt", sep="")
writeLines(input$split, "var_split.txt", sep="")
writeLines(input$type, "var_type.txt", sep="")
colnames(dane) = make.names(colnames(dane), unique = T)
dane[dane == ""] = NA
error = FALSE
if("Class" %in% colnames(dane)) { OmicSelector_log("✓ The file contains Class variable. ") } else
{ writeLines(as.character("FAIL"), "var_initcheck.txt", sep=""); OmicSelector_log("☒ The file contains DOES NOT Class variable. "); task_status = "ERROR"; showNotification(type = "error", ui = "☒ The file contains DOES NOT Class variable. ") }
klasy = unique(dane$Class)
ref = c("Case","Control")
if(!dplyr::setequal(ref, klasy)) {
class_interest = readLines("var_class_interest.txt", warn = F)
dane$ClassOrginal = dane$Class
dane$Class = ifelse(dane$Class == class_interest, "Case", "Control")
OmicSelector_log("\n✓ The Class variable was converted successfully. The orginial values are saved in ClassOrginal variable.")
} else {
OmicSelector_log("\n✓ The Class variable has only case and control cases.")
}
progress$inc(1/n_steps)
dane$Class = factor(dane$Class, levels = c("Control","Case"))
if(table(dane$Class)[1] == 0) { writeLines(as.character("FAIL"), "var_initcheck.txt", sep=""); OmicSelector_log("☒ There are no control cases."); task_status = "ERROR"; showNotification(type = "error", ui = "☒ There are no control cases.") }
if(table(dane$Class)[2] == 0) { writeLines(as.character("FAIL"), "var_initcheck.txt", sep=""); OmicSelector_log("☒ There are no cases of interest."); task_status = "ERROR"; showNotification(type = "error", ui = "☒ There are no cases of interest.") }
OmicSelector_log(paste0("\n✓ The data contains ", table(dane$Class)[1], " `Control` cases and ", table(dane$Class)[2], " `Case` cases (cases of interest)."))
progress$inc(1/n_steps)
try({
temp = dplyr::select(dane, starts_with("hsa"))
if(ncol(temp)==0) { writeLines(as.character("FAIL"), "var_initcheck.txt", sep=""); OmicSelector_log("☒ The data does not contain any features (e.g. miRNAs) for feautre selection. Remember that feature names should start from hsa..."); task_status = "ERROR"; showNotification(type = "error", ui = "☒ The data does not contain any features (e.g. miRNAs) for feautre selection. Remember that feature names should start from hsa...") }
OmicSelector_log(paste0("\n✓ The data contains ", ncol(temp), " features (e.g. miRNAs) for selection."))
if(ncol(temp) > 0) { go_next = T } else { go_next = F }
})
progress$inc(1/n_steps)
try({
czy_numeryczne = sapply(temp, is.numeric)
if(sum(czy_numeryczne) != ncol(temp)) { writeLines(as.character("FAIL", "var_initcheck.txt", sep = "")); OmicSelector_log("☒ Some of the features are not numeric. Please remove them. Not numeric: ", paste0(colnames(temp)[czy_numeryczne == F], collapse = ", ")); task_status = "ERROR"; showNotification(type = "error", ui = "☒ Some of the features are not numeric. Please remove them. Not numeric: ", paste0(colnames(temp)[czy_numeryczne == F], collapse = ", "))}
OmicSelector_log(paste0("\n✓ All features are numeric."))
})
progress$inc(1/n_steps)
try({
missing = FALSE
czy_brakna = sapply(temp, is.na)
if(sum(colSums(czy_brakna)) != 0)
{ OmicSelector_log("☒ Some of the features contain missing data.\nWith missing values:\n -", paste0(colnames(temp)[colSums(czy_brakna) > 0], collapse = "\n - ")); task_status = "ERROR"; showNotification(type = "error", ui = "☒ Some of the features contain missing data.\nWith missing values:\n -", paste0(colnames(temp)[colSums(czy_brakna) > 0], collapse = "\n - "))
missing = T } else
{ OmicSelector_log(paste0("\n✓ There are no missing data in features.")) }
})
progress$inc(1/n_steps)
try({
batch = F
if("Batch" %in% colnames(dane)) { OmicSelector_log("\n✓ The file contains `Batch` variable that can be used for batch-effect correction. You can apply combat-based batch correction via OmicSelector, but not via GUI. Please see the package manual. Please also check if the following contingency table is correct:\n ")
print(table(dane$Class, dane$Batch))
batch = T } else { OmicSelector_log("\n✓ The file does not contain `Batch` variable that can be used for batch-effect correction.") }
})
progress$inc(1/n_steps)
try({
x = dplyr::select(dane, starts_with("hsa"))
like_counts = sapply(x, function(x2) (sum(unlist(na.omit(x2))%%1 == 0) + sum(unlist(na.omit(x2)) >= 0))/(2*length(unlist(na.omit(x2)))) == 1)
positive = F
if(sum(like_counts)/ncol(x)) { OmicSelector_log("\n✓ Feature values are positive integers. The file could represent read counts. Please note that the feature selection pipeline requires normalized data. You can use OmicSelector to convert counts to tpm, but not via GUI. Please refer to package manual."); positive = T; } else {
OmicSelector_log("\n✓ Feature values are not positive integers. This is ok if your input data is normalized (e.g. deltaCt values or tpm-normalized counts). Features not looking like counts: ");
OmicSelector_log(paste0(colnames(x)[which(like_counts == FALSE)], collapse = ", "))
}
writeLines(as.character(positive), "var_seemslikecounts.txt", sep="")
})
progress$inc(1/n_steps)
suppressMessages(library(OmicSelector))
try({
split = readLines("var_split.txt", warn = F)
if(split == "yes") {
if("mix" %in% colnames(dane)) {
if(sum(which(dane$mix == 'train')) > 0) {
OmicSelector_log(paste0("\n✓ Samples in training set: ", sum(dane$mix == 'train')));
} else {
OmicSelector_log(paste0("\n✓ Samples in training set: ", sum(which(dane$mix == 'train'))));
OmicSelector_log("There are no samples in training set! Please fix the 'mix' variable. This variable should contain the assignment to 'train', 'test' and 'valid' groups."); task_status = "ERROR"; showNotification(type = "error", ui = "There are no samples in training set! Please fix the 'mix' variable. This variable should contain the assignment to 'train', 'test' and 'valid' groups.")
}
if(sum(which(dane$mix == 'test')) > 0) {
OmicSelector_log(paste0("\n✓ Samples in test set: ", sum(dane$mix == 'test')));
} else {
OmicSelector_log(paste0("\n✓ Samples in test set: ", sum(dane$mix == 'test')));
OmicSelector_log("There are no samples in test set! Please fix the 'mix' variable. This variable should contain the assignment to 'train', 'test' and 'valid' groups."); task_status = "ERROR"; showNotification(type = "error", ui = "There are no samples in test set! Please fix the 'mix' variable. This variable should contain the assignment to 'train', 'test' and 'valid' groups.")
}
if(sum(which(dane$mix == 'valid')) > 0) {
OmicSelector_log(paste0("\n✓ Samples in validation set: ", sum(dane$mix == 'valid')));
} else {
OmicSelector_log(paste0("\n✓ Samples in validation set: ", sum(dane$mix == 'valid')));
OmicSelector_log("There are no samples in validation set! Please fix the 'mix' variable. This variable should contain the assignment to 'train', 'test' and 'valid' groups."); task_status = "ERROR"; showNotification(type = "error", ui = "There are no samples in validation set! Please fix the 'mix' variable. This variable should contain the assignment to 'train', 'test' and 'valid' groups.")
}
train = dplyr::filter(dane, mix == "train")
fwrite(train, "mixed_train.csv")
test = dplyr::filter(dane, mix == "test")
fwrite(test, "mixed_test.csv")
valid = dplyr::filter(dane, mix == "valid")
fwrite(valid, "mixed_valid.csv")
mixed = rbind(train,test,valid)
fwrite(mixed, "data_start.csv")
fwrite(mixed, "mixed.csv")
if(sum(which(dane$mix == 'train_balanced')) > 0) {
train_balanced = dplyr::filter(dane, mix == "train_balanced")
fwrite(train_balanced, "mixed_train_balanced.csv")
OmicSelector_log(paste0("\n✓ Balanced training set file was retored."))
}
dane = OmicSelector_load_datamix(use_smote_not_rose = T) # load mixed_*.csv files
train = dane[[1]]
test = dane[[2]]
valid = dane[[3]]
train_smoted = dane[[4]]
train$mix = "train"
test$mix = "test"
valid$mix = "valid"
train_smoted$mix = "train_balanced"
merged = rbind(train,test,valid, train_smoted)
fwrite(merged, "merged.csv")
} else {
OmicSelector_log("\n✓ The data is not splitted, i.e. doesn't have 'train', 'test' and 'valid' in 'mix' variable. We will perform data splitting (60% train, 20% test, 20% valid). ");
metadane = dplyr::select(dane, -starts_with("hsa"))
ttpm_features = dplyr::select(dane, starts_with("hsa"))
mixed = OmicSelector_prepare_split(metadane = metadane, ttpm = ttpm_features, train_proc = 0.6)
fwrite(mixed, "data_start.csv")
fwrite(mixed, "mixed.csv")
}} else {
fwrite(dane, "data_start.csv")
train = dane
train$mix = "train"
fwrite(train, "mixed_train.csv")
test = dane
test$mix = "test"
fwrite(test, "mixed_test.csv")
valid = dane
valid$mix = "valid"
fwrite(valid, "mixed_valid.csv")
mixed = rbind(train,test,valid)
fwrite(mixed, "mixed.csv")
}
})
progress$inc(1/n_steps)
try({
OmicSelector_log("Checking if data can be loaded. Balancing dataset... (this may take a while)");
dane = OmicSelector_load_datamix(use_smote_not_rose = T) # load mixed_*.csv files
train = dane[[1]]
test = dane[[2]]
valid = dane[[3]]
train_smoted = dane[[4]]
trainx = dane[[5]]
trainx_smoted = dane[[6]] # get the objects from list to make the code more readable.
OmicSelector_log("\n✓ All datasets can be loaded. SMOTE-based balanced dataset can be loaded. ");
})
progress$inc(1/n_steps)
try({
OmicSelector_log("Performing DE analysis...");
type = readLines("var_type.txt", warn = F)
x_mix = dplyr::select(mixed, starts_with("hsa"))
DE_mix = OmicSelector_differential_expression_ttest(x_mix, mixed$Class, mode = type)
fwrite(DE_mix, "DE_mixed.csv")
DE_train = OmicSelector_differential_expression_ttest(trainx, train$Class, mode = type)
fwrite(DE_train, "DE_train.csv")
OmicSelector_log("\n✓ DE was performed for whole dataset (mixed) and for training datasets. ");
})
progress$inc(1/n_steps)
OmicSelector_log("Drawing exploratory plots...");
try({ png("exploratory_pca.png", width = 1170, height = 658)
a = OmicSelector_PCA(trainx, train$Class)
a
ggsave("exploratory_pca.png", a)
suppressMessages(graphics.off()) })
try({ png("exploratory_pca_all_batch.png", width = 1170, height = 658)
a = OmicSelector_PCA(dplyr::select(mixed, starts_with('hsa')), mixed$Batch)
a
ggsave("exploratory_pca_all_batch.png", a)
suppressMessages(graphics.off()) })
try({ png("exploratory_pca_all_mix.png", width = 1170, height = 658)
a = OmicSelector_PCA(dplyr::select(mixed, starts_with('hsa')), mixed$mix)
a
ggsave("exploratory_pca_all_mix.png", a)
suppressMessages(graphics.off()) })
try({ png("exploratory_pca_all_class.png", width = 1170, height = 658)
a = OmicSelector_PCA(dplyr::select(mixed, starts_with('hsa')), mixed$ClassOrginal)
a
ggsave("exploratory_pca_all_class.png", a)
suppressMessages(graphics.off()) })
try({ png("exploratory_vulcano.png", width = 1170, height = 658)
a = OmicSelector_vulcano_plot(DE_train$miR, DE = DE_train, only_label = DE_train$miR[DE_train$`p-value` < 0.05])
a
ggsave("exploratory_vulcano.png", a)
suppressMessages(graphics.off()) })
try({ png("exploratory_vulcano2.png", width = 1170, height = 658)
a = OmicSelector_vulcano_plot(DE_train$miR, DE = DE_train, only_label = DE_train$miR[1:10]) # top 10
a
ggsave("exploratory_vulcano2.png", a)
suppressMessages(graphics.off()) })
try({ png("exploratory_heatmap.png", width = 1170, height = 658)
OmicSelector_heatmap(x = trainx, rlab = data.frame(Class = train$Class), expression_name = "Expression")
suppressMessages(graphics.off()) })
try({ png("exploratory_heatmapsig.png", width = 1170, height = 658)
OmicSelector_heatmap(x = trainx[,DE_train$miR[DE_train$`p-value` < 0.05]], rlab = data.frame(Class = train$Class), expression_name = "Expression")
suppressMessages(graphics.off()) })
try({ png("exploratory_heatmapz.png", width = 1170, height = 658)
OmicSelector_heatmap(x = trainx, rlab = data.frame(Class = train$Class), expression_name = "Expression", zscore = T)
suppressMessages(graphics.off()) })
try({ png("exploratory_heatmapzsig.png", width = 1170, height = 658)
OmicSelector_heatmap(x = trainx[,DE_train$miR[DE_train$`p-value` < 0.05]], rlab = data.frame(Class = train$Class), expression_name = "Expression", zscore = T)
suppressMessages(graphics.off()) })
try({ png("exploratory_heatmapmix.png", width = 1170, height = 658)
OmicSelector_heatmap(x = dplyr::select(mixed, DE_train$miR[DE_train$`p-value` < 0.05]), rlab = data.frame(Class = mixed$Class, Mix = mixed$mix), expression_name = "Expression")
suppressMessages(graphics.off()) })
try({ png("exploratory_heatmapmixz.png", width = 1170, height = 658)
OmicSelector_heatmap(x = dplyr::select(mixed, DE_train$miR[DE_train$`p-value` < 0.05]), rlab = data.frame(Class = mixed$Class, Mix = mixed$mix), expression_name = "Expression", zscore = T)
suppressMessages(graphics.off()) })
if(batch){
try({ png("exploratory_pcab.png", width = 1170, height = 658)
meta = paste0(mixed$Class, " - ", mixed$Batch)
OmicSelector_PCA(dplyr::select(mixed, starts_with("hsa")), meta)
suppressMessages(graphics.off()) })
try({ png("exploratory_heatmapmixb.png", width = 1170, height = 658)
OmicSelector_heatmap(x = dplyr::select(mixed, DE_train$miR[DE_train$`p-value` < 0.05]), rlab = data.frame(Class = mixed$Class, Mix = mixed$mix, Batch = mixed$Batch), expression_name = "Expression")
suppressMessages(graphics.off()) })
try({ png("exploratory_heatmapmixzb.png", width = 1170, height = 658)
OmicSelector_heatmap(x = dplyr::select(mixed, DE_train$miR[DE_train$`p-value` < 0.05]), rlab = data.frame(Class = mixed$Class, Mix = mixed$mix, Batch = mixed$Batch), expression_name = "Expression", zscore = T)
suppressMessages(graphics.off()) })
}
OmicSelector_log("\n✓ PCA, vulcano plot and heatmaps were generated. ");
progress$inc(1/n_steps)
try({
if(!file.exists("data_start.csv")) { fwrite(dane, "data_start.csv") }
# Out: czy missing,czy batch
writeLines(as.character(batch), "var_batch.txt", sep="")
writeLines(as.character(missing), "var_missing.txt", sep="")
writeLines(task_status, "var_initcheck.txt", sep="")
OmicSelector_log("\n✓ The files are ready for further analysis. ");
})
OmicSelector_log("[OmicSelector: TASK COMPLETED]")
output$task_log = shiny::renderPrint(cat(paste0(readLines("temp.log", warn =F), collapse = "\n")))
output$status_log = shiny::renderPrint(cat(paste0(task_status, collapse = "\n")))
wykresy = list.files(".", pattern = "*.png")
updateSelectInput(session, "select_plot", choices = as.list(wykresy))
dfs = list.files(".", pattern = "*.csv")
updateSelectInput(session, "select_df", choices = as.list(dfs))
progress$inc(1/n_steps)
# END:
shinyjs::toggle(id= "panelA")
shinyjs::toggle(id= "panelB")
})
observeEvent(input$button_process, {
if(readLines("var_initcheck.txt", warn = F) != "OK") {
showNotification("Cannot proceed due to file validation errors. Please correct the file and start over.", type = "error")
} else {
showNotification(paste0("Please wait... Creating analysis framework for ID: ", input$analysis_id))
file.rename("temp.log","initial_check.txt")
dir.create(paste0("/OmicSelector/",input$analysis_id))
file.copy(list.files("."), paste0("/OmicSelector/",input$analysis_id), recursive = T)
updateQueryString(paste0("/analysis.php?id=",input$analysis_id), mode = c("replace","push"),
session = getDefaultReactiveDomain())
shinyjs::runjs(paste0('window.location.href = "/analysis.php?id=', input$analysis_id ,'";'))
}
})
}
shinyApp(ui=ui,server=server)
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