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EcoTyper_recovery_visium.R
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EcoTyper_recovery_visium.R
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suppressPackageStartupMessages({
library(config)
library(argparse)
source("pipeline/lib/config.R")
source("pipeline/lib/misc.R")
source("pipeline/lib/multithreading.R")
})
parser <- ArgumentParser(add_help = F)
arguments = parser$add_argument_group('Arguments')
arguments$add_argument("-c", "--config", type = "character", metavar="<PATH>",
help="Path to the config files [required].")
arguments$add_argument("-h", "--help", action='store_true', help="Print help message.")
args <- parser$parse_args()
if(args$h || is.null(args$config))
{
parser$print_help()
quit()
}
config_file = abspath(args$config)
config <- config::get(file = config_file)
discovery = config$Input$"Discovery dataset name"
recovery = config$Input$"Recovery dataset name"
input_path = config$Input$"Input Visium directory"
fractions = config$Input$"Recovery cell type fractions"
coo = config$Input$"Background cell type"
if(is.null(coo))
{
coo = config$Input$"Malignant cell of origin"
}
if(is.null(coo))
{
print(NULL)
coo = "NULL"
}
CSx_username = config$Input$"CIBERSORTx username"
CSx_token = config$Input$"CIBERSORTx token"
n_threads = config$"Pipeline settings"$"Number of threads"
CSx_singularity_path_fractions = config$"Pipeline settings"$"CIBERSORTx fractions Singularity path"
final_output = config$"Output"$"Output folder"
suppressWarnings({
input_path = abspath(input_path)
final_output = abspath(file.path(final_output, recovery))
})
suppressWarnings({
fractions_path = abspath(fractions)
})
if(!is.na(CSx_singularity_path_fractions) && !is.null(CSx_singularity_path_fractions))
{
if(!file.exists(CSx_singularity_path_fractions))
{
stop(paste0("CIBERSORTx fractions Singularity path provided does not exist:", CSx_singularity_path_fractions))
}
}else{
CSx_singularity_path_fractions = "NULL"
}
if(!discovery %in% c("Carcinoma", "Lymphoma"))
{
discovery_config_file = file.path("EcoTyper", discovery, "config_used.yml")
if(!file.exists(discovery_config_file))
{
stop("Error: Cannot read the configuration file used for the discovery of cell states and ecotypes. It should be in the following path: '", config_file, "'. Please make sure that the '--discovery (-d)' argument provided is correct!")
}
config <- config::get(file = discovery_config_file)
discovery_fractions = config$"Pipeline settings"$"Cell type fractions"
if(!is.null(discovery_fractions) && discovery_fractions %in% c("Carcinoma_Fractions", "Lymphoma_Fractions"))
{
fractions = discovery_fractions
}else{
if(!is.null(config$"Pipeline settings"$"Filter genes"))
{
if(config$"Pipeline settings"$"Filter genes" == "cell type specific")
{
fractions = "Cell_type_specific_genes"
}else{
if(config$"Pipeline settings"$"Filter genes" == "no filter")
{
fractions = "All_genes"
}else{
n_genes = as.integer(as.numeric(config$"Pipeline settings"$"Filter genes"))
fractions = paste0("Top_", n_genes)
}
}
}else{
fractions = "Custom"
}
}
}else{
fractions = paste0(discovery, "_Fractions")
}
#Starting EcoTyper
setwd("pipeline")
start = Sys.time()
cat("\nLoading visium data...\n")
PushToJobQueue(paste("Rscript spatial_load_visium_data.R", recovery, paste0("'", input_path, "'")))
RunJobQueue()
if(fractions %in% c("Carcinoma_Fractions", "Lymphoma_Fractions") && !file.exists(fractions_path))
{
cat("\nRunning CIBERSORTxFractions on the visium dataset...\n")
PushToJobQueue(paste("Rscript csx_fractions.R", "visium", recovery, "LM22", "B_mode", CSx_username, CSx_token, paste0("'", CSx_singularity_path_fractions, "'"), TRUE))
RunJobQueue()
PushToJobQueue(paste("Rscript csx_fractions.R", "visium", recovery, "TR4", "B_mode", CSx_username, CSx_token, paste0("'", CSx_singularity_path_fractions, "'"), TRUE))
RunJobQueue()
PushToJobQueue(paste("Rscript csx_fractions_two_tiered.R", "visium", recovery, "TR4", "B_mode", "LM22", "B_mode", fractions))
RunJobQueue()
coo = "Epithelial.cells"
if(fractions %in% "Lymphoma_Fractions")
{
coo = "B.cells"
}
}else{
cat("\nLoading user-provided cell type fractions...\n")
dir.create(file.path("../CIBERSORTx/fractions/visium", recovery, fractions), recursive = T, showWarnings = F)
PushToJobQueue(paste("cp -f '", fractions_path, "' '", file.path("../CIBERSORTx/fractions/visium", recovery, fractions, "CIBERSORTx_Adjusted.txt"), "'"))
RunJobQueue()
}
cat("\nRunning cell state recovery on the visium dataset...\n")
key = read.delim(file.path("../EcoTyper", discovery, fractions, "Analysis", "rank_selection", "rank_data.txt"))
for(cell_type in key[,1])
{
n_states = key[key[,1] == cell_type, 2]
PushToJobQueue((paste("Rscript state_recovery_visium.R", discovery, fractions, cell_type, n_states, recovery, "FALSE")))
}
RunJobQueue()
print(coo)
cat("\nCalculating cell state abundances...\n")
print(paste("Rscript spatial_states.R", discovery, recovery, fractions, coo))
PushToJobQueue((paste("Rscript spatial_states.R", discovery, recovery, fractions, coo)))
RunJobQueue()
cat("\nCalculating ecotype abundances...\n")
PushToJobQueue((paste("Rscript spatial_ecotypes.R", discovery, recovery, fractions, coo)))
RunJobQueue()
cat("\nPlotting cell state heatmaps...\n")
key = read.delim(file.path("../EcoTyper", discovery, fractions, "Analysis", "rank_selection", "rank_data.txt"))
for(cell_type in key[,1])
{
n_states = key[key[,1] == cell_type, 2]
PushToJobQueue((paste("Rscript spatial_plot_states.R", discovery, recovery, fractions, coo, cell_type)))
}
RunJobQueue()
PushToJobQueue((paste("Rscript spatial_plot_ecotypes.R", discovery, recovery, fractions, coo)))
RunJobQueue()
cat("\nCopying EcoTyper results to the output folder!\n")
if(file.exists(final_output) && length(list.files(final_output)) > 0)
{
old_results_folder = paste0(final_output, format(Sys.time(), " %a %b %d %X %Y"))
dir.create(old_results_folder, recursive = T, showWarnings = F)
warning(paste0("The output folder contains files from a previous run. Moving those files to: '", old_results_folder, "'"))
system(paste0("mv -f ", final_output, "/* '", old_results_folder, "'"))
}
dir.create(final_output, recursive = T, showWarnings = F)
key = read.delim(file.path("../EcoTyper", discovery, fractions, "Analysis", "rank_selection", "rank_data.txt"))
for(cell_type in key[,1])
{
system(paste0("cp -f '", file.path("../EcoTyper", discovery, fractions, "Cell_States", "recovery", recovery, paste0(cell_type, "_spatial_heatmaps.pdf")), "' '", final_output, "'"))
system(paste0("cp -f '", file.path("../EcoTyper", discovery, fractions, "Cell_States", "recovery", recovery, paste0(cell_type, "_spatial_heatmaps.png")), "' '", final_output, "'"))
}
system(paste0("cp -f '", file.path("../EcoTyper", discovery, fractions, "Cell_States", "recovery", recovery, "state_abundances.txt"), "' '", final_output, "'"))
system(paste0("cp -f '", file.path("../EcoTyper", discovery, fractions, "Cell_States", "recovery", recovery, "ecotype_abundances.txt"), "' '", final_output, "'"))
system(paste0("cp -f '", file.path("../EcoTyper", discovery, fractions, "Cell_States", "recovery", recovery, "Ecotype_spatial_heatmaps.pdf"), "' '", final_output, "'"))
system(paste0("cp -f '", file.path("../EcoTyper", discovery, fractions, "Cell_States", "recovery", recovery, "Ecotype_spatial_heatmaps.png"), "' '", final_output, "'"))
end = Sys.time()
cat(paste0("\nEcoTyper finished succesfully! Please find the results in: '", final_output, "'.\nRun time: ", format(end - start, digits = 1), "\n"))