# HG changeset patch # User goeckslab # Date 1755113539 0 # Node ID 555ca19d07e6cef933c018480483ba7dae395bf6 planemo upload for repository https://github.com/goeckslab/tools-st/tree/main/tools/spatialge commit 482b2e0e6ca7aaa789ba07b8cd689da9a01532ef diff -r 000000000000 -r 555ca19d07e6 clustering.xml --- /dev/null Thu Jan 01 00:00:00 1970 +0000 +++ b/clustering.xml Wed Aug 13 19:32:19 2025 +0000 @@ -0,0 +1,174 @@ + + Unsupervised spatially-informed clustering on spatialGE spatial transcriptomics data + + macros.xml + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + cluster_plots['plot'] == "cluster_plots" + + + + + + + + + + + + + + + + + + + + + + + + + + + + + diff -r 000000000000 -r 555ca19d07e6 macros.xml --- /dev/null Thu Jan 01 00:00:00 1970 +0000 +++ b/macros.xml Wed Aug 13 19:32:19 2025 +0000 @@ -0,0 +1,16 @@ + + 1.2.2 + 0 + + + + quay.io/goeckslab/spatialge + + + + + + 10.1093/bioinformatics/btac145 + + + \ No newline at end of file diff -r 000000000000 -r 555ca19d07e6 spatialGE_clust.R --- /dev/null Thu Jan 01 00:00:00 1970 +0000 +++ b/spatialGE_clust.R Wed Aug 13 19:32:19 2025 +0000 @@ -0,0 +1,125 @@ +# ------------------------------ +# Spatially-Informed Clustering +# ------------------------------ + +# Purpose: +# Group cells into tissue domains using hierarchical clustering on a weighted similarity matrix + +library(spatialGE) +library(optparse) +library(ggplot2) +library(tools) +library(dplyr) + +### Command Line Options + + +option_list <- list( + make_option(c("-f", "--file"), action = "store", default = NA, type = "character", + help = "STlist .rds file path from preprocessing"), + make_option(c("--visium"), action = "store_true", type = "logical", default = FALSE, + help = "Flag for Visium data"), + make_option(c("--raw"), action = "store_true", type = "logical", default = FALSE, + help = "Flag for raw data"), + make_option(c("--cosmx"), action = "store_true", type = "logical", default = FALSE, + help = "Flag for CosMX data"), + make_option(c("-s", "--samples"), action = "store", default = NULL, type = "character", + help = "Sample subset to perform clustering on"), + make_option(c("-w", "--weight"), action = "store", default = 0.025, type = "numeric", + help = "Weight to be applied to spatial distances between 0-1"), + make_option(c("-d", "--dist"), action = "store", default = "euclidean", type = "character", + help = "Distance metric to be used, all methods found in wordspace::dist.matrix"), + make_option(c("--dslogical"), action = "store_true", type = "logical", default = FALSE, + help = "Deepsplit logical flag"), + make_option(c("--logical"), action = "store", default = TRUE, type = "logical", + help = "Control cluster resolution, true will produce more clusters"), + make_option(c("--dsnumeric"), action = "store_true", type = "logical", default = FALSE, + help = "Deepsplit numeric flag"), + make_option(c("--numeric"), action = "store", default = 0, type = "numeric", + help = "control cluster resolution between 0-4, higher number will produce more clusters"), + make_option(c("-p", "--ptsize"), action = "store", default = 2.75, type = "numeric", + help = "Size of points on cluster plot") +) + +### Main + +# parse args +opt <- parse_args(OptionParser(option_list = option_list)) + +# read in ST data from spatialGE preprocessing +STdata <- readRDS(opt$file) + +message("Rds object successfully loaded") + +# if deepSplit flag is included, choose between logical or numeric +if (opt$logical) { + deepsplit <- opt$dslogical +} else if (opt$numeric) { + deepsplit <- opt$dsnumeric +} + +if (!is.null(opt$samples)) { + opt$samples <- strsplit(opt$samples, ",")[[1]] +} else if (is.null(opt$samples)) { + opt$samples <- NULL +} + +# perform data clustering on transformed data +clusters <- STclust(x = STdata, samples = opt$samples, ws = opt$weight, dist_metric = opt$dist, deepSplit = deepsplit) + +message("Unsupervised spatially-informed clustering has been performed") + +# transform S4 to list for easier slot access +S4toList <- function(obj) { + slot_names <- slotNames(obj) + structure(lapply(slot_names, slot, object = obj), names = slot_names) +} + +STdata <- S4toList(STdata) + +message("Transformed to S4 for slot name access") + +# depending on data input type, pull correct sample column +if (opt$visium) { + sample_col <- "sample_id" +} else if (opt$raw) { + sample_col <- "sampleID" +} else if (opt$cosmx) { + sample_col <- "sample_name" +} + + +if (!is.null(opt$samples)) { + samples <- STdata$sample_meta %>% pull(.data[[sample_col]]) %>% intersect(opt$samples) +} else { + samples <- STdata$sample_meta %>% pull(.data[[sample_col]]) +} + + +message("Sample names identified") + +# create cluster plot directory +if (!dir.exists("cluster_plots")) dir.create("cluster_plots") + +# iterate through each sample, generate a plot, and save +for (s in samples) { + + plot <- STplot(x = clusters, ws = opt$weight, ptsize = opt$ptsize, deepSplit = deepsplit, samples = s) + + message("Cluster plots generated") + + + #create unique plot file names based on sample name + + filename <- paste0("clustered_", s, ".png") + + #save plot to subdir + ggsave( + path = "./cluster_plots", + filename = filename, + bg = "white", + width = 12 + ) +} + +message("Cluster plots saved") diff -r 000000000000 -r 555ca19d07e6 spatialGE_multiple_input.R --- /dev/null Thu Jan 01 00:00:00 1970 +0000 +++ b/spatialGE_multiple_input.R Wed Aug 13 19:32:19 2025 +0000 @@ -0,0 +1,197 @@ +# ------------- +# Data Cleaning +# ------------- + +# MULTIPLE INPUT SCRIPT: +# Accepts multiple sample input for raw data and cosmx +# Accepts single and multiple sample input for Visium, due to spatial subdirectory + +# Purpose: +# Transform data into STlist, perform QC, log transform + +library(spatialGE) +library(optparse) +library(ggplot2) +library(tools) +library(fs) + + +### Command Line Options + +option_list <- list( + make_option(c("-c", "--counts"), action = "store", default = NA, type = "character", + help = "Path to count data file(s)"), + make_option(c("-s", "--spots"), action = "store", default = NULL, type = "character", + help = "Path to cell coordinates file(s), not required for Visium or Xenium"), + make_option(c("-m", "--meta"), action = "store", default = NA, type = "character", + help = "Path to metadata file"), + make_option(c("-n", "--names"), action = "store", default = NA, type = "character", + help = "Specific sample names"), + make_option(c("--plotmeta"), action = "store", default = NULL, type = "character", + help = "Plots counts per cell or genes per cell"), + make_option(c("--samples"), action = "store", default = NULL, type = "character", + help = "Samples to include in plots, defaults to all"), + make_option(c("--sminreads"), action = "store", default = 0, type = "integer", + help = "Minimum number of total reads for a spot to be retained"), + make_option(c("--smaxreads"), action = "store", default = NULL, type = "integer", + help = "Maximum number of total reads for a spot to be retained"), + make_option(c("--smingenes"), action = "store", default = 0, type = "integer", + help = "Minimum number of non-zero counts for a spot to be retained"), + make_option(c("--smaxgenes"), action = "store", default = NULL, type = "integer", + help = "Maximum number of non-zero counts for a spot to be retained"), + make_option(c("--gminreads"), action = "store", default = 0, type = "integer", + help = "Minimum number of total reads for a gene to be retained"), + make_option(c("--gmaxreads"), action = "store", default = NULL, type = "integer", + help = "Maximum number of total reads for a gene to be retained"), + make_option(c("--gminspots"), action = "store", default = 0, type = "integer", + help = "Minimum number of spots with non-zero counts for a gene to be retained"), + make_option(c("--gmaxspots"), action = "store", default = NULL, type = "integer", + help = "Maximum number of spots with non-zero counts for a gene to be retained"), + make_option(c("--distplot"), action = "store_true", type = "logical", default = FALSE, + help = "If set, generate unfiltered distribution plot"), + make_option(c("--filter"), action = "store_true", type = "logical", default = FALSE, + help = "If set, apply filtering before transformation"), + make_option(c("--filterplot"), action = "store_true", type = "logical", default = FALSE, + help = "If set, generate filtered distribution plot"), + make_option(c("-t", "--type"), action = "store_true", default = "log", type = "character", + help = "Type of transformation to apply: log or sct") +) + +### Main + +#parse args +opt <- parse_args(OptionParser(option_list = option_list)) + +#check if metadata or sample names were provided +#need metadata for raw and visium data, sample names for cosmx +if (!is.na(opt$meta) && is.na(opt$names)) { + samples_input <- opt$meta +} else if (is.na(opt$meta) && !is.na(opt$names)) { + samples_input <- unlist(strsplit(opt$names, split = ",")) +} else { + stop("Please only specify either --metadata OR --names") +} + +#create temporary directory to hold count data +count_dir <- tempdir() +unlink(count_dir, recursive = TRUE) +dir.create(count_dir) + +#create temporary directory to hold coord data +coord_dir <- tempdir() +unlink(coord_dir, recursive = TRUE) +dir.create(coord_dir) + +#if spotcoords were provided, load in count and coord data +#if spotcoords were not provided (visium input), only load the count file +if (!is.null(opt$spots)) { + coord_dir <- as.character(opt$spots) + coord_file <- fs::dir_ls(coord_dir) + count_dir <- as.character(opt$counts) + count_file <- fs::dir_ls(count_dir) +} else { + count_dir <- as.character(opt$counts) + count_file <- fs::dir_ls(count_dir) +} + +#if spotcoords are present, include in stlist input +if (!is.null(opt$spots)) { + st_data <- STlist(rnacounts = count_file, spotcoords = coord_file, samples = samples_input) +} else { + st_data <- STlist(rnacounts = count_file, samples = samples_input) +} + +message("STList has been created") + +#distribution plot + +#create distribution plot if flag is included +if (opt$distplot) { + + #if sample names are provided, separate the character string + if (!is.null(opt$samples) && opt$samples != "") { + sample_names <- strsplit(opt$samples, split = ",", fixed = TRUE)[[1]] + } else { + sample_names <- NULL + } + + #generate distribution plot + dist_plot <- distribution_plots(x = st_data, plot_meta = opt$plotmeta, samples = sample_names, ptsize = 1) + + #create unique plot file names based on sample name + base_input <- basename(opt$counts) + base_name <- file_path_sans_ext(base_input) + + filename <- paste0("unfiltered_", base_name, ".png") + + #create output directory for distribution plots + dir.create("./unfiltered_distribution_plots", showWarnings = FALSE, recursive = TRUE) + + #save plot to subdir + ggsave( + path = "./unfiltered_distribution_plots", + filename = filename, + bg = "white", + width = 12 + ) + + message("Unfiltered distribution plot saved to ./unfiltered_distribution_plots") +} + +#spot/cell filtering + +#filter spots if flag is included +if (opt$filter) { + + #filter out spots or genes based on minimum and maximum counts + st_data <- filter_data(x = st_data, spot_minreads = opt$sminreads, spot_maxreads = opt$smaxreads, spot_mingenes = opt$smingenes, + spot_maxgenes = opt$smaxgenes, gene_minreads = opt$gminreads) + message("Data filtering completed & saved to STlist") +} + +#filtered data plot + +#create filtered distribution plot if flag is included +if (opt$filterplot) { + + #if sample names are provided, separate the character string + if (!is.null(opt$samples) && opt$samples != "") { + sample_names <- strsplit(opt$samples, split = ",", fixed = TRUE)[[1]] + } else { + sample_names <- NULL + } + + #generate filtered distribution plot + filter_dist_plot <- distribution_plots(x = st_data, plot_meta = opt$plotmeta, samples = sample_names, ptsize = 1) + + #create unique plot file names based on sample name + base_input_2 <- basename(opt$counts) + base_name_2 <- file_path_sans_ext(base_input_2) + + filename_2 <- paste0("filtered_", base_name_2, ".png") + + #create output directory for cluster plots + dir.create("./filtered_distribution_plots", showWarnings = FALSE, recursive = TRUE) + + #save plot to subdir + ggsave( + path = "./filtered_distribution_plots", + filename = filename_2, + bg = "white", + width = 12 + ) + + message("Filtered distribution plot saved to ./filtered_distribution_plots") +} + +#transform data, defaults to log transformation + +STobj <- transform_data(x = st_data, method = opt$type) + +message("Data has been log transformed, unless otherwise specified") + +#save transformed data to .rds + +saveRDS(STobj, file = "STobj.rds") + +message("STlist has been saved as .rds file") diff -r 000000000000 -r 555ca19d07e6 spatialGE_single_input.R --- /dev/null Thu Jan 01 00:00:00 1970 +0000 +++ b/spatialGE_single_input.R Wed Aug 13 19:32:19 2025 +0000 @@ -0,0 +1,174 @@ +# ------------- +# Data Cleaning +# ------------- + +# SINGLE INPUT SCRIPT: +# Accepts single raw data sample and single cosmx sample +# Does not accept single visium sample due to spatial subdirectory + +# Purpose: +# Transform data into STlist, perform QC, log transform + +library(spatialGE) +library(optparse) +library(ggplot2) +library(tools) +library(fs) + + +### Command line options + +option_list <- list( + make_option(c("-c", "--counts"), action = "store", default = NA, type = "character", + help = "Path to count data file(s)"), + make_option(c("-s", "--spots"), action = "store", default = NULL, type = "character", + help = "Path to cell coordinates file(s), not required for Visium or Xenium"), + make_option(c("-m", "--meta"), action = "store", default = NA, type = "character", + help = "Path to metadata file"), + make_option(c("-n", "--names"), action = "store", default = NA, type = "character", + help = "Specific sample names"), + make_option(c("--plotmeta"), action = "store", default = NULL, type = "character", + help = "Plots counts per cell or genes per cell"), + make_option(c("--samples"), action = "store", default = NULL, type = "character", + help = "Samples to include in plots, defaults to all"), + make_option(c("--sminreads"), action = "store", default = 0, type = "integer", + help = "Minimum number of total reads for a spot to be retained"), + make_option(c("--smaxreads"), action = "store", default = NULL, type = "integer", + help = "Maximum number of total reads for a spot to be retained"), + make_option(c("--smingenes"), action = "store", default = 0, type = "integer", + help = "Minimum number of non-zero counts for a spot to be retained"), + make_option(c("--smaxgenes"), action = "store", default = NULL, type = "integer", + help = "Maximum number of non-zero counts for a spot to be retained"), + make_option(c("--gminreads"), action = "store", default = 0, type = "integer", + help = "Minimum number of total reads for a gene to be retained"), + make_option(c("--gmaxreads"), action = "store", default = NULL, type = "integer", + help = "Maximum number of total reads for a gene to be retained"), + make_option(c("--gminspots"), action = "store", default = 0, type = "integer", + help = "Minimum number of spots with non-zero counts for a gene to be retained"), + make_option(c("--gmaxspots"), action = "store", default = NULL, type = "integer", + help = "Maximum number of spots with non-zero counts for a gene to be retained"), + make_option(c("--distplot"), action = "store_true", type = "logical", default = FALSE, + help = "If set, generate unfiltered distribution plot"), + make_option(c("--filter"), action = "store_true", type = "logical", default = FALSE, + help = "If set, apply filtering before transformation"), + make_option(c("--filterplot"), action = "store_true", type = "logical", default = FALSE, + help = "If set, generate filtered distribution plot"), + make_option(c("-t", "--type"), action = "store_true", default = "log", type = "character", + help = "Type of transformation to apply: log or sct") +) + +### Main + +#parse args +opt <- parse_args(OptionParser(option_list = option_list)) + +#check if metadata or sample names were provided +#need metadata for raw, sample names for cosmx +if (!is.na(opt$meta) && is.na(opt$names)) { + samples_input <- opt$meta +} else if (is.na(opt$meta) && !is.na(opt$names)) { + samples_input <- opt$names +} else { + stop("Please only specify either --metadata OR --names") +} + +#create STlist with single input flags +st_data <- STlist(rnacounts = opt$counts, spotcoords = opt$spots, samples = samples_input) + +message("STlist has been created") + +#distribution plot + +#create distribution plot if flag is included +if (opt$distplot) { + + #if sample names are provided, separate the character string + #probably don't need strsplit, keeping for safety + if (!is.null(opt$samples) && opt$samples != "") { + sample_names <- strsplit(opt$samples, split = ",", fixed = TRUE)[[1]] + } else { + sample_names <- NULL + } + + #generate distribution plot + dist_plot <- distribution_plots(x = st_data, plot_meta = opt$plotmeta, samples = sample_names, ptsize = 1) + + #create unique plot file names based on sample name + base_input <- basename(opt$counts) + base_name <- file_path_sans_ext(base_input) + + filename <- paste0("unfiltered_", base_name, ".png") + + #create output directory for cluster plots + dir.create("./unfiltered_distribution_plots", showWarnings = FALSE, recursive = TRUE) + + #save plot to subdir + ggsave( + path = "./unfiltered_distribution_plots", + filename = filename, + bg = "white", + width = 12 + ) + + message("Unfiltered distribution plot saved to ./unfiltered_distribution_plots") +} + +#spot/cell filtering + +#filter spots if flag is included +if (opt$filter) { + + #filter out spots or genes based on minimum and maximum counts + st_data <- filter_data(x = st_data, spot_minreads = opt$sminreads, spot_maxreads = opt$smaxreads, spot_mingenes = opt$smingenes, + spot_maxgenes = opt$smaxgenes, gene_minreads = opt$gminreads) + + message("Data filtering completed & saved to STlist") +} + +#filtered data plot + +#create filtered distribution plot if flag is included +if (opt$filterplot) { + + #if sample names are provided, separate the character string + #probably don't need strsplit, keeping for safety + if (!is.null(opt$samples) && opt$samples != "") { + sample_names <- strsplit(opt$samples, split = ",", fixed = TRUE)[[1]] + } else { + sample_names <- NULL + } + + #generate filtered distribution plot + filter_dist_plot <- distribution_plots(x = st_data, plot_meta = opt$plotmeta, samples = sample_names, ptsize = 1) + + #create unique plot file names based on sample name + base_input_2 <- basename(opt$counts) + base_name_2 <- file_path_sans_ext(base_input_2) + + filename_2 <- paste0("filtered_", base_name_2, ".png") + + #create output directory for cluster plots + dir.create("./filtered_distribution_plots", showWarnings = FALSE, recursive = TRUE) + + #save plot to subdir + ggsave( + path = "./filtered_distribution_plots", + filename = filename_2, + bg = "white", + width = 12 + ) + + message("Filtered distribution plot saved to ./filtered_distribution_plots") +} + +#transform data, defaults to log transformation + +STobj <- transform_data(x = st_data, method = opt$type) + +message("Data has been log transformed, unless otherwise specified") + +#save transformed data to .rds + +saveRDS(STobj, file = "STobj.rds") + +message("STlist has been saved as .rds file") diff -r 000000000000 -r 555ca19d07e6 test-data/ST_mel3_rep1_counts.tsv --- /dev/null Thu Jan 01 00:00:00 1970 +0000 +++ b/test-data/ST_mel3_rep1_counts.tsv Wed Aug 13 19:32:19 2025 +0000 @@ -0,0 +1,1000 @@ +gene 5x8 6x8 6x9 6x10 7x7 7x8 7x9 7x10 7x11 7x12 8x7 8x8 8x9 8x10 8x11 8x12 8x13 8x14 9x6 9x7 9x8 9x9 9x10 9x11 9x12 9x13 9x14 9x15 9x16 9x17 10x6 10x7 10x8 10x9 10x10 10x11 10x12 10x13 10x15 10x16 10x17 10x18 10x19 11x6 11x7 11x8 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