biopipen 0.33.0__py3-none-any.whl → 0.34.0__py3-none-any.whl

biopipen/scripts/scrna/ExprImputation-alra.R

@@ -1,30 +1,106 @@
-{{ biopipen_dir | joinpaths: "utils", "misc.R" | source_r }}
-
 library(SeuratWrappers)
 library(Seurat)
+library(purrr)
+library(stringr)
+library(biopipen.utils)
 
 infile = {{in.infile | r}}
 outfile = {{out.outfile | r}}
 envs = {{envs.alra_args | r}}
 
-log_info("Loading Seurat object")
-sobj = readRDS(infile)
+log <- get_logger()
+
+log$info("Loading Seurat object")
+sobj <- read_obj(infile)
 assay <- DefaultAssay(sobj)
 
-log_info("Imputing expression values, using ALRA")
-envs$object = sobj
+# https://github.com/mojaveazure/seurat-disk/issues/102
+# https://github.com/simoncmo/shared_seurat_scripts/blob/main/function_seurat_janitor.R
+# Try to fix the issue with SCTModel
+log$info("Trying to fix SCTModel issue (see mojaveazure/seurat-disk#102)")
+# --------------------------------------------------------------------------
+# Handle missing median_umi
+fix_median_umi = function(SCTModel_obj){
+	err_message = ''
+	tryCatch({ test <- methods::validObject(SCTModel_obj) },
+		error = function(error_message) {
+			err_message <<- as.character(error_message)
+	})
+	missing_medium_umi = stringr::str_detect(err_message, 'median_umi')
+
+	if(missing_medium_umi){
+		message('Missing medium_umi, calculate again from cell.attributes$umi')
+		slot(SCTModel_obj, 'median_umi') = median(SCTModel_obj@cell.attributes$umi)
+	}
+	return(SCTModel_obj)
+}
+
+# Cleaning empty objects
+# General purpose
+clean_seurat_obj_list = function(obj_list, attirbute_to_check){
+	if(missing(attirbute_to_check)) {stop("Need attributes to check for cleaning")}
+	# Object type
+	obj_type = class(obj_list[[1]])[[1]]
+
+	# Count
+	obj_size = unlist(purrr::map(obj_list, function(object){
+		nrow(slot(object, attirbute_to_check))
+	}))
+
+	# Remove empty
+	if(length(obj_size ==0)  != 0 ){
+		message(str_glue('Removing {length(obj_size ==0)} empty object from the {obj_type} object list'))
+		obj_list = obj_list[obj_size!=0]
+		message(str_glue('{length(obj_list)} {obj_type} object(s) left'))
+	}
+	obj_list
+}
+
+# for SCTModel.list slot
+clean_seurat_SCTModel_list = function(sct_model_list){
+	clean_seurat_obj_list(obj_list = sct_model_list, attirbute_to_check = 'cell.attributes')
+}
+
+fix_seurat_SCT = function(obj){
+	# Check first
+	if(!'SCT' %in% Assays(obj)){
+		message('SCT assay not found. Nothing to fix')
+		return(obj)
+	}
+
+	# Model list
+	sct_model_list = obj$SCT@SCTModel.list
+	# 1. clean SCTModel list
+	sct_model_list = clean_seurat_SCTModel_list(sct_model_list)
+
+	# 2. fix missing median_umi
+	sct_model_list = map(sct_model_list, function(sct_model){
+		fix_median_umi(sct_model)
+	})
+
+	# Add back and retrun
+	obj$SCT@SCTModel.list = sct_model_list
+
+	return(obj)
+}
+# --------------------------------------------------------------------------
+sobj = fix_seurat_SCT(sobj)
+
+log$info("Imputing expression values, using ALRA")
+envs$object <- sobj
 sobj = do_call(RunALRA, envs)
+envs$object <- NULL
+gc()
 
-# sobj = RunALRA(sobj)
-log_info("Renaming assays")
+log$info("Renaming assays")
 sobj = RenameAssays(sobj, assay.name = assay, new.assay.name = "RAW")
 sobj = RenameAssays(sobj, assay.name = "alra", new.assay.name = assay)
 DefaultAssay(sobj) <- assay
 
-sobj@misc$impute = "alra"
+sobj@misc$impute_method = "alra"
 
-log_info("Saving Seurat object")
-saveRDS(sobj, outfile)
+log$info("Saving Seurat object")
+save_obj(sobj, outfile)
 
 # choosek_plot_file = file.path(dirname(outfile), "choosek.png")
 # png(choosek_plot_file, width = 1200, height = 1000, res = 100)

biopipen/scripts/scrna/ExprImputation-rmagic.R

@@ -1,30 +1,256 @@
+tryCatch(
+    {
+        # in order to load Rmagic
+        workdir <- {{ job.outdir | r }}
+        conda_prefix <- Sys.getenv("CONDA_PREFIX")
+        setwd(workdir)
+        if (!dir.exists("miniconda3")) {
+            file.symlink(conda_prefix, "miniconda3")
+        }
+    },
+    error = function(e) {}
+)
 
-tryCatch({
-    # in order to load Rmagic
-    workdir = {{job.outdir | r}}
-    conda_prefix = Sys.getenv("CONDA_PREFIX")
-    setwd(workdir)
-    file.symlink(conda_prefix, "miniconda3")
-}, error=function(e) {})
-
-python = {{envs.rmagic_args.python | r}}
-Sys.setenv(RETICULATE_PYTHON = Sys.which(python))
-# reticulate::use_python(python)
+python <- {{ envs.rmagic_args.python | r }}
+Sys.setenv(RETICULATE_PYTHON = ifelse(grepl("/", python, fixed = TRUE), python, Sys.which(python)))
+# reticulate::use_python(python, require = TRUE)
 
 library(Rmagic)
+Rmagic:::load_pymagic()
+pymagic <- tryCatch({
+    Rmagic:::pymagic
+}, error = function(e) {
+    NULL
+})
+if (is.null(pymagic)) {
+    stop("Failed to load pymagic module. Please check your Python environment.\n ",
+         "Current python used by reticulate: ", reticulate::py_config()$python)
+}
+
+library(Matrix)
 library(Seurat)
+library(biopipen.utils)
+
+log <- get_logger()
+
+infile <- {{ in.infile | r }}
+outfile <- {{ out.outfile | r }}
+threshold <- {{ envs.rmagic_args.threshold | r }}
+
+log$info("Loading Seurat object ...")
+sobj <- read_obj(infile)
+
+if (threshold > 0) {
+    # only use the genes with expression in number of cells greater than threshold
+    log$info("Fetching genes with expression great than threshold ({threshold}) ...")
+    # get the expression matrix
+    layers <- Layers(sobj)
+    layer <- ifelse(!"counts" %in% layers, "data", "counts")
+    counts <- GetAssayData(sobj, layer = layer)
+    # Percent of cells expressing each gene
+    dropout_rates <- Matrix::rowSums(counts == 0) / ncol(counts)
+
+    # Genes to impute
+    genes_to_impute <- names(dropout_rates[dropout_rates > threshold])
+
+    log$info("- Will impute for {length(genes_to_impute)}/{length(dropout_rates)} genes ...")
+    rm(counts)
+    rm(dropout_rates)
+    gc()
+} else {
+    genes_to_impute <- NULL
+}
+
+# get the expression matrix
+data_impute <- t(GetAssayData(sobj, layer = "data"))
+
+log$info("Running MAGIC ...")
+check.int.or.null <- function(x) {
+    if (is.numeric(x = x)) {
+        x <- as.integer(x = x)
+    } else if (!is.null(x = x) && is.na(x = x)) {
+        x <- NULL
+    }
+    x
+}
+
+check.double.or.null <- function(x) {
+    if (is.numeric(x = x)) {
+        x <- as.integer(x = x)
+    } else if (!is.null(x = x) && is.na(x = x)) {
+        x <- NULL
+    }
+    x
+}
+
+check.int.or.string <- function(x, str) {
+    if (is.numeric(x = x)) {
+        x <- as.integer(x = x)
+    } else if (is.null(x = x) || is.na(x = x)) {
+        x <- str
+    }
+    x
+}
+# the magic function is defined in the Rmagic package
+# it has a bug at line 138 when genes are given as a character vector
+# See also https://github.com/KrishnaswamyLab/MAGIC/issues/227
+magic_patched <- function(
+    data,
+    genes = NULL,
+    knn = 5,
+    knn.max = NULL,
+    decay = 1,
+    t = 3,
+    npca = 100,
+    solver = "exact",
+    init = NULL,
+    t.max = 20,
+    knn.dist.method = "euclidean",
+    verbose = 1,
+    n.jobs = 1,
+    seed = NULL,
+    # deprecated args
+    k = NULL, alpha = NULL,
+    ...) {
+    # check installation
+    # if (!reticulate::py_module_available(module = "magic") ||
+    #     !exists("pymagic") || is.null(pymagic)) {
+    #     Rmagic:::load_pymagic()
+    # }
+    # check for deprecated arguments
+    if (!is.null(k)) {
+        message("Argument k is deprecated. Using knn instead.")
+        knn <- k
+    }
+    if (!is.null(alpha)) {
+        message("Argument alpha is deprecated. Using decay instead.")
+        decay <- alpha
+    }
+    # validate parameters
+    knn <- as.integer(x = knn)
+    t.max <- as.integer(x = t.max)
+    n.jobs <- as.integer(x = n.jobs)
+    npca <- check.int.or.null(npca)
+    knn.max <- check.int.or.null(knn.max)
+    seed <- check.int.or.null(seed)
+    verbose <- check.int.or.null(verbose)
+    decay <- check.double.or.null(decay)
+    t <- check.int.or.string(t, "auto")
+    if (!methods::is(object = data, "Matrix")) {
+        data <- as.matrix(x = data)
+    }
+    # if (length(genes) <= 1 && (is.null(x = genes) || is.na(x = genes))) {
+    #                                                  ^^^^^^^^^^^^^^^^ bug here
+    if (length(genes) <= 1 && (is.null(x = genes) || (length(genes) == 1 && is.na(x = genes)))) {
+        genes <- NULL
+        gene_names <- colnames(x = data)
+    } else if (is.numeric(x = genes)) {
+        gene_names <- colnames(x = data)[genes]
+        genes <- as.integer(x = genes - 1)
+    } else if (length(x = genes) == 1 && genes == "all_genes") {
+        gene_names <- colnames(x = data)
+    } else if (length(x = genes) == 1 && genes == "pca_only") {
+        gene_names <- paste0("PC", 1:npca)
+    } else {
+        # character vector
+        if (!all(genes %in% colnames(x = data))) {
+            warning(paste0(
+                "Genes ",
+                genes[!(genes %in% colnames(data))],
+                " not found.",
+                collapse = ", "
+            ))
+        }
+        genes <- which(x = colnames(x = data) %in% genes)
+        gene_names <- colnames(x = data)[genes]
+        genes <- as.integer(x = genes - 1)
+    }
+    # store parameters
+    params <- list(
+        "data" = data,
+        "knn" = knn,
+        "knn.max" = knn.max,
+        "decay" = decay,
+        "t" = t,
+        "npca" = npca,
+        "solver" = solver,
+        "knn.dist.method" = knn.dist.method
+    )
+    # use pre-initialized values if given
+    operator <- NULL
+    if (!is.null(x = init)) {
+        if (!methods::is(init, "magic")) {
+            warning("object passed to init is not a phate object")
+        } else {
+            operator <- init$operator
+            operator$set_params(
+                knn = knn,
+                knn_max = knn.max,
+                decay = decay,
+                t = t,
+                n_pca = npca,
+                solver = solver,
+                knn_dist = knn.dist.method,
+                n_jobs = n.jobs,
+                random_state = seed,
+                verbose = verbose,
+                ...
+            )
+        }
+    }
+    if (is.null(x = operator)) {
+        operator <- pymagic$MAGIC(
+            knn = knn,
+            knn_max = knn.max,
+            decay = decay,
+            t = t,
+            n_pca = npca,
+            solver = solver,
+            knn_dist = knn.dist.method,
+            n_jobs = n.jobs,
+            random_state = seed,
+            verbose = verbose,
+            ...
+        )
+    }
+    result <- operator$fit_transform(
+        data,
+        genes = genes,
+        t_max = t.max
+    )
+    colnames(x = result) <- gene_names
+    rownames(x = result) <- rownames(data)
+    result <- as.data.frame(x = result)
+    result <- list(
+        "result" = result,
+        "operator" = operator,
+        "params" = params
+    )
+    class(x = result) <- c("magic", "list")
+    return(result)
+}
 
-infile <- {{in.infile | r}}
-outfile <- {{out.outfile | r}}
+data_impute <- magic_patched(data_impute, genes = genes_to_impute)
 
-sobj <- readRDS(infile)
-assay <- DefaultAssay(sobj)
+if (threshold > 0) {
+    data <- t(GetAssayData(sobj, layer = "data"))
+    data_impute <- cbind(data[, setdiff(colnames(data), genes_to_impute)], Matrix::as.matrix(data_impute$result))
+    rm(data)
+    gc()
+} else {
+    # if threshold is 0, then we need to transpose the data back
+    data_impute <- t(Matrix::as.matrix(data_impute$result))
+}
 
-sobj <- magic(sobj)
-sobj <- RenameAssays(sobj, assay.name = assay, new.assay.name = "RAW")
-sobj <- RenameAssays(sobj, assay.name = "MAGIC_RNA", new.assay.name = assay)
+log$info("Adding imputed data to Seurat object ...")
+# Add imputed data to the Seurat object
+sobj <- SetAssayData(
+    sobj,
+    layer = "data",
+    new.data = t(data_impute)
+)
 
-DefaultAssay(sobj) <- assay
+sobj@misc$impute_method <- "rmagic"
 
-attr(sobj, "impute") = "rmagic"
-saveRDS(sobj, outfile)
+log$info("Saving Seurat object ...")
+save_obj(sobj, outfile)

biopipen/scripts/scrna/ExprImputation-scimpute.R

@@ -3,7 +3,7 @@ library(Seurat)
 
 infile = {{in.infile | r}}
 outfile = {{out.outfile | r}}
-joboutdir = "{{job.outdir}}/"
+joboutdir = {{job.outdir | append: "/" | r}}
 drop_thre = {{envs.scimpute_args.drop_thre | r}}
 kcluster = {{(envs.scimpute_args.kcluster | default: None | r}}
 ncores = {{envs.scimpute_args.ncores | r}}
@@ -12,7 +12,7 @@ refgene = {{envs.scimpute_args.refgene | r}}
 setwd(joboutdir)
 
 labels = NULL
-sobj = readRDS(infile)
+sobj = read_obj(infile)
 counts = as.data.frame(sobj@assays$RNA@counts)
 kc = length(unique(Idents(sobj)))
 if (kc > 0) {
@@ -38,6 +38,9 @@ scimpute(
 imputed = readRDS(file.path(joboutdir, "scimpute_count.rds"))
 outobj = CreateSeuratObject(counts = imputed)
 
-outobj@meta.data = sobj@meta.data[rownames(outobj@meta.data),,drop=FALSE]
-attr(outobj, "impute") = "scimpute"
-saveRDS(outobj, outfile)
+outobj@meta.data = sobj@meta.data[rownames(outobj@meta.data), , drop=FALSE]
+# remember that it is the counts being imputed, we still need to
+# normalize the data
+outobj@misc$impute_method = "scimpute"
+
+save_obj(outobj, outfile)

biopipen/scripts/scrna/LoomTo10X.R

@@ -0,0 +1,51 @@
+library(loomR)
+library(DropletUtils)
+library(Matrix)
+
+loomfile <- {{in.loomfile | r}}
+outdir <- {{out.outdir | r}}
+
+lfile <- connect(filename = loomfile, mode = "r")
+
+# Extract the expression matrix (genes x cells)
+expr_matrix <- t(lfile[["matrix"]][, ])
+if (!inherits(expr_matrix, "dgCMatrix")) {
+    expr_matrix <- Matrix::Matrix(expr_matrix, sparse = TRUE)
+}
+
+# Extract gene names and IDs
+gene_names <- lfile[["row_attrs/Gene"]][]
+
+gene_ids <- tryCatch({
+   lfile[["row_attrs/GeneID"]][]
+}, error = function(e) {
+   NULL
+})
+
+if (is.null(gene_ids)) {
+  gene_ids <- gene_names
+}
+
+# Extract cell barcodes
+cell_barcodes <- lfile[["col_attrs/CellID"]][]
+
+# Close the LOOM file connection
+lfile$close_all()
+
+# Create a data frame for gene information
+gene_info <- data.frame(
+    gene_id = gene_ids,
+    gene_name = gene_names
+)
+
+# Write the data to 10X format
+
+write10xCounts(
+    path = outdir,
+    x = expr_matrix,
+    gene.id = gene_info$gene_id,
+    gene.symbol = gene_info$gene_name,
+    barcodes = cell_barcodes,
+    version = "3",
+    overwrite = TRUE
+)