From d47572ec365ca45a17b17e854090e8411806e859 Mon Sep 17 00:00:00 2001 From: "Shao, Xin" Date: Thu, 9 Jun 2022 11:01:29 +0800 Subject: [PATCH] update --- NAMESPACE | 1 + R/methods.R | 4 +- R/plot.R | 62 +++++++++++++++-------- man/plot_cci_lrpairs.Rd | 12 +++-- script/cellchat.R | 37 ++++++++++++++ script/cellphonedb.sh | 109 ++++++++++++++++++++++++++++++++++++++++ vignettes/SpaTalk.pdf | Bin 203167 -> 204018 bytes 7 files changed, 198 insertions(+), 27 deletions(-) create mode 100644 script/cellchat.R create mode 100644 script/cellphonedb.sh diff --git a/NAMESPACE b/NAMESPACE index 4e27d0c..a18fe23 100644 --- a/NAMESPACE +++ b/NAMESPACE @@ -41,6 +41,7 @@ import(ggExtra) import(ggalluvial) import(ggplot2) import(ggrepel) +import(grDevices) import(iterators) import(methods) import(parallel) diff --git a/R/methods.R b/R/methods.R index aa14e74..6132624 100644 --- a/R/methods.R +++ b/R/methods.R @@ -806,10 +806,10 @@ get_lr_path <- function(object, celltype_sender, celltype_receiver, ligand, rece ggi_tf <- unique(pathways[, c("src", "dest", "src_tf", "dest_tf")]) st_data <- .get_st_data(object) st_meta <- .get_st_meta(object) - if (!celltype_sender %in% colnames(object@coef)) { + if (!celltype_sender %in% st_meta$celltype) { stop("Please provide the correct name of celltype_sender!") } - if (!celltype_receiver %in% colnames(object@coef)) { + if (!celltype_receiver %in% st_meta$celltype) { stop("Please provide the correct name of celltype_receiver!") } if (!ligand %in% rownames(st_data)) { diff --git a/R/plot.R b/R/plot.R index c0d3976..6c2676a 100644 --- a/R/plot.R +++ b/R/plot.R @@ -571,13 +571,15 @@ plot_ccdist <- function(object, celltype_sender, celltype_receiver, color = NULL #' @param type Set 'sig' to plot significant LR pairs or set 'number' to plot the number of spatial LR interactions. #' @param fontsize_number fontsize of the numbers displayed in cells. #' @param number_color color of the text. -#' @import ggplot2 pheatmap +#' @param color_low For 'number' type, define the color for the lowest value. +#' @param color_high For 'number' type, define the color for the highest value. +#' @import ggplot2 pheatmap grDevices #' @importFrom ggpubr get_palette #' @importFrom reshape2 dcast #' @export -plot_cci_lrpairs <- function(object, celltype_sender, celltype_receiver, top_lrpairs = 20, color = NULL, - border_color = "black", type = NULL, fontsize_number = 1, number_color = "black") { +plot_cci_lrpairs <- function(object, celltype_sender, celltype_receiver, top_lrpairs = 20, color = NULL, border_color = "black", type = "sig", + fontsize_number = 5, number_color = "black", color_low = NULL, color_high = NULL) { # check if (!is(object, "SpaTalk")) { stop("Invalid class for object: must be 'SpaTalk'!") @@ -624,25 +626,41 @@ plot_cci_lrpairs <- function(object, celltype_sender, celltype_receiver, top_lrp } ligand <- unique(lrpair$ligand) receptor <- unique(lrpair$receptor) - # get lrpair_real - lrpair_real <- object@lr_path$lrpairs - lrpair_real <- lrpair_real[, c(1, 2)] - lrpair_real$score <- 1 - lrpair_mat <- reshape2::dcast(lrpair_real, formula = ligand ~ receptor, fill = 0, value.var = "score") - rownames(lrpair_mat) <- lrpair_mat$ligand - lrpair_mat <- lrpair_mat[, -1] - lrpair_mat <- lrpair_mat[ligand, receptor] - if (!is.data.frame(lrpair_mat)) { - stop("Limited number of ligand-receptor interactions!") - } - plot_res <- matrix("", nrow = length(ligand), ncol = length(receptor)) - rownames(plot_res) <- ligand - colnames(plot_res) <- receptor - for (i in 1:nrow(lrpair)) { - plot_res[lrpair$ligand[i], lrpair$receptor[i]] <- "*" - } - pheatmap::pheatmap(lrpair_mat, cluster_cols = F, cluster_rows = F, color = heat_col, border_color = border_color, - legend = F, display_numbers = plot_res, fontsize_number = fontsize_number, number_color = number_color) + if (type == "sig") { + # get lrpair_real + lrpair_real <- object@lr_path$lrpairs + lrpair_real <- lrpair_real[, c(1, 2)] + lrpair_real$score <- 1 + lrpair_mat <- reshape2::dcast(lrpair_real, formula = ligand ~ receptor, fill = 0, value.var = "score") + rownames(lrpair_mat) <- lrpair_mat$ligand + lrpair_mat <- lrpair_mat[, -1] + lrpair_mat <- lrpair_mat[ligand, receptor] + if (!is.data.frame(lrpair_mat)) { + stop("Limited number of ligand-receptor interactions!") + } + plot_res <- matrix("", nrow = length(ligand), ncol = length(receptor)) + rownames(plot_res) <- ligand + colnames(plot_res) <- receptor + for (i in 1:nrow(lrpair)) { + plot_res[lrpair$ligand[i], lrpair$receptor[i]] <- "*" + } + pheatmap::pheatmap(lrpair_mat, cluster_cols = F, cluster_rows = F, color = heat_col, border_color = border_color, legend = F, display_numbers = plot_res, + fontsize_number = fontsize_number, number_color = number_color, main = "Significantly enriched LRI") + } else { + if (is.null(color_low)) { + color_low <- "orange" + } + if (is.null(color_high)) { + color_high <- "red" + } + lrpair_real <- lrpair[,c("ligand","receptor","lr_co_exp_num")] + lrpair_mat <- reshape2::dcast(lrpair_real, formula = ligand ~ receptor, fill = 0, value.var = "lr_co_exp_num") + rownames(lrpair_mat) <- lrpair_mat$ligand + lrpair_mat <- lrpair_mat[, -1] + heat_color <- grDevices::colorRampPalette(c(color_low, color_high))(max(as.matrix(lrpair_mat))-1) + heat_color <- c("white", heat_color) + pheatmap::pheatmap(lrpair_mat, cluster_cols = F, cluster_rows = F, border_color = border_color, color = heat_color, main = "Number of spatial LRIs") + } } #' @title Plot LR pair diff --git a/man/plot_cci_lrpairs.Rd b/man/plot_cci_lrpairs.Rd index ed8d419..68c20cc 100644 --- a/man/plot_cci_lrpairs.Rd +++ b/man/plot_cci_lrpairs.Rd @@ -11,9 +11,11 @@ plot_cci_lrpairs( top_lrpairs = 20, color = NULL, border_color = "black", - type = NULL, - fontsize_number = 1, - number_color = "black" + type = "sig", + fontsize_number = 5, + number_color = "black", + color_low = NULL, + color_high = NULL ) } \arguments{ @@ -34,6 +36,10 @@ plot_cci_lrpairs( \item{fontsize_number}{fontsize of the numbers displayed in cells.} \item{number_color}{color of the text.} + +\item{color_low}{For 'number' type, define the color for the lowest value.} + +\item{color_high}{For 'number' type, define the color for the highest value.} } \description{ Heatmap with LR pairs of celltype_sender and celltype_receiver diff --git a/script/cellchat.R b/script/cellchat.R new file mode 100644 index 0000000..95f4769 --- /dev/null +++ b/script/cellchat.R @@ -0,0 +1,37 @@ +#============================================== +#================== CellChat ================== +#============================================== +library(CellChat) +library(SpaTalk) +library(tidyverse) +library(Seurat) +options(stringsAsFactors = FALSE) + +load(paste0(system.file(package = 'SpaTalk'),'/extdata/starmap_data.rda')) # starmap_data +load(paste0(system.file(package = 'SpaTalk'),'/extdata/starmap_meta.rda')) # starmap_meta + + +ndata <- starmap_data +celltype <- starmap_meta +rownames(celltype) <- celltype$cell +all(colnames(ndata) == celltype$cell) + +seuratObj <- CreateSeuratObject(counts = ndata) +seuratObj <- NormalizeData(seuratObj) +Idents(seuratObj) <- celltype$celltype + +data.input <- GetAssayData(seuratObj, assay = "RNA", slot = "data") # normalized data matrix +labels <- Idents(seuratObj) +meta <- data.frame(labels = labels, row.names = names(labels)) # create a dataframe of the cell labels + +cellchat <- createCellChat(object = data.input, meta = meta, group.by = "labels") +CellChatDB <- CellChat::CellChatDB.mouse +cellchat@DB <- CellChatDB + +cellchat <- subsetData(cellchat) +future::plan("multisession", workers = 4) # do parallel +cellchat <- identifyOverExpressedGenes(cellchat) +cellchat <- identifyOverExpressedInteractions(cellchat) +cellchat <- computeCommunProb(cellchat) +cellchat <- filterCommunication(cellchat, min.cells = 10) +df.net <- res_cellchat[[use_db]][['starmap']][['starmap']] <- df.net %>% dplyr::select(1:6) diff --git a/script/cellphonedb.sh b/script/cellphonedb.sh new file mode 100644 index 0000000..6636501 --- /dev/null +++ b/script/cellphonedb.sh @@ -0,0 +1,109 @@ +#----------------------------------- +#--- R script after +#----------------------------------- +# Run before python +library(SpaTalk) +library(tidyverse) + +load(paste0(system.file(package = 'SpaTalk'),'/extdata/starmap_data.rda')) # starmap_data +load(paste0(system.file(package = 'SpaTalk'),'/extdata/starmap_meta.rda')) # starmap_meta + +# orthologs gene +ndata <- starmap_data +orthologs_gene <- mapper[rownames(ndata)] +na_gene <- rownames(ndata)[is.na(orthologs_gene)] +ndata <- ndata[!is.na(orthologs_gene),] +rownames(ndata) <- orthologs_gene[!is.na(orthologs_gene)] %>% make.names() + +celltype <- starmap_meta +rownames(celltype) <- celltype$cell +all(colnames(ndata) == celltype$cell) + +# create Seurat object + +seuratObj <- CreateSeuratObject(counts = ndata) +Idents(seuratObj) <- celltype$celltype +seuratObj[['celltype']] <- celltype$celltype +seuratObj <- NormalizeData(seuratObj) + +writeMM(seuratObj@assays$RNA@counts, file = 'data/cpdb_input/starmap_counts_mtx/matrix.mtx') +# save gene and cell names +write(x = rownames(seuratObj@assays$RNA@counts), file = "data/cpdb_input/starmap_counts_mtx/features.tsv") +write(x = colnames(seuratObj@assays$RNA@counts), file = "data/cpdb_input/starmap_counts_mtx/barcodes.tsv") + +seuratObj@meta.data$Cell = rownames(seuratObj@meta.data) +df = seuratObj@meta.data[, c('Cell', 'celltype')] +write.table(df, file ='data/cpdb_input/starmap_meta.tsv', sep = '\t', quote = F, row.names = F) + +#=============================================================================================== + + +#----------------------------------- +#--- python script after +#----------------------------------- + +conda activate cpdb +cd data/cpdb_input/ +cellphonedb method statistical_analysis starmap_meta.tsv starmap_counts_mtx/ --counts-data gene_name + +#----------------------------------- +#--- python script before +#----------------------------------- + + +#================================================================================= + +#----------------------------------- +#--- R script after +#----------------------------------- + +# standardize cpdb output +res_cpdb_uniondb <- list() +# create mapper to convert gene into mouse gene +mapper <- gene2orthologs$mousegene +names(mapper) <- gene2orthologs$humangene +gene2orthologs <- readRDS("~/待处理/Spatalk_Benchmark/verified_database/cpdb/gene2orthologs_human_mouse.rds") + +cpdb2df <- function(data) { + df_data <- data.frame() + Ligand = c() + Receptor = c() + Ligand.Cluster = c() + Receptor.Cluster = c() + isReceptor_fst = c() + isReceptor_scn = c() + MeanLR = c() + + for (i in 1:nrow(data)) { + pair <- strsplit(data$`interacting_pair`[i],split = "_") + for (j in 13:ncol(data)) { + c_pair = strsplit(colnames(data)[j],"\\|") + if (data[i,j] != 0.0) { + Ligand <- c(Ligand,pair[[1]][1]) + Receptor <- c(Receptor,pair[[1]][2]) + Ligand.Cluster <- c(Ligand.Cluster,c_pair[[1]][1]) + Receptor.Cluster <- c(Receptor.Cluster,c_pair[[1]][2]) + isReceptor_fst <- c(isReceptor_fst,data$`receptor_a`[i]) + isReceptor_scn <- c(isReceptor_scn,data$`receptor_b`[i]) + MeanLR <- c(MeanLR,data[i,j]) + } + + } + + } + df_data <- data.frame(Ligand = Ligand, Receptor = Receptor, + Ligand.Cluster = Ligand.Cluster,Receptor.Cluster = Receptor.Cluster, + isReceptor_fst = isReceptor_fst, + isReceptor_scn = isReceptor_scn, + MeanLR = MeanLR) + return(df_data) +} + +#--- starmap +cpdb_O <- read.table(paste0("cpdb_input/","/out/significant_means.txt"),sep = "\t",header = T,check.names = F,stringsAsFactors = F) +cpdb_O[is.na(cpdb_O)] <- 0 +cpdb_O <- cpdb2df(cpdb_O) + +cpdb_O <- cpdb_O %>% mutate(Ligand = mapper[Ligand], Receptor = mapper[Receptor]) +cpdb_O <- cpdb_O %>% rename(receptor = Ligand,ligand = Receptor, sender = Receptor.Cluster, receiver = Ligand.Cluster) + diff --git a/vignettes/SpaTalk.pdf b/vignettes/SpaTalk.pdf index 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