Nothing
# ==============================================================================
# filters rows/columns from input matrix `m` until all entries >= `n`,
# such that ea. iteration removes the row/column w/ the smallest summed value.
# ------------------------------------------------------------------------------
.filter_matrix <- function(m, n = 100) {
while (any(m < n)) {
# get candidate rows/cols for removal
i <- m < n
r <- apply(i, 1, any)
c <- apply(i, 2, any)
# get smallest row/col
rs <- rowSums(m)
cs <- colSums(m)
r <- which(r)[which.min(rs[r])]
c <- which(c)[which.min(cs[c])]
# priorities removal of rows over cols
if (rs[r] <= cs[c]) {
m <- m[-r, , drop = FALSE]
} else {
m <- m[, -c, drop = FALSE]
}
if (any(dim(m) == 1))
break
}
return(m)
}
.update_sce <- function(sce) {
# update colData
cd <- as.data.frame(colData(sce))
cd <- mutate_if(cd, is.factor, droplevels)
colData(sce) <- DataFrame(cd, row.names = colnames(sce))
# update metadata
if(!is.null(ei <- metadata(sce)$experiment_info)){
ei <- ei[ei$sample_id %in% levels(sce$sample_id), ]
ei <- mutate_if(ei, is.factor, droplevels)
metadata(sce)$experiment_info <- ei
}
return(sce)
}
.filter_sce <- function(sce, kids, sids) {
cs1 <- sce$cluster_id %in% kids
cs2 <- sce$sample_id %in% sids
sce <- sce[, cs1 & cs2]
sce <- .update_sce(sce)
return(sce)
}
.cluster_colors <- c(
"#DC050C", "#FB8072", "#1965B0", "#7BAFDE", "#882E72",
"#B17BA6", "#FF7F00", "#FDB462", "#E7298A", "#E78AC3",
"#33A02C", "#B2DF8A", "#55A1B1", "#8DD3C7", "#A6761D",
"#E6AB02", "#7570B3", "#BEAED4", "#666666", "#999999",
"#aa8282", "#d4b7b7", "#8600bf", "#ba5ce3", "#808000",
"#aeae5c", "#1e90ff", "#00bfff", "#56ff0d", "#ffff00")
# ==============================================================================
# scale values b/w 0 and 1 using
# low (1%) and high (99%) quantiles as boundaries
# ------------------------------------------------------------------------------
#' @importFrom matrixStats rowQuantiles
.scale <- function(x) {
qs <- rowQuantiles(as.matrix(x), probs = c(.01, .99), na.rm = TRUE)
x <- (x - qs[, 1]) / (qs[, 2] - qs[, 1])
x[x < 0] <- 0
x[x > 1] <- 1
return(x)
}
# ==============================================================================
# wrapper for z-normalization
# ------------------------------------------------------------------------------
.z_norm <- function(x, th = 2.5) {
x <- as.matrix(x)
sds <- rowSds(x, na.rm = TRUE)
sds[sds == 0] <- 1
x <- t(t(x - rowMeans(x, na.rm = TRUE)) / sds)
#x <- (x - rowMeans(x, na.rm = TRUE)) / sds
x[x > th] <- th
x[x < -th] <- -th
return(x)
}
# ------------------------------------------------------------------------------
# generate experimental design metadata table
# for an input SCE or colData data.frame
# ------------------------------------------------------------------------------
#' @importFrom dplyr mutate_at
#' @importFrom methods is
#' @importFrom SummarizedExperiment colData
.make_ei <- function(x) {
if (is(x, "SingleCellExperiment"))
x <- colData(x)
sids <- unique(x$sample_id)
m <- match(sids, x$sample_id)
df <- data.frame(
stringsAsFactors = FALSE,
sample_id = sids,
group_id = x$group_id[m],
n_cells = as.numeric(table(x$sample_id)[sids]))
for (i in c("sample_id", "group_id"))
if (is.factor(x[[i]]))
df <- mutate_at(df, i, factor, levels = levels(x[[i]]))
return(df)
}
# ------------------------------------------------------------------------------
# split cells by cluster-sample
# ------------------------------------------------------------------------------
# x: a SingleCellExperiment or colData
# by: character vector specifying colData column(s) to split by
# > If length(by) == 1, a list of length nlevels(colData$by), else,
# a nested list with 2nd level of length nlevels(colData$by[2])
# ------------------------------------------------------------------------------
#' @importFrom data.table data.table
#' @importFrom purrr map_depth
.split_cells <- function(x,
by = c("cluster_id", "sample_id")) {
if (is(x, "SingleCellExperiment"))
x <- colData(x)
cd <- data.frame(x[by], check.names = FALSE)
cd <- data.table(cd, cell = rownames(x)) %>%
split(by = by, sorted = TRUE, flatten = FALSE)
map_depth(cd, length(by), "cell")
}
# ------------------------------------------------------------------------------
# global p-value adjustment
# ------------------------------------------------------------------------------
# x: results table; a nested list w/
# 1st level = comparisons and 2nd level = clusters
# > adds 'p_adj.glb' column containing globally adjusted p-values
# to the result table of ea. cluster for each comparison
# ------------------------------------------------------------------------------
#' @importFrom purrr map map_depth
#' @importFrom stats p.adjust
.p_adj_global <- function(x) {
names(ks) <- ks <- names(x)
names(cs) <- cs <- names(x[[1]])
lapply(cs, function(c) {
# get p-values
tbl <- map_depth(x, 1, c)
p_val <- map(tbl, "p_val")
# adjust for each comparison
p_adj <- p.adjust(unlist(p_val))
# re-split by cluster
ns <- vapply(p_val, length, numeric(1))
p_adj <- split(p_adj, rep.int(ks, ns))
# insert into results tables
lapply(ks, function(k) {
u <- x[[k]][[c]]
i <- which(colnames(u) == "p_adj.loc")
u[["p_adj.glb"]] <- p_adj[[k]]
u[, c(seq_len(i), ncol(u), seq(i+1, ncol(u)-1))]
})
})
}
# ------------------------------------------------------------------------------
# toy SCE for unit-testing
# ------------------------------------------------------------------------------
#' @importFrom SingleCellExperiment SingleCellExperiment
.toySCE <- function() {
gs <- paste0("gene", seq_len(ngs <- 300))
cs <- paste0("cell", seq_len(ncs <- 2e3))
y <- rnbinom(ngs * ncs, size = 2, mu = 4)
y <- matrix(y, ngs, ncs, TRUE, list(gs, cs))
cd <- mapply(function(i, n)
sample(paste0(i, seq_len(n)), ncs, TRUE),
i = c("k", "s", "g"), n = c(5, 4, 3))
cd <- data.frame(cd, stringsAsFactors = TRUE)
cd$s <- factor(paste(cd$s, cd$g, sep = "."))
colnames(cd) <- paste(c("cluster", "sample", "group"), "id", sep = "_")
SingleCellExperiment(
assay = list(counts = y), colData = cd,
metadata = list(experiment_info = .make_ei(cd)))
}
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