R/apl.R
In ClemensKohl/APL: Association Plots
Defines functions
run_APL
apl
apl_plotly
apl_ggplot
apl_topGO
apl_score
apl_coords
Documented in
apl
apl_coords
apl_ggplot
apl_plotly
apl_score
apl_topGO
run_APL
#' @include constructor.R
NULL
#' Calculate Association Plot coordinates
#'
#' @description
#' Calculates the Association Plot coordinates for either the rows,
#' columns or both (default).
#'
#' @details
#' Coordinates (x,y) of row vector \eqn{\vec{r}} are defined as
#' \deqn{x(\vec{r}) := \left|\vec{r}\right|\cos(\phi(\vec{r}))}
#' \deqn{y(\vec{r}) := \left|\vec{r}\right|\sin(\phi(\vec{r}))}
#' The x-direction is determined by calculating the centroid of the columns
#' selected with the indices in "group".
#'
#' @return
#' Returns input "cacomp" object and adds components "apl_rows" and/or
#' "apl_cols" for row and column coordinates.
#' In "group" the indices of the columns used to calculate the
#' centroid are saved.
#'
#' @references
#' Association Plots: Visualizing associations in high-dimensional
#' correspondence analysis biplots
#' Elzbieta Gralinska, Martin Vingron
#' bioRxiv 2020.10.23.352096; doi: https://doi.org/10.1101/2020.10.23.352096
#'
#' @param caobj A "cacomp" object with principal row coordinates and
#' standardized column coordinates calculated.
#' @param group Numeric/Character. Vector of indices or column names of
#' the columns to calculate centroid/x-axis direction.
#' @param calc_rows TRUE/FALSE. Whether apl row coordinates should
#' be calculated. Default TRUE.
#' @param calc_cols TRUE/FALSE. Whether apl column coordinates should
#' be calculated. Default TRUE.
#' @export
#'
#' @examples
#' set.seed(1234)
#' # Simulate scRNAseq data
#' cnts <- data.frame(cell_1 = rpois(10, 5),
#' cell_2 = rpois(10, 10),
#' cell_3 = rpois(10, 20),
#' cell_4 = rpois(10, 20))
#' rownames(cnts) <- paste0("gene_", 1:10)
#' cnts <- as.matrix(cnts)
#'
#' # Run correspondence analysis
#' ca <- cacomp(obj = cnts, princ_coords = 3)
#' # Calculate APL coordinates
#' ca <- apl_coords(ca, group = 3:4)
apl_coords <- function(caobj, group, calc_rows = TRUE, calc_cols = TRUE) {
stopifnot(is(caobj, "cacomp"))
rows <- caobj@prin_coords_rows
cols <- caobj@std_coords_cols
cent <- cols
if (is(group, "numeric")){
subgroup <- cent[group,]
} else if (is(group, "character")){
idx <- match(group, rownames(cent))
idx <- na.omit(idx)
group <- idx
subgroup <- cent[idx, ]
if (anyNA(idx)){
warning("Not all names in 'group' are contained in the column names. ",
"Non-matching values were ignored.")
}
} else {
stop("Parameter group has to be either of type 'numeric' or 'character'.")
}
if (length(group) == 1){
avg_group_coords <- subgroup # single sample
} else {
avg_group_coords <- colMeans(subgroup) # centroid vector.
}
length_vector_group <- sqrt(drop(avg_group_coords %*% avg_group_coords))
length_vector_rows <- sqrt(Matrix::rowSums(rows^2))
length_vector_cols <- sqrt(Matrix::rowSums(cols^2))
if (calc_rows == TRUE){
# message("Calculating APL row coordinates ...")
# r⋅X = |r|*|X|*cosθ
# x(r) = (r⋅X)/|X| = |r|*cosθ
rowx <- drop(rows %*% avg_group_coords)/length_vector_group
# pythagoras, y(r)=b²=c²-a²
rowy <- sqrt(length_vector_rows^2 - rowx^2)
rowx[is.na(rowx)] <- 0
rowy[is.na(rowy)] <- 0
# rowx[is.infinite(rowx)] <- 0
# rowy[is.infinite(rowy)] <- 0
caobj@apl_rows <- cbind("x"=rowx, "y"=rowy)
}
if (calc_cols == TRUE){
# message("Calculating APL column coordinates ...")
colx <- drop(cols %*% avg_group_coords)/length_vector_group
coly <- sqrt(length_vector_cols^2 - colx^2)
colx[is.na(colx)] <- 0
coly[is.na(coly)] <- 0
# colx[is.infinite(colx)] <- 0
# coly[is.infinite(coly)] <- 0
caobj@apl_cols <- cbind("x"=colx, "y"=coly)
}
if (is(group, "numeric")){
caobj@group <- group
} else if (is(group, "character")){
idx <- match(group, rownames(cols))
idx <- na.omit(idx)
caobj@group <- idx
}
stopifnot(validObject(caobj))
return(caobj)
}
#' Find rows most highly associated with a condition
#'
#' @description
#' Ranks rows by a calculated score which balances the association of the row
#' with the condition and how associated it is with other conditions.
#'
#' @details
#' The score is calculated by permuting the values of each row to determine the
#' cutoff angle of the 99% quantile.
#' \deqn{S_{alpha}(x,y)=x-\frac{y}{\tan\alpha}}
#' By default the permutation is repeated 10 times, but for very large matrices
#' this can be reduced.
#' If store_perm is TRUE the permuted data is stored in the cacomp object and
#' can be used for future scoring.
#' @return
#' Returns the input "cacomp" object with "APL_score" component added.
#' APL_score contains a data frame with ranked rows, their score and their
#' original row number.
#'
#' @references
#' Association Plots: Visualizing associations in high-dimensional
#' correspondence analysis biplots \cr
#' Elzbieta Gralinska, Martin Vingron \cr
#' bioRxiv 2020.10.23.352096; doi: https://doi.org/10.1101/2020.10.23.352096
#'
#' @param caobj A "cacomp" object with principal row coordinates and
#' standardized column coordinates calculated.
#' @param mat A numeric matrix. For sequencing a count matrix, gene expression
#' values with genes in rows and samples/cells in columns.
#' Should contain row and column names.
#' @param dims Integer. Number of CA dimensions to retain. Needs to be the same
#' as in caobj!
#' @param group Vector of indices of the columns to calculate centroid/x-axis
#' direction.
#' @param reps Integer. Number of permutations to perform. Default = 10.
#' @param quant Numeric. Single number between 0 and 1 indicating the quantile
#' used to calculate the cutoff. Default 0.99.
#' @param python A logical value indicating whether to use singular-value
#' decomposition from the python package torch.
#' @param store_perm Logical. Whether permuted data should be stored in the CA
#' object.
#' This implementation dramatically speeds up computation compared to `svd()`
#' in R.
#' @export
#'
#' @examples
#' set.seed(1234)
#'
#' # Simulate counts
#' cnts <- mapply(function(x){rpois(n = 500, lambda = x)},
#' x = sample(1:20, 50, replace = TRUE))
#' rownames(cnts) <- paste0("gene_", 1:nrow(cnts))
#' colnames(cnts) <- paste0("cell_", 1:ncol(cnts))
#'
#' # Run correspondence analysis.
#' ca <- cacomp(obj = cnts, princ_coords = 3)
#'
#' # Calculate APL coordinates:
#' ca <- apl_coords(ca, group = 1:10)
#'
#' # Rank genes by S-alpha score
#' ca <- apl_score(ca, mat = cnts)
apl_score <- function(caobj,
mat,
dims = caobj@dims,
group = caobj@group,
reps=10,
quant = 0.99,
python = FALSE,
store_perm = TRUE){
if (!is(caobj,"cacomp")){
stop("Not a CA object. Please run cacomp() and apl_coords() first!")
}
if (is.empty(caobj@apl_rows)){
stop("Please run apl_coords() first!")
}
names <- colnames(mat)
row_num <- nrow(caobj@apl_rows)
margin <- 1
pc <- 1
cc <- FALSE
cr <- TRUE
if (is(group, "character")){
idx <- match(group, names)
idx <- na.omit(idx)
group <- idx
}
if (caobj@dims == 1 && !is.empty(caobj@dims)){
row_num <- 1
}
apl_perm <- data.frame("x" = rep(0, row_num*reps),
"y" = rep(0, row_num*reps))
saved_ca <- list()
pb <- txtProgressBar(min = 0, max = reps, style = 3)
for (k in seq(reps)){
#permute rows and rerun cacomp
if(isTRUE(store_perm) & identical(reps, attr(caobj@permuted_data,'reps'))){
calist <- caobj@permuted_data[[k]][seq_len(3)]
mat <- caobj@permuted_data[[k]]$mat
mat <- mat[rownames(mat) %in% rownames(calist$std_coords_rows),]
caobjp <- recompute(calist, mat)
} else {
mat_perm <- t(apply(mat, margin, FUN=sample))
colnames(mat_perm) <- colnames(mat)
suppressWarnings(caobjp <- cacomp(obj = mat_perm,
python = python,
coords = TRUE,
princ_coords = pc,
dims = dims,
top = caobj@top_rows,
inertia = FALSE))
if(isTRUE(store_perm)){
x <- list("std_coords_cols" = caobjp@std_coords_cols,
"std_coords_rows" = caobjp@std_coords_rows,
"D" = caobjp@D,
"mat" = mat_perm)
saved_ca[[k]] <- x
}
}
caobjp <- apl_coords(caobj = caobjp,
group = group,
calc_cols = cc,
calc_rows = cr)
idx <- ((seq_len(row_num)+((k-1)*row_num)))
apl_perm[idx,] <- caobjp@apl_rows
setTxtProgressBar(pb, k)
}
close(pb)
apl_perm[,3] <- apl_perm[,1]/apl_perm[,2] # cotan between row and x axis
apl_perm[,3][is.na(apl_perm[,3])] <- 0
cutoff_cotan <- quantile(apl_perm[,3], quant)
score <- caobj@apl_rows[,1] - (caobj@apl_rows[,2] * cutoff_cotan)
ranking <- data.frame("Rowname" = rownames(caobj@apl_rows),
"Score" = score,
"Row_num" = seq_len(nrow(caobj@apl_rows)))
ranking <- ranking[order(ranking$Score, decreasing = TRUE),]
ranking$Rank <- seq_len(nrow(ranking))
caobj@APL_score <- ranking
if(isTRUE(store_perm) & !identical(reps, attr(caobj@permuted_data,'reps'))){
caobj@permuted_data <- saved_ca
attr(caobj@permuted_data,'cutoff') <- cutoff_cotan
attr(caobj@permuted_data,'reps') <- reps
}
stopifnot(validObject(caobj))
return(caobj)
}
#' Run Gene overrepresentation analysis with topGO
#'
#' @description
#' This function uses the Kolmogorov-Smirnov test as implemented by the package
#' topGO to test for overrepresentation in Gene Ontology gene sets.
#'
#' @details
#' For a chosen group of cells/samples,
#' the top 'ngenes' group specific genes are used for gene overrepresentation
#' analysis.
#' The genes are ranked either by the precomputed APL score, or, if
#' not available by their APL x-coordinates.
#'
#' @return
#' A data.frame containing the gene sets with the highest overrepresentation.
#'
#' @references
#' Adrian Alexa and Jorg Rahnenfuhrer \cr
#' topGO: Enrichment Analysis for Gene Ontology. \cr
#' R package version 2.42.0.
#'
#' @param caobj A "cacomp" object with principal row coordinates and
#' standardized column coordinates calculated.
#' @param ontology Character string. Chooses GO sets for 'BP'
#' (biological processes), 'CC' (cell compartment) or 'MF' (molecular function).
#' @param organism Character string. Either 'hs' (homo sapiens), 'mm'
#' (mus musculus) or the name of the organism package such as 'org.*.eg.db'.
#' @param ngenes Numeric. Number of top ranked genes to test for
#' overrepresentation.
#' @param score_cutoff numeric. S-alpha score cutoff. Only genes with a score
#' larger will be tested.
#' @param use_coords Logical. Whether the x-coordinates of the row APL
#' coordinates should be used for ranking.
#' Only recommended when no S-alpha score (see apl_score()) can be calculated.
#' @param return_plot Logical. Whether a plot of significant gene sets should
#' be additionally returned.
#' @param top_res Numeric. Number of top scoring genes to plot.
#'
#' @export
#' @examples
#' library(Seurat)
#' set.seed(1234)
#' cnts <- GetAssayData(pbmc_small, assay = "RNA", slot = "counts")
#' cnts <- as.matrix(cnts)
#'
#' # Run CA on example from Seurat
#'
#' ca <- cacomp(pbmc_small,
#' princ_coords = 3,
#' return_input = FALSE,
#' assay = "RNA",
#' slot = "counts")
#'
#' grp <- which(Idents(pbmc_small) == 2)
#' ca <- apl_coords(ca, group = grp)
#' ca <- apl_score(ca,
#' mat = cnts)
#'
#' enr <- apl_topGO(ca,
#' ontology = "BP",
#' organism = "hs")
#'
#' plot_enrichment(enr)
apl_topGO <- function(caobj,
ontology,
organism="hs",
ngenes = 1000,
score_cutoff = 0,
use_coords = FALSE,
return_plot = FALSE,
top_res = 15){
topGO::groupGOTerms()
if(!is.empty(caobj@APL_score) & !isTRUE(use_coords)){
Score_ord <- caobj@APL_score[order(caobj@APL_score$Score, decreasing=TRUE),]
sel <- which(!Score_ord$Score >= score_cutoff)[1]
sel <- sel-1
ranked_genes <- seq_len(nrow(Score_ord))
names(ranked_genes) <- Score_ord$Rowname
} else if (isTRUE(use_coords) & is.empty(caobj@APL_score)) {
if(is.empty(caobj@apl_rows)){
stop("No APL coordinates found for rows. Please first run apl_coords.\n")
}
if (ngenes > nrow(caobj@apl_rows)){
stop("ngenes is larger than the total number of genes.\n")
} else if (ngenes == nrow(caobj@apl_rows)){
warning("You have selected all available genes.\n")
}
ranked_genes <- seq_len(nrow(caobj@apl_rows))
names(ranked_genes) <- rownames(caobj@apl_rows)[order(caobj@apl_rows[,1],
decreasing = TRUE)]
sel <- ngenes
} else {
stop("APL scores not present but use_coords set to FALSE.\n")
}
if (organism == "hs"){
organism <- "org.Hs.eg.db"
} else if (organism == "mm"){
organism <- "org.Mm.eg.db"
} else {
warning("Custom organism chosen.\n")
organism <- organism
}
if (!ontology %in% c("BP", "CC", "MF")){
stop("Please choose one of the following ontologies: 'BP', 'CC' or 'MF'.\n")
}
gene_sets <- topGO::annFUN.org(whichOnto=ontology,
feasibleGenes=NULL,
mapping=organism,
ID="symbol")
GOdata <- new("topGOdata",
ontology = ontology,
allGenes = ranked_genes,
annot = topGO::annFUN.GO2genes,
GO2genes = gene_sets,
geneSelectionFun = function(x) x<sel,
nodeSize=5)
results_test <- topGO::runTest(GOdata,
algorithm = "elim",
statistic = "fisher")
goEnrichment <- topGO::GenTable(GOdata,
raw.p.value = results_test,
topNodes = length(results_test@score),
numChar = 1200)
if (isTRUE(return_plot)){
if(top_res > length(results_test@score)){
warning("More top nodes selected via top_res than available. ",
"Returning max. number of nodes instead.\n")
top_res <- min(top_res, length(results_test@score))
}
topGO::showSigOfNodes(GOdata,
score(results_test),
firstSigNodes = top_res,
useInfo = 'def')
}
return(goEnrichment)
}
#'
#' Plot Association Plot with ggplot
#' @description
#' Uses ggplot to plot an Association Plot
#'
#' @param rows Row APL-coordinates
#' @param rows_group Row AP-coordinates to highlight
#' @param rows_scored Row AP-coordinates of rows above a score cutoff.
#' @param cols Column AP-coordinates
#' @param cols_group Column AP-coordinates for the group to be highlighted.
#' @param rows_color Color for rows
#' @param rows_high_color Color for rows to be highlighted.
#' @param cols_color Column points color.
#' @param cols_high_color Color for column points to be highlighted..
#' @param score_color Color scheme for row points with a score.
#' @param row_labs Logical. Whether labels for rows indicated by rows_idx
#' should be labeled with text. Default TRUE.
#' @param col_labs Logical. Whether labels for columns indicated by cols_idx
#' shouls be labeled with text. Default FALSE.
#' @param show_score Logical. Whether the S-alpha score should be shown in
#' the plot.
#' @param show_cols Logical. Whether column points should be plotted.
#' @param show_rows Logical. Whether row points should be plotted.
#'
#' @returns
#' ggplot Association Plot
apl_ggplot <- function(rows,
rows_group = NULL,
cols,
cols_group = NULL,
rows_scored = NULL,
rows_color = "#0066FF",
rows_high_color = "#FF0000",
cols_color = "#601A4A",
cols_high_color = "#EE442F",
score_color = "rainbow",
row_labs = FALSE,
col_labs = FALSE,
show_score = FALSE,
show_cols = FALSE,
show_rows = TRUE){
p <- ggplot2::ggplot()
if (isTRUE(show_rows)){
p <- p +
ggplot2::geom_point(data=rows,
ggplot2::aes(x=.data$x, y=.data$y),
color = rows_color,
alpha = 0.7,
shape = 1) #16 point, 1 circle.
if (isTRUE(show_score)){
p <- p +
ggplot2::geom_point(data=rows_scored,
ggplot2::aes(x=.data$x,
y=.data$y,
fill = .data$Score),
alpha = 0.7,
shape = 21,
stroke = 0.5)
if (score_color == "rainbow"){
hex <- c("#00007F",
"blue",
"#007FFF",
"cyan",
"#7FFF7F",
"yellow",
"#FF7F00",
"red",
"#7F0000")
p <- p +
ggplot2::scale_fill_gradientn(colours = hex,
name = expression(S*alpha))
} else if (score_color == "viridis"){
p <- p +
ggplot2::scale_color_viridis_c(option = "D")
}
}
if (!is.null(rows_group)){
p <- p +
ggplot2::geom_point(data=rows_group,
ggplot2::aes(x=.data$x, y=.data$y),
color = rows_high_color,
shape = 19)
if(isTRUE(row_labs)){
p <- p +
ggrepel::geom_text_repel(data = rows_group,
ggplot2::aes(x=.data$x,
y=.data$y,
label=.data$rownms),
color = rows_high_color,
max.overlaps = Inf)
}
}
}
if (isTRUE(show_cols)){
p <- p +
ggplot2::geom_point(data=cols,
ggplot2::aes(x=.data$x, y=.data$y),
color = cols_color,
shape = 4)
if (!is.null(cols_group)){
p <- p +
ggplot2::geom_point(data=cols_group,
ggplot2::aes(x=.data$x, y=.data$y),
color = cols_high_color,
shape = 4)
if(col_labs == TRUE){
p <- p +
ggrepel::geom_text_repel(data=cols_group,
ggplot2::aes(x=.data$x,
y=.data$y,
label=.data$rownms),
color = cols_high_color)
}
}
}
p <- p +
ggplot2::labs(title="Association Plot") +
ggplot2::theme_bw()
return(p)
}
#'
#' Plot Association Plot with plotly
#' @description
#' Uses plotly to generate an interactive Association Plot
#'
#' @param rows Row APL-coordinates
#' @param rows_group Row AP-coordinates to highlight
#' @param rows_scored Row AP-coordinates of rows above a score cutoff.
#' @param cols Column AP-coordinates
#' @param cols_group Column AP-coordinates for the group to be highlighted.
#' @param rows_color Color for rows
#' @param rows_high_color Color for rows to be highlighted.
#' @param cols_color Column points color.
#' @param cols_high_color Color for column points to be highlighted.
#' @param score_color Color scheme for row points with a score.
#' @param row_labs Logical. Whether labels for rows indicated by rows_idx
#' should be labeled with text. Default TRUE.
#' @param col_labs Logical. Whether labels for columns indicated by cols_idx
#' shouls be labeled with text. Default FALSE.
#' @param show_score Logical. Whether the S-alpha score should be shown in
#' the plot.
#' @param show_cols Logical. Whether column points should be plotted.
#' @param show_rows Logical. Whether row points should be plotted.
#'
#' @returns
#' Interactive plotly Association Plot
apl_plotly <- function(rows,
rows_group =NULL,
cols,
cols_group,
rows_scored = NULL,
rows_color = "#0066FF",
rows_high_color = "#FF0000",
cols_color = "#601A4A",
cols_high_color = "#EE442F",
score_color = "rainbow",
row_labs = FALSE,
col_labs = FALSE,
show_score = FALSE,
show_cols = FALSE,
show_rows = TRUE){
if (row_labs == FALSE){
rlabs <- 'markers'
rowfont <- NULL
} else {
rlabs <- 'markers+text'
rowfont <- list(color=rows_high_color)
}
if (col_labs == FALSE){
clabs <- 'markers'
colfont <- NULL
} else {
clabs <- 'markers+text'
colfont <- list(color = cols_high_color)
}
p <- plotly::plot_ly()
if(isTRUE(show_rows)){
color_fix <- rows_color
colors_fun <- NULL
color_bar <- NULL
sym <- "circle-open"
p <- p %>%
plotly::add_trace(data = rows,
x = ~x,
y = ~y,
mode = 'markers',
text = rows$rownms,
opacity = 0.7,
textposition = "left",
marker = list(color = color_fix,
colorscale = colors_fun,
symbol = sym,
colorbar = color_bar,
size = 5),
name = 'genes',
hoverinfo = 'text',
type = 'scatter')
if (isTRUE(show_score)){
if (score_color == "rainbow"){
colors_fun <- "Jet"
} else if (score_color == "viridis"){
colors_fun <- 'Viridis'
}
color_fix <- as.formula("~Score")
color_bar <- list(title = "S\u03B1", len=0.5)
sym <- "circle"
p <- p %>%
plotly::add_trace(data = rows_scored,
x = ~x,
y = ~y,
mode = 'markers',
text = rows_scored$rownms,
opacity = 0.7,
textposition = "left",
marker = list(color = color_fix,
colorscale = colors_fun,
symbol = sym,
colorbar = color_bar,
size = 5,
line = list(color = '#000000', #black
width = 0.5)),
name = 'genes (scored)',
hoverinfo = 'text',
type = 'scatter')
}
if (!is.null(rows_group)){
p <- p %>%
plotly::add_trace(data = rows_group,
x = ~x,
y = ~y,
mode = rlabs,
text = rows_group$rownms,
textposition = "left",
textfont=rowfont,
marker = list(symbol = 'circle',
color = rows_high_color,
size = 5),
name = 'marked genes',
hoverinfo = 'text',
type = 'scatter')
}
}
if(isTRUE(show_cols)){
p <- p %>%
plotly::add_trace(data=cols,
x = ~x,
y = ~y,
mode = 'markers',
text = cols$rownms,
textposition = "left",
marker = list(color = cols_color,
symbol = 'x',
size = 5),
name = 'samples',
hoverinfo = 'text',
type = 'scatter')
if (!is.null(cols_group)){
p <- p %>% plotly::add_trace(data = cols_group,
x = ~x,
y = ~y,
mode = clabs,
text = cols_group$rownms,
textposition = "left",
textfont=colfont,
marker = list(symbol = 'x',
color = cols_high_color,
size = 5),
name = 'marked samples',
hoverinfo = 'text',
type = 'scatter')
}
}
p <- p %>%
plotly::layout(title = paste('Association Plot'),
xaxis = list(title = 'Distance from origin (x)',
rangemode = "tozero"),
yaxis = list(title = 'Distance from gene to sample line (y)',
rangemode = "tozero"),
showlegend = TRUE)
return(p)
}
#' Association Plot
#'
#' @description
#' Plot an Association Plot for the chosen columns.
#'
#' @details
#' For an interactive plot type="plotly" can be chosen, otherwise a static plot
#' will be returned.
#' The row and column coordinates have to be already calculated by
#' `apl_coords()`.
#'
#' @return
#' Either a ggplot or plotly object.
#'
#' @references
#' Association Plots: Visualizing associations in high-dimensional
#' correspondence analysis biplots \cr
#' Elzbieta Gralinska, Martin Vingron \cr
#' bioRxiv 2020.10.23.352096; doi: https://doi.org/10.1101/2020.10.23.352096 \cr
#'
#' @param caobj An object of class "cacomp" and "APL" with apl
#' coordinates calculated.
#' @param type "ggplot"/"plotly". For a static plot a string "ggplot",
#' for an interactive plot "plotly". Default "ggplot".
#' @param rows_idx numeric/character vector.
#' Indices or names of the rows that should be labelled. Default NULL.
#' @param cols_idx numeric/character vector.
#' Indices or names of the columns that should be labelled.
#' Default is only to label columns making up the centroid: caobj@group.
#' @param row_labs Logical. Whether labels for rows indicated by rows_idx
#' should be labeled with text. Default TRUE.
#' @param col_labs Logical. Whether labels for columns indicated by cols_idx
#' shouls be labeled with text. Default FALSE.
#' @param show_score Logical. Whether the S-alpha score should be shown in
#' the plot.
#' @param show_cols Logical. Whether column points should be plotted.
#' @param show_rows Logical. Whether row points should be plotted.
#' @param score_cutoff Numeric. Rows (genes) with a score >= score_cutoff will
#' be colored according to their score if show_score = TRUE.
#' @param score_color Either "rainbow" or "viridis".
#' @export
#' @examples
#' set.seed(1234)
#'
#' # Simulate counts
#' cnts <- mapply(function(x){rpois(n = 500, lambda = x)},
#' x = sample(1:100, 50, replace = TRUE))
#' rownames(cnts) <- paste0("gene_", 1:nrow(cnts))
#' colnames(cnts) <- paste0("cell_", 1:ncol(cnts))
#'
#' # Run correspondence analysis
#' ca <- cacomp(obj = cnts, princ_coords = 3)
#'
#' # Calculate APL coordinates for arbitrary group
#' ca <- apl_coords(ca, group = 1:10)
#'
#' # plot results
#' # Note:
#' # Due to random gene expression & group, no highly
#' # associated genes are visible.
#' apl(ca, type = "ggplot")
apl <- function(caobj,
type="ggplot",
rows_idx = NULL,
cols_idx = caobj@group,
row_labs = FALSE,
col_labs = FALSE,
show_score = FALSE,
show_cols = FALSE,
show_rows = TRUE,
score_cutoff = 0,
score_color = "rainbow"){
if (!is(caobj,"cacomp")){
stop("Not a CA object. Please run cacomp() and apl_coords() first!")
}
if (is.empty(caobj@apl_rows) || is.empty(caobj@apl_cols)){
stop("Please run apl_coords() first!")
}
if (is(rows_idx, "character")){
names <- rownames(caobj@apl_rows)
idx <- match(rows_idx, names)
idx <- na.omit(idx)
rows_idx <- idx
}
if (is(cols_idx, "character")){
names <- rownames(caobj@apl_cols)
idx <- match(cols_idx, names)
idx <- na.omit(idx)
cols_idx <- idx
}
if (is.numeric(cols_idx)){
group_cols <- data.frame(rownms = rownames(caobj@apl_cols)[cols_idx],
x = caobj@apl_cols[cols_idx,"x"],
y = caobj@apl_cols[cols_idx,"y"],
row.names = NULL)
} else {
group_cols <- NULL
}
if(is.numeric(rows_idx)){
group_rows <- data.frame(rownms = rownames(caobj@apl_rows)[rows_idx],
x = caobj@apl_rows[rows_idx,"x"],
y = caobj@apl_rows[rows_idx,"y"],
row.names = NULL)
} else {
group_rows <- NULL
}
rows_color <- "#0066FF"
rows_high_color <- "#FF0000"
cols_color <- "#601A4A"
cols_high_color <- "#EE442F"
apl_rows.tmp <- data.frame(rownms = rownames(caobj@apl_rows),
caobj@apl_rows)
apl_cols.tmp <- data.frame(rownms = rownames(caobj@apl_cols),
caobj@apl_cols)
if (isTRUE(show_score)) {
apl_scores <- caobj@APL_score$Score[order(caobj@APL_score$Row_num)]
apl_rows.tmp$Score <- apl_scores
idx <- which(apl_scores >= score_cutoff)
if (is.empty(idx)) {
show_score <- FALSE
warning("No rows with a Score >= score_cutoff of ",
score_cutoff,
". Choose lower cutoff.")
} else {
apl_scored.tmp <- apl_rows.tmp[idx,]
apl_rows.tmp <- apl_rows.tmp[-idx,]
}
}
if (type == "ggplot"){
p <- apl_ggplot(rows = apl_rows.tmp,
rows_group = group_rows,
cols = apl_cols.tmp,
cols_group = group_cols,
rows_scored = apl_scored.tmp,
rows_color = rows_color,
rows_high_color = rows_high_color,
cols_color = cols_color,
cols_high_color = cols_high_color,
score_color = score_color,
row_labs = row_labs,
col_labs = col_labs,
show_score = show_score,
show_cols = show_cols,
show_rows = show_rows)
return(p)
} else if (type == "plotly"){
p <- apl_plotly(rows = apl_rows.tmp,
rows_group = group_rows,
cols = apl_cols.tmp,
cols_group = group_cols,
rows_scored = apl_scored.tmp,
rows_color = rows_color,
rows_high_color = rows_high_color,
cols_color = cols_color,
cols_high_color = cols_high_color,
score_color = score_color,
row_labs = row_labs,
col_labs = col_labs,
show_score = show_score,
show_cols = show_cols,
show_rows = show_rows)
return(p)
} else {
stop("Please specify plot = \"ggplot\" or \"plotly\".",
" Other options are not accepted.")
}
}
#' Internal function to compute and plot Association Plot
#'
#' @description
#' Computes singular value decomposition and coordinates for
#' the Association Plot.
#'
#' @details
#' The function is a wrapper that calls `cacomp()`, `apl_coords()`,
#' `apl_score()` and finally `apl()` for ease of use.
#' The chosen defaults are most useful for genomics experiments, but for more
#' fine grained control the functions
#' can be also run individually for the same results.
#' If score = FALSE, nrow and reps are ignored. If mark_rows is not NULL score
#' is treated as if FALSE.
#' @return
#' Association Plot (plotly object).
#'
#' @references
#' Association Plots: Visualizing associations in high-dimensional
#' correspondence analysis biplots \cr
#' Elzbieta Gralinska, Martin Vingron \cr
#' bioRxiv 2020.10.23.352096; doi: https://doi.org/10.1101/2020.10.23.352096 \cr
#'
#' @param obj A numeric matrix. For sequencing usually a count matrix,
#' gene expression values with genes in rows and samples/cells in columns.
#' Should contain row and column names.
#'
#' @param caobj A "cacomp" object as outputted from `cacomp()`. If not supplied
#' will be calculated. Default NULL.
#'
#' @param dims Integer. Number of dimensions to keep. Default NULL (keeps all
#' dimensions).
#'
#' @param group Numeric/Character. Vector of indices or column names of the
#' columns to calculate centroid/x-axis direction.
#'
#' @param nrow Integer. The top nrow scored row labels will be added to the
#' plot if score = TRUE. Default 10.
#'
#' @param top Integer. Number of most variable rows to retain. Default 5000
#' rows (set NULL to keep all).
#'
#' @param score Logical. Whether rows should be scored and ranked. Ignored when
#' a vector is supplied to mark_rows. Default TRUE.
#'
#' @param mark_rows Character vector. Names of rows that should be highlighted
#' in the plot. If not NULL, score is ignored. Default NULL.
#'
#' @param mark_cols Character vector. Names of cols that should be highlighted
#' in the plot.
#'
#' @param reps Integer. Number of permutations during scoring. Default 3.
#'
#' @param python A logical value indicating whether to use singular value
#' decomposition from the python package torch.
#' This implementation dramatically speeds up computation compared to `svd()`
#' in R.
#'
#' @param row_labs Logical. Whether labels for rows indicated by rows_idx
#' should be labeled with text. Default TRUE.
#'
#' @param col_labs Logical. Whether labels for columns indicated by cols_idx
#' should be labeled with text. Default TRUE.
#'
#' @param type "ggplot"/"plotly". For a static plot a string "ggplot", for an
#' interactive plot "plotly". Default "plotly".
#'
#' @param show_cols Logical. Whether column points should be plotted.
#'
#' @param show_rows Logical. Whether row points should be plotted.
#'
#' @param score_cutoff Numeric. Rows (genes) with a score >= score_cutoff
#' will be colored according to their score if show_score = TRUE.
#'
#' @param score_color Either "rainbow" or "viridis".
#'
#' @param pd_method Which method to use for pick_dims (\link[APL]{pick_dims}).
#'
#' @param pd_reps Number of repetitions performed when using "elbow_rule" in
#' `pick_dims`.
#' (\link[APL]{pick_dims})
#'
#' @param pd_use Whether to use `pick_dims` (\link[APL]{pick_dims}) to determine
#' the number of dimensions. Ignored when `dims` is set by the user.
#'
#' @examples
#' set.seed(1234)
#'
#' # Simulate counts
#' cnts <- mapply(function(x){rpois(n = 500, lambda = x)},
#' x = sample(1:100, 50, replace = TRUE))
#' rownames(cnts) <- paste0("gene_", 1:nrow(cnts))
#' colnames(cnts) <- paste0("cell_", 1:ncol(cnts))
#'
#' # (nonsensical) APL
#' APL:::run_APL(obj = cnts,
#' group = 1:10,
#' dims = 10,
#' top = 500,
#' score = TRUE,
#' show_cols = TRUE,
#' type = "ggplot")
run_APL <- function(obj,
group,
caobj = NULL,
dims = NULL,
nrow = 10,
top = 5000,
score = TRUE,
mark_rows = NULL,
mark_cols = NULL,
reps = 3,
python = FALSE,
row_labs = TRUE,
col_labs = TRUE,
type = "plotly",
show_cols = FALSE,
show_rows = TRUE,
score_cutoff = 0,
score_color = "rainbow",
pd_method = "elbow_rule",
pd_reps = 1,
pd_use = TRUE) {
if (is.null(caobj)) {
caobj <- cacomp(
obj = obj,
coords = TRUE,
top = top,
princ_coords = 3,
dims = dims,
python = python
)
if (is.null(dims) & isTRUE(pd_use)) {
stopifnot("Cannot use scree plot to pick dimensions" = pd_method != "scree_plot")
pd <- pick_dims(
obj = caobj,
mat = obj,
method = pd_method,
reps = pd_reps,
python = python,
return_plot = FALSE
)
message(
"\n Using ",
pd,
" dimensions. Subsetting."
)
dims <- pd
caobj <- subset_dims(caobj, dims)
} else if (is.null(dims) & !isTRUE(pd_use)) {
dims <- caobj@dims
message(
"\n Using ",
dims,
" dimensions. No subsetting performed."
)
} else if (!is.null(dims) & !isTRUE(pd_use)) {
message("\n Using ", dims, " dimensions.")
} else if (!is.null(dims) & isTRUE(pd_use)) {
message("\nBoth dims and pd_use set. Using dimensions specified by dims.")
message("\nUsing ", dims, " dimensions.")
}
} else {
if (!is.empty(caobj@dims) && is.null(dims)) {
if (isTRUE(pd_use)) {
stopifnot(
"Cannot use scree plot to pick dimensions" =
pd_method != "scree_plot"
)
pd <- pick_dims(
obj = caobj,
mat = obj,
method = pd_method,
reps = pd_reps,
python = python,
return_plot = FALSE
)
message(
"\n Using ",
pd,
" dimensions"
)
dims <- pd
caobj <- subset_dims(caobj, dims)
} else {
dims <- caobj@dims
message(
"\n Using ",
dims,
" dimensions. No subsetting performed."
)
}
} else if (!is.empty(caobj@dims) && !is.null(dims)) {
if (dims < caobj@dims) {
if (!isTRUE(pd_use)) {
message("\n Using ", dims, " dimensions.")
} else if (isTRUE(pd_use)) {
message("\nBoth dims and pd_use set. Using dimensions specified by dims.")
message(
"\nUsing ",
dims,
" dimensions."
)
}
caobj <- subset_dims(caobj, dims = dims)
} else if (dims > length(caobj@D)) {
warning(
"dims is larger than the number of available dimensions. ",
"Argument ignored"
)
}
}
if (is.empty(caobj@prin_coords_rows) && !is.empty(caobj@std_coords_rows)) {
caobj <- ca_coords(
caobj = caobj,
dims = dims,
princ_only = TRUE,
princ_coords = 1
)
} else if (is.empty(caobj@prin_coords_rows) ||
is.empty(caobj@std_coords_cols)) {
caobj <- ca_coords(
caobj = caobj,
dims = dims,
princ_only = FALSE,
princ_coords = 1
)
}
}
if (is.empty(caobj@apl_rows) || is.empty(caobj@apl_cols)) {
caobj <- apl_coords(caobj = caobj, group = group)
}
if (is.null(mark_rows)) {
if (score == TRUE) {
caobj <- apl_score(
caobj = caobj,
mat = obj,
dims = caobj@dims,
group = caobj@group,
reps = reps,
python = python
)
mark_rows <- head(caobj@APL_score$Row_num, nrow)
if (is(mark_cols, "character")) {
mark_cols <- match(mark_cols, rownames(caobj@apl_cols))
mark_cols <- na.omit(mark_cols)
if (anyNA(mark_cols)) {
warning(
"Not all names in 'mark_cols' are contained in the row names.",
" Maybe they were filtered out?",
"\nNon-matching values were ignored."
)
}
}
}
} else {
if (is(mark_rows, "character")) {
mark_rows <- match(mark_rows, rownames(caobj@apl_rows))
mark_rows <- na.omit(mark_rows)
if (anyNA(mark_rows)) {
warning(
"Not all names in 'mark_rows' are contained in the row names.",
" Maybe they were filtered out?",
"\n Non-matching values were ignored."
)
}
}
if (is(mark_cols, "character")) {
mark_cols <- match(mark_cols, rownames(caobj@apl_cols))
mark_cols <- na.omit(mark_cols)
if (anyNA(mark_cols)) {
warning(
"Not all names in 'mark_cols' are contained in the row names.",
" Maybe they were filtered out?",
"\n Non-matching values were ignored."
)
}
}
}
if (isTRUE(col_labs) & is.empty(mark_cols)) {
mark_cols <- caobj@group
}
p <- apl(
caobj = caobj,
type = type,
rows_idx = mark_rows,
cols_idx = mark_cols,
row_labs = row_labs,
col_labs = col_labs,
show_score = score,
show_cols = show_cols,
show_rows = show_rows,
score_cutoff = score_cutoff,
score_color = score_color
)
return(p)
}
#' Compute and plot Association Plot
#'
#' @description
#' Computes singular value decomposition and coordinates for
#' the Association Plot.
#'
#' @details
#' The function is a wrapper that calls `cacomp()`, `apl_coords()`,
#' `apl_score()` and finally `apl()` for ease of use.
#' The chosen defaults are most useful for genomics experiments, but for more
#' fine grained control the functions
#' can be also run individually for the same results.
#' If score = FALSE, nrow and reps are ignored. If mark_rows is not NULL score
#' is treated as if FALSE.
#' @return
#' Association Plot (plotly object).
#'
#' @references
#' Association Plots: Visualizing associations in high-dimensional
#' correspondence analysis biplots \cr
#' Elzbieta Gralinska, Martin Vingron \cr
#' bioRxiv 2020.10.23.352096; doi: https://doi.org/10.1101/2020.10.23.352096 \cr
#'
#' @param obj A numeric matrix, Seurat or SingleCellExperiment object. For
#' sequencing a count matrix, gene expression values with genes in rows and
#' samples/cells in columns.
#' Should contain row and column names.
#' @param caobj A "cacomp" object as outputted from `cacomp()`. If not supplied
#' will be calculated. Default NULL.
#' @param dims Integer. Number of dimensions to keep. Default NULL (keeps all
#' dimensions).
#' @param group Numeric/Character. Vector of indices or column names of the
#' columns to calculate centroid/x-axis direction.
#' @param nrow Integer. The top nrow scored row labels will be added to the
#' plot if score = TRUE. Default 10.
#' @param top Integer. Number of most variable rows to retain. Default 5000
#' rows (set NULL to keep all).
#' @param score Logical. Whether rows should be scored and ranked. Ignored when
#' a vector is supplied to mark_rows. Default TRUE.
#' @param mark_rows Character vector. Names of rows that should be highlighted
#' in the plot. If not NULL, score is ignored. Default NULL.
#' @param mark_cols Character vector. Names of cols that should be highlighted
#' in the plot.
#' @param reps Integer. Number of permutations during scoring. Default 3.
#' @param python A logical value indicating whether to use singular value
#' decomposition from the python package torch.
#' This implementation dramatically speeds up computation compared to `svd()`
#' in R.
#' @param row_labs Logical. Whether labels for rows indicated by rows_idx
#' should be labeled with text. Default TRUE.
#' @param col_labs Logical. Whether labels for columns indicated by cols_idx
#' should be labeled with text. Default TRUE.
#' @param type "ggplot"/"plotly". For a static plot a string "ggplot", for an
#' interactive plot "plotly". Default "plotly".
#' @param show_cols Logical. Whether column points should be plotted.
#' @param show_rows Logical. Whether row points should be plotted.
#' @param score_cutoff Numeric. Rows (genes) with a score >= score_cutoff
#' will be colored according to their score if show_score = TRUE.
#' @param score_color Either "rainbow" or "viridis".
#' @param pd_method Which method to use for pick_dims (\link[APL]{pick_dims}).
#' @param pd_reps Number of repetitions performed when using "elbow_rule" in
#' `pick_dims`.
#' (\link[APL]{pick_dims})
#' @param pd_use Whether to use `pick_dims` (\link[APL]{pick_dims}) to determine
#' the number of dimensions. Ignored when `dims` is set by the user.
#' @param ... Arguments forwarded to methods.
#' @export
setGeneric("runAPL", function(obj,
group,
caobj = NULL,
dims = NULL,
nrow = 10,
top = 5000,
score = TRUE, mark_rows = NULL,
mark_cols = caobj@group,
reps = 3,
python = FALSE,
row_labs = TRUE,
col_labs = TRUE,
type = "plotly",
show_cols = FALSE,
show_rows = TRUE,
score_cutoff = 0,
score_color = "rainbow",
pd_method = "elbow_rule",
pd_reps = 1,
pd_use = TRUE,
...) {
standardGeneric("runAPL")
})
#' @export
#' @rdname runAPL
#' @examples
#' set.seed(1234)
#'
#' # Simulate counts
#' cnts <- mapply(function(x){rpois(n = 500, lambda = x)},
#' x = sample(1:100, 50, replace = TRUE))
#' rownames(cnts) <- paste0("gene_", 1:nrow(cnts))
#' colnames(cnts) <- paste0("cell_", 1:ncol(cnts))
#'
#' # (nonsensical) APL
#' runAPL(obj = cnts,
#' group = 1:10,
#' dims = 10,
#' top = 500,
#' score = TRUE,
#' show_cols = TRUE,
#' type = "ggplot")
setMethod(f = "runAPL",
signature=(obj="matrix"),
function(obj,
group,
caobj = NULL,
dims = NULL,
nrow = 10,
top = 5000,
score = TRUE,
mark_rows = NULL,
mark_cols = NULL,
reps = 3,
python = FALSE,
row_labs = TRUE,
col_labs = TRUE,
type = "plotly",
show_cols = FALSE,
show_rows = TRUE,
score_cutoff = 0,
score_color = "rainbow",
pd_method = "elbow_rule",
pd_reps = 1,
pd_use = TRUE,
...) {
if (!is(obj, "matrix") & !is(obj, "dgCMatrix")) {
obj <- as.matrix(obj)
}
stopifnot(is(obj, "matrix") | is(obj, "dgCMatrix"))
run_APL(obj = obj,
caobj = caobj,
dims = dims,
group = group,
mark_rows = mark_rows,
mark_cols = mark_cols,
nrow = nrow,
top = top,
score = score,
reps = reps,
python = python,
row_labs = row_labs,
col_labs = col_labs,
type = type,
show_cols = show_cols,
show_rows = show_rows,
score_cutoff = score_cutoff,
score_color = score_color,
pd_method = pd_method,
pd_reps = pd_reps,
pd_use = pd_use)
})
#' @description
#' runAPL.SingleCellExperiment: Computes singular value decomposition and
#' coordinates for the Association Plot from SingleCellExperiment objects with
#' reducedDim(obj, "CA") slot (optional).
#'
#' @param assay Character. The assay from which extract the count matrix for
#' SVD, e.g. "RNA" for Seurat objects or "counts"/"logcounts" for
#' SingleCellExperiments.
#'
#' @rdname runAPL
#' @export
#' @examples
#'
#' ########################
#' # SingleCellExperiment #
#' ########################
#' library(SingleCellExperiment)
#' set.seed(1234)
#'
#' # Simulate counts
#' cnts <- mapply(function(x){rpois(n = 500, lambda = x)},
#' x = sample(1:100, 50, replace = TRUE))
#' rownames(cnts) <- paste0("gene_", 1:nrow(cnts))
#' colnames(cnts) <- paste0("cell_", 1:ncol(cnts))
#'
#' sce <- SingleCellExperiment(assays=list(counts=cnts))
#'
#' # (nonsensical) APL
#' runAPL(obj = sce,
#' group = 1:10,
#' dims = 10,
#' top = 500,
#' score = TRUE,
#' show_cols = TRUE,
#' type = "ggplot",
#' assay = "counts")
setMethod(f = "runAPL",
signature=(obj="SingleCellExperiment"),
function(obj,
group,
caobj = NULL,
dims = NULL,
nrow = 10,
top = 5000,
score = TRUE,
mark_rows = NULL,
mark_cols = NULL,
reps = 3,
python = FALSE,
row_labs = TRUE,
col_labs = TRUE,
type = "plotly",
show_cols = FALSE,
show_rows = TRUE,
score_cutoff = 0,
score_color = "rainbow",
pd_method = "elbow_rule",
pd_reps = 1,
pd_use = TRUE,
...,
assay = "counts"){
stopifnot("obj doesn't belong to class 'SingleCellExperiment'" =
is(obj, "SingleCellExperiment"))
mat <- SummarizedExperiment::assay(obj, assay)
if ("CA" %in% SingleCellExperiment::reducedDimNames(obj)){
caobj <- as.cacomp(obj, assay = assay)
}
if (!is(mat, "matrix") & !is(mat, "dgCMatrix")) {
mat <- as.matrix(mat)
}
run_APL(obj = mat,
caobj = caobj,
dims = dims,
group = group,
mark_rows = mark_rows,
mark_cols = mark_cols,
nrow = nrow,
top = top,
score = score,
reps = reps,
python = python,
row_labs = row_labs,
col_labs = col_labs,
type = type,
show_cols = show_cols,
show_rows = show_rows,
score_cutoff = score_cutoff,
score_color = score_color,
pd_method = pd_method,
pd_reps = pd_reps,
pd_use = pd_use)
})
#' @description
#' runAPL.Seurat: Computes singular value decomposition and coordinates for
#' the Association Plot from Seurat objects, optionally with a DimReduc Object
#' in the "CA" slot.
#'
#' @param assay Character. The assay from which extract the count matrix for
#' SVD, e.g. "RNA" for Seurat objects or "counts"/"logcounts" for
#' SingleCellExperiments.
#' @param slot character. The Seurat assay slot from which to extract the
#' count matrix.
#' @rdname runAPL
#' @export
#' @examples
#'
#' ###########
#' # Seurat #
#' ###########
#' library(Seurat)
#' set.seed(1234)
#'
#' # Simulate counts
#' cnts <- mapply(function(x){rpois(n = 500, lambda = x)},
#' x = sample(1:100, 50, replace = TRUE))
#' rownames(cnts) <- paste0("gene_", 1:nrow(cnts))
#' colnames(cnts) <- paste0("cell_", 1:ncol(cnts))
#'
#' seu <- CreateSeuratObject(counts = cnts)
#'
#' # (nonsensical) APL
#' runAPL(obj = seu,
#' group = 1:10,
#' dims = 10,
#' top = 500,
#' score = TRUE,
#' show_cols = TRUE,
#' type = "ggplot",
#' assay = "RNA",
#' slot = "counts")
setMethod(f = "runAPL",
signature=(obj="Seurat"),
function(obj,
group,
caobj = NULL,
dims = NULL,
nrow = 10,
top = 5000,
score = TRUE,
mark_rows = NULL,
mark_cols = NULL,
reps = 3,
python = FALSE,
row_labs = TRUE,
col_labs = TRUE,
type = "plotly",
show_cols = FALSE,
show_rows = TRUE,
score_cutoff = 0,
score_color = "rainbow",
pd_method = "elbow_rule",
pd_reps = 1,
pd_use = TRUE,
...,
assay = Seurat::DefaultAssay(obj),
slot = "counts"){
stopifnot("obj doesn't belong to class 'Seurat'" = is(obj, "Seurat"))
seu <- Seurat::GetAssayData(object = obj, assay = assay, slot = slot)
seu <- as.matrix(seu)
if ("CA" %in% Seurat::Reductions(obj)){
caobj <- as.cacomp(obj, assay = assay)
} else {
caobj <- NULL
}
run_APL(obj = seu,
caobj = caobj,
top = top,
dims= dims,
group = group,
score = score,
reps = reps,
python = python,
mark_rows = mark_rows,
mark_cols = mark_cols,
nrow = nrow,
row_labs = row_labs,
col_labs = col_labs,
type = type,
show_cols = show_cols,
show_rows = show_rows,
score_cutoff = score_cutoff,
score_color = score_color,
pd_method = pd_method,
pd_reps = pd_reps,
pd_use = pd_use)
})
#' @export
#' @rdname runAPL
#' @examples
#' set.seed(1234)
#'
#' # Simulate counts
#' cnts <- mapply(function(x){rpois(n = 500, lambda = x)},
#' x = sample(seq(0.01,0.1,by=0.01), 50, replace = TRUE))
#' rownames(cnts) <- paste0("gene_", 1:nrow(cnts))
#' colnames(cnts) <- paste0("cell_", 1:ncol(cnts))
#' cnts <- Matrix::Matrix(cnts)
#'
#' # (nonsensical) APL
#' runAPL(obj = cnts,
#' group = 1:10,
#' dims = 10,
#' top = 500,
#' score = TRUE,
#' show_cols = TRUE,
#' type = "ggplot")
setMethod(f = "runAPL",
signature=(obj="dgCMatrix"),
function(obj,
group,
caobj = NULL,
dims = NULL,
nrow = 10,
top = 5000,
score = TRUE,
mark_rows = NULL,
mark_cols = NULL,
reps = 3,
python = FALSE,
row_labs = TRUE,
col_labs = TRUE,
type = "plotly",
show_cols = FALSE,
show_rows = TRUE,
score_cutoff = 0,
score_color = "rainbow",
pd_method = "elbow_rule",
pd_reps = 1,
pd_use = TRUE,
...) {
stopifnot(is(obj, "dgCMatrix"))
run_APL(obj = obj,
caobj = caobj,
dims = dims,
group = group,
mark_rows = mark_rows,
mark_cols = mark_cols,
nrow = nrow,
top = top,
score = score,
reps = reps,
python = python,
row_labs = row_labs,
col_labs = col_labs,
type = type,
show_cols = show_cols,
show_rows = show_rows,
score_cutoff = score_cutoff,
score_color = score_color,
pd_method = pd_method,
pd_reps = pd_reps,
pd_use = pd_use)
})
ClemensKohl/APL documentation built on May 25, 2024, 3:15 p.m.
R Package Documentation
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Defines functions run_APL apl apl_plotly apl_ggplot apl_topGO apl_score apl_coords
Documented in apl apl_coords apl_ggplot apl_plotly apl_score apl_topGO run_APL
#' @include constructor.R
NULL
#' Calculate Association Plot coordinates
#'
#' @description
#' Calculates the Association Plot coordinates for either the rows,
#' columns or both (default).
#'
#' @details
#' Coordinates (x,y) of row vector \eqn{\vec{r}} are defined as
#' \deqn{x(\vec{r}) := \left|\vec{r}\right|\cos(\phi(\vec{r}))}
#' \deqn{y(\vec{r}) := \left|\vec{r}\right|\sin(\phi(\vec{r}))}
#' The x-direction is determined by calculating the centroid of the columns
#' selected with the indices in "group".
#'
#' @return
#' Returns input "cacomp" object and adds components "apl_rows" and/or
#' "apl_cols" for row and column coordinates.
#' In "group" the indices of the columns used to calculate the
#' centroid are saved.
#'
#' @references
#' Association Plots: Visualizing associations in high-dimensional
#' correspondence analysis biplots
#' Elzbieta Gralinska, Martin Vingron
#' bioRxiv 2020.10.23.352096; doi: https://doi.org/10.1101/2020.10.23.352096
#'
#' @param caobj A "cacomp" object with principal row coordinates and
#' standardized column coordinates calculated.
#' @param group Numeric/Character. Vector of indices or column names of
#' the columns to calculate centroid/x-axis direction.
#' @param calc_rows TRUE/FALSE. Whether apl row coordinates should
#' be calculated. Default TRUE.
#' @param calc_cols TRUE/FALSE. Whether apl column coordinates should
#' be calculated. Default TRUE.
#' @export
#'
#' @examples
#' set.seed(1234)
#' # Simulate scRNAseq data
#' cnts <- data.frame(cell_1 = rpois(10, 5),
#' cell_2 = rpois(10, 10),
#' cell_3 = rpois(10, 20),
#' cell_4 = rpois(10, 20))
#' rownames(cnts) <- paste0("gene_", 1:10)
#' cnts <- as.matrix(cnts)
#'
#' # Run correspondence analysis
#' ca <- cacomp(obj = cnts, princ_coords = 3)
#' # Calculate APL coordinates
#' ca <- apl_coords(ca, group = 3:4)
apl_coords <- function(caobj, group, calc_rows = TRUE, calc_cols = TRUE) {
stopifnot(is(caobj, "cacomp"))
rows <- caobj@prin_coords_rows
cols <- caobj@std_coords_cols
cent <- cols
if (is(group, "numeric")){
subgroup <- cent[group,]
} else if (is(group, "character")){
idx <- match(group, rownames(cent))
idx <- na.omit(idx)
group <- idx
subgroup <- cent[idx, ]
if (anyNA(idx)){
warning("Not all names in 'group' are contained in the column names. ",
"Non-matching values were ignored.")
}
} else {
stop("Parameter group has to be either of type 'numeric' or 'character'.")
}
if (length(group) == 1){
avg_group_coords <- subgroup # single sample
} else {
avg_group_coords <- colMeans(subgroup) # centroid vector.
}
length_vector_group <- sqrt(drop(avg_group_coords %*% avg_group_coords))
length_vector_rows <- sqrt(Matrix::rowSums(rows^2))
length_vector_cols <- sqrt(Matrix::rowSums(cols^2))
if (calc_rows == TRUE){
# message("Calculating APL row coordinates ...")
# r⋅X = |r|*|X|*cosθ
# x(r) = (r⋅X)/|X| = |r|*cosθ
rowx <- drop(rows %*% avg_group_coords)/length_vector_group
# pythagoras, y(r)=b²=c²-a²
rowy <- sqrt(length_vector_rows^2 - rowx^2)
rowx[is.na(rowx)] <- 0
rowy[is.na(rowy)] <- 0
# rowx[is.infinite(rowx)] <- 0
# rowy[is.infinite(rowy)] <- 0
caobj@apl_rows <- cbind("x"=rowx, "y"=rowy)
}
if (calc_cols == TRUE){
# message("Calculating APL column coordinates ...")
colx <- drop(cols %*% avg_group_coords)/length_vector_group
coly <- sqrt(length_vector_cols^2 - colx^2)
colx[is.na(colx)] <- 0
coly[is.na(coly)] <- 0
# colx[is.infinite(colx)] <- 0
# coly[is.infinite(coly)] <- 0
caobj@apl_cols <- cbind("x"=colx, "y"=coly)
}
if (is(group, "numeric")){
caobj@group <- group
} else if (is(group, "character")){
idx <- match(group, rownames(cols))
idx <- na.omit(idx)
caobj@group <- idx
}
stopifnot(validObject(caobj))
return(caobj)
}
#' Find rows most highly associated with a condition
#'
#' @description
#' Ranks rows by a calculated score which balances the association of the row
#' with the condition and how associated it is with other conditions.
#'
#' @details
#' The score is calculated by permuting the values of each row to determine the
#' cutoff angle of the 99% quantile.
#' \deqn{S_{alpha}(x,y)=x-\frac{y}{\tan\alpha}}
#' By default the permutation is repeated 10 times, but for very large matrices
#' this can be reduced.
#' If store_perm is TRUE the permuted data is stored in the cacomp object and
#' can be used for future scoring.
#' @return
#' Returns the input "cacomp" object with "APL_score" component added.
#' APL_score contains a data frame with ranked rows, their score and their
#' original row number.
#'
#' @references
#' Association Plots: Visualizing associations in high-dimensional
#' correspondence analysis biplots \cr
#' Elzbieta Gralinska, Martin Vingron \cr
#' bioRxiv 2020.10.23.352096; doi: https://doi.org/10.1101/2020.10.23.352096
#'
#' @param caobj A "cacomp" object with principal row coordinates and
#' standardized column coordinates calculated.
#' @param mat A numeric matrix. For sequencing a count matrix, gene expression
#' values with genes in rows and samples/cells in columns.
#' Should contain row and column names.
#' @param dims Integer. Number of CA dimensions to retain. Needs to be the same
#' as in caobj!
#' @param group Vector of indices of the columns to calculate centroid/x-axis
#' direction.
#' @param reps Integer. Number of permutations to perform. Default = 10.
#' @param quant Numeric. Single number between 0 and 1 indicating the quantile
#' used to calculate the cutoff. Default 0.99.
#' @param python A logical value indicating whether to use singular-value
#' decomposition from the python package torch.
#' @param store_perm Logical. Whether permuted data should be stored in the CA
#' object.
#' This implementation dramatically speeds up computation compared to `svd()`
#' in R.
#' @export
#'
#' @examples
#' set.seed(1234)
#'
#' # Simulate counts
#' cnts <- mapply(function(x){rpois(n = 500, lambda = x)},
#' x = sample(1:20, 50, replace = TRUE))
#' rownames(cnts) <- paste0("gene_", 1:nrow(cnts))
#' colnames(cnts) <- paste0("cell_", 1:ncol(cnts))
#'
#' # Run correspondence analysis.
#' ca <- cacomp(obj = cnts, princ_coords = 3)
#'
#' # Calculate APL coordinates:
#' ca <- apl_coords(ca, group = 1:10)
#'
#' # Rank genes by S-alpha score
#' ca <- apl_score(ca, mat = cnts)
apl_score <- function(caobj,
mat,
dims = caobj@dims,
group = caobj@group,
reps=10,
quant = 0.99,
python = FALSE,
store_perm = TRUE){
if (!is(caobj,"cacomp")){
stop("Not a CA object. Please run cacomp() and apl_coords() first!")
}
if (is.empty(caobj@apl_rows)){
stop("Please run apl_coords() first!")
}
names <- colnames(mat)
row_num <- nrow(caobj@apl_rows)
margin <- 1
pc <- 1
cc <- FALSE
cr <- TRUE
if (is(group, "character")){
idx <- match(group, names)
idx <- na.omit(idx)
group <- idx
}
if (caobj@dims == 1 && !is.empty(caobj@dims)){
row_num <- 1
}
apl_perm <- data.frame("x" = rep(0, row_num*reps),
"y" = rep(0, row_num*reps))
saved_ca <- list()
pb <- txtProgressBar(min = 0, max = reps, style = 3)
for (k in seq(reps)){
#permute rows and rerun cacomp
if(isTRUE(store_perm) & identical(reps, attr(caobj@permuted_data,'reps'))){
calist <- caobj@permuted_data[[k]][seq_len(3)]
mat <- caobj@permuted_data[[k]]$mat
mat <- mat[rownames(mat) %in% rownames(calist$std_coords_rows),]
caobjp <- recompute(calist, mat)
} else {
mat_perm <- t(apply(mat, margin, FUN=sample))
colnames(mat_perm) <- colnames(mat)
suppressWarnings(caobjp <- cacomp(obj = mat_perm,
python = python,
coords = TRUE,
princ_coords = pc,
dims = dims,
top = caobj@top_rows,
inertia = FALSE))
if(isTRUE(store_perm)){
x <- list("std_coords_cols" = caobjp@std_coords_cols,
"std_coords_rows" = caobjp@std_coords_rows,
"D" = caobjp@D,
"mat" = mat_perm)
saved_ca[[k]] <- x
}
}
caobjp <- apl_coords(caobj = caobjp,
group = group,
calc_cols = cc,
calc_rows = cr)
idx <- ((seq_len(row_num)+((k-1)*row_num)))
apl_perm[idx,] <- caobjp@apl_rows
setTxtProgressBar(pb, k)
}
close(pb)
apl_perm[,3] <- apl_perm[,1]/apl_perm[,2] # cotan between row and x axis
apl_perm[,3][is.na(apl_perm[,3])] <- 0
cutoff_cotan <- quantile(apl_perm[,3], quant)
score <- caobj@apl_rows[,1] - (caobj@apl_rows[,2] * cutoff_cotan)
ranking <- data.frame("Rowname" = rownames(caobj@apl_rows),
"Score" = score,
"Row_num" = seq_len(nrow(caobj@apl_rows)))
ranking <- ranking[order(ranking$Score, decreasing = TRUE),]
ranking$Rank <- seq_len(nrow(ranking))
caobj@APL_score <- ranking
if(isTRUE(store_perm) & !identical(reps, attr(caobj@permuted_data,'reps'))){
caobj@permuted_data <- saved_ca
attr(caobj@permuted_data,'cutoff') <- cutoff_cotan
attr(caobj@permuted_data,'reps') <- reps
}
stopifnot(validObject(caobj))
return(caobj)
}
#' Run Gene overrepresentation analysis with topGO
#'
#' @description
#' This function uses the Kolmogorov-Smirnov test as implemented by the package
#' topGO to test for overrepresentation in Gene Ontology gene sets.
#'
#' @details
#' For a chosen group of cells/samples,
#' the top 'ngenes' group specific genes are used for gene overrepresentation
#' analysis.
#' The genes are ranked either by the precomputed APL score, or, if
#' not available by their APL x-coordinates.
#'
#' @return
#' A data.frame containing the gene sets with the highest overrepresentation.
#'
#' @references
#' Adrian Alexa and Jorg Rahnenfuhrer \cr
#' topGO: Enrichment Analysis for Gene Ontology. \cr
#' R package version 2.42.0.
#'
#' @param caobj A "cacomp" object with principal row coordinates and
#' standardized column coordinates calculated.
#' @param ontology Character string. Chooses GO sets for 'BP'
#' (biological processes), 'CC' (cell compartment) or 'MF' (molecular function).
#' @param organism Character string. Either 'hs' (homo sapiens), 'mm'
#' (mus musculus) or the name of the organism package such as 'org.*.eg.db'.
#' @param ngenes Numeric. Number of top ranked genes to test for
#' overrepresentation.
#' @param score_cutoff numeric. S-alpha score cutoff. Only genes with a score
#' larger will be tested.
#' @param use_coords Logical. Whether the x-coordinates of the row APL
#' coordinates should be used for ranking.
#' Only recommended when no S-alpha score (see apl_score()) can be calculated.
#' @param return_plot Logical. Whether a plot of significant gene sets should
#' be additionally returned.
#' @param top_res Numeric. Number of top scoring genes to plot.
#'
#' @export
#' @examples
#' library(Seurat)
#' set.seed(1234)
#' cnts <- GetAssayData(pbmc_small, assay = "RNA", slot = "counts")
#' cnts <- as.matrix(cnts)
#'
#' # Run CA on example from Seurat
#'
#' ca <- cacomp(pbmc_small,
#' princ_coords = 3,
#' return_input = FALSE,
#' assay = "RNA",
#' slot = "counts")
#'
#' grp <- which(Idents(pbmc_small) == 2)
#' ca <- apl_coords(ca, group = grp)
#' ca <- apl_score(ca,
#' mat = cnts)
#'
#' enr <- apl_topGO(ca,
#' ontology = "BP",
#' organism = "hs")
#'
#' plot_enrichment(enr)
apl_topGO <- function(caobj,
ontology,
organism="hs",
ngenes = 1000,
score_cutoff = 0,
use_coords = FALSE,
return_plot = FALSE,
top_res = 15){
topGO::groupGOTerms()
if(!is.empty(caobj@APL_score) & !isTRUE(use_coords)){
Score_ord <- caobj@APL_score[order(caobj@APL_score$Score, decreasing=TRUE),]
sel <- which(!Score_ord$Score >= score_cutoff)[1]
sel <- sel-1
ranked_genes <- seq_len(nrow(Score_ord))
names(ranked_genes) <- Score_ord$Rowname
} else if (isTRUE(use_coords) & is.empty(caobj@APL_score)) {
if(is.empty(caobj@apl_rows)){
stop("No APL coordinates found for rows. Please first run apl_coords.\n")
}
if (ngenes > nrow(caobj@apl_rows)){
stop("ngenes is larger than the total number of genes.\n")
} else if (ngenes == nrow(caobj@apl_rows)){
warning("You have selected all available genes.\n")
}
ranked_genes <- seq_len(nrow(caobj@apl_rows))
names(ranked_genes) <- rownames(caobj@apl_rows)[order(caobj@apl_rows[,1],
decreasing = TRUE)]
sel <- ngenes
} else {
stop("APL scores not present but use_coords set to FALSE.\n")
}
if (organism == "hs"){
organism <- "org.Hs.eg.db"
} else if (organism == "mm"){
organism <- "org.Mm.eg.db"
} else {
warning("Custom organism chosen.\n")
organism <- organism
}
if (!ontology %in% c("BP", "CC", "MF")){
stop("Please choose one of the following ontologies: 'BP', 'CC' or 'MF'.\n")
}
gene_sets <- topGO::annFUN.org(whichOnto=ontology,
feasibleGenes=NULL,
mapping=organism,
ID="symbol")
GOdata <- new("topGOdata",
ontology = ontology,
allGenes = ranked_genes,
annot = topGO::annFUN.GO2genes,
GO2genes = gene_sets,
geneSelectionFun = function(x) x<sel,
nodeSize=5)
results_test <- topGO::runTest(GOdata,
algorithm = "elim",
statistic = "fisher")
goEnrichment <- topGO::GenTable(GOdata,
raw.p.value = results_test,
topNodes = length(results_test@score),
numChar = 1200)
if (isTRUE(return_plot)){
if(top_res > length(results_test@score)){
warning("More top nodes selected via top_res than available. ",
"Returning max. number of nodes instead.\n")
top_res <- min(top_res, length(results_test@score))
}
topGO::showSigOfNodes(GOdata,
score(results_test),
firstSigNodes = top_res,
useInfo = 'def')
}
return(goEnrichment)
}
#'
#' Plot Association Plot with ggplot
#' @description
#' Uses ggplot to plot an Association Plot
#'
#' @param rows Row APL-coordinates
#' @param rows_group Row AP-coordinates to highlight
#' @param rows_scored Row AP-coordinates of rows above a score cutoff.
#' @param cols Column AP-coordinates
#' @param cols_group Column AP-coordinates for the group to be highlighted.
#' @param rows_color Color for rows
#' @param rows_high_color Color for rows to be highlighted.
#' @param cols_color Column points color.
#' @param cols_high_color Color for column points to be highlighted..
#' @param score_color Color scheme for row points with a score.
#' @param row_labs Logical. Whether labels for rows indicated by rows_idx
#' should be labeled with text. Default TRUE.
#' @param col_labs Logical. Whether labels for columns indicated by cols_idx
#' shouls be labeled with text. Default FALSE.
#' @param show_score Logical. Whether the S-alpha score should be shown in
#' the plot.
#' @param show_cols Logical. Whether column points should be plotted.
#' @param show_rows Logical. Whether row points should be plotted.
#'
#' @returns
#' ggplot Association Plot
apl_ggplot <- function(rows,
rows_group = NULL,
cols,
cols_group = NULL,
rows_scored = NULL,
rows_color = "#0066FF",
rows_high_color = "#FF0000",
cols_color = "#601A4A",
cols_high_color = "#EE442F",
score_color = "rainbow",
row_labs = FALSE,
col_labs = FALSE,
show_score = FALSE,
show_cols = FALSE,
show_rows = TRUE){
p <- ggplot2::ggplot()
if (isTRUE(show_rows)){
p <- p +
ggplot2::geom_point(data=rows,
ggplot2::aes(x=.data$x, y=.data$y),
color = rows_color,
alpha = 0.7,
shape = 1) #16 point, 1 circle.
if (isTRUE(show_score)){
p <- p +
ggplot2::geom_point(data=rows_scored,
ggplot2::aes(x=.data$x,
y=.data$y,
fill = .data$Score),
alpha = 0.7,
shape = 21,
stroke = 0.5)
if (score_color == "rainbow"){
hex <- c("#00007F",
"blue",
"#007FFF",
"cyan",
"#7FFF7F",
"yellow",
"#FF7F00",
"red",
"#7F0000")
p <- p +
ggplot2::scale_fill_gradientn(colours = hex,
name = expression(S*alpha))
} else if (score_color == "viridis"){
p <- p +
ggplot2::scale_color_viridis_c(option = "D")
}
}
if (!is.null(rows_group)){
p <- p +
ggplot2::geom_point(data=rows_group,
ggplot2::aes(x=.data$x, y=.data$y),
color = rows_high_color,
shape = 19)
if(isTRUE(row_labs)){
p <- p +
ggrepel::geom_text_repel(data = rows_group,
ggplot2::aes(x=.data$x,
y=.data$y,
label=.data$rownms),
color = rows_high_color,
max.overlaps = Inf)
}
}
}
if (isTRUE(show_cols)){
p <- p +
ggplot2::geom_point(data=cols,
ggplot2::aes(x=.data$x, y=.data$y),
color = cols_color,
shape = 4)
if (!is.null(cols_group)){
p <- p +
ggplot2::geom_point(data=cols_group,
ggplot2::aes(x=.data$x, y=.data$y),
color = cols_high_color,
shape = 4)
if(col_labs == TRUE){
p <- p +
ggrepel::geom_text_repel(data=cols_group,
ggplot2::aes(x=.data$x,
y=.data$y,
label=.data$rownms),
color = cols_high_color)
}
}
}
p <- p +
ggplot2::labs(title="Association Plot") +
ggplot2::theme_bw()
return(p)
}
#'
#' Plot Association Plot with plotly
#' @description
#' Uses plotly to generate an interactive Association Plot
#'
#' @param rows Row APL-coordinates
#' @param rows_group Row AP-coordinates to highlight
#' @param rows_scored Row AP-coordinates of rows above a score cutoff.
#' @param cols Column AP-coordinates
#' @param cols_group Column AP-coordinates for the group to be highlighted.
#' @param rows_color Color for rows
#' @param rows_high_color Color for rows to be highlighted.
#' @param cols_color Column points color.
#' @param cols_high_color Color for column points to be highlighted.
#' @param score_color Color scheme for row points with a score.
#' @param row_labs Logical. Whether labels for rows indicated by rows_idx
#' should be labeled with text. Default TRUE.
#' @param col_labs Logical. Whether labels for columns indicated by cols_idx
#' shouls be labeled with text. Default FALSE.
#' @param show_score Logical. Whether the S-alpha score should be shown in
#' the plot.
#' @param show_cols Logical. Whether column points should be plotted.
#' @param show_rows Logical. Whether row points should be plotted.
#'
#' @returns
#' Interactive plotly Association Plot
apl_plotly <- function(rows,
rows_group =NULL,
cols,
cols_group,
rows_scored = NULL,
rows_color = "#0066FF",
rows_high_color = "#FF0000",
cols_color = "#601A4A",
cols_high_color = "#EE442F",
score_color = "rainbow",
row_labs = FALSE,
col_labs = FALSE,
show_score = FALSE,
show_cols = FALSE,
show_rows = TRUE){
if (row_labs == FALSE){
rlabs <- 'markers'
rowfont <- NULL
} else {
rlabs <- 'markers+text'
rowfont <- list(color=rows_high_color)
}
if (col_labs == FALSE){
clabs <- 'markers'
colfont <- NULL
} else {
clabs <- 'markers+text'
colfont <- list(color = cols_high_color)
}
p <- plotly::plot_ly()
if(isTRUE(show_rows)){
color_fix <- rows_color
colors_fun <- NULL
color_bar <- NULL
sym <- "circle-open"
p <- p %>%
plotly::add_trace(data = rows,
x = ~x,
y = ~y,
mode = 'markers',
text = rows$rownms,
opacity = 0.7,
textposition = "left",
marker = list(color = color_fix,
colorscale = colors_fun,
symbol = sym,
colorbar = color_bar,
size = 5),
name = 'genes',
hoverinfo = 'text',
type = 'scatter')
if (isTRUE(show_score)){
if (score_color == "rainbow"){
colors_fun <- "Jet"
} else if (score_color == "viridis"){
colors_fun <- 'Viridis'
}
color_fix <- as.formula("~Score")
color_bar <- list(title = "S\u03B1", len=0.5)
sym <- "circle"
p <- p %>%
plotly::add_trace(data = rows_scored,
x = ~x,
y = ~y,
mode = 'markers',
text = rows_scored$rownms,
opacity = 0.7,
textposition = "left",
marker = list(color = color_fix,
colorscale = colors_fun,
symbol = sym,
colorbar = color_bar,
size = 5,
line = list(color = '#000000', #black
width = 0.5)),
name = 'genes (scored)',
hoverinfo = 'text',
type = 'scatter')
}
if (!is.null(rows_group)){
p <- p %>%
plotly::add_trace(data = rows_group,
x = ~x,
y = ~y,
mode = rlabs,
text = rows_group$rownms,
textposition = "left",
textfont=rowfont,
marker = list(symbol = 'circle',
color = rows_high_color,
size = 5),
name = 'marked genes',
hoverinfo = 'text',
type = 'scatter')
}
}
if(isTRUE(show_cols)){
p <- p %>%
plotly::add_trace(data=cols,
x = ~x,
y = ~y,
mode = 'markers',
text = cols$rownms,
textposition = "left",
marker = list(color = cols_color,
symbol = 'x',
size = 5),
name = 'samples',
hoverinfo = 'text',
type = 'scatter')
if (!is.null(cols_group)){
p <- p %>% plotly::add_trace(data = cols_group,
x = ~x,
y = ~y,
mode = clabs,
text = cols_group$rownms,
textposition = "left",
textfont=colfont,
marker = list(symbol = 'x',
color = cols_high_color,
size = 5),
name = 'marked samples',
hoverinfo = 'text',
type = 'scatter')
}
}
p <- p %>%
plotly::layout(title = paste('Association Plot'),
xaxis = list(title = 'Distance from origin (x)',
rangemode = "tozero"),
yaxis = list(title = 'Distance from gene to sample line (y)',
rangemode = "tozero"),
showlegend = TRUE)
return(p)
}
#' Association Plot
#'
#' @description
#' Plot an Association Plot for the chosen columns.
#'
#' @details
#' For an interactive plot type="plotly" can be chosen, otherwise a static plot
#' will be returned.
#' The row and column coordinates have to be already calculated by
#' `apl_coords()`.
#'
#' @return
#' Either a ggplot or plotly object.
#'
#' @references
#' Association Plots: Visualizing associations in high-dimensional
#' correspondence analysis biplots \cr
#' Elzbieta Gralinska, Martin Vingron \cr
#' bioRxiv 2020.10.23.352096; doi: https://doi.org/10.1101/2020.10.23.352096 \cr
#'
#' @param caobj An object of class "cacomp" and "APL" with apl
#' coordinates calculated.
#' @param type "ggplot"/"plotly". For a static plot a string "ggplot",
#' for an interactive plot "plotly". Default "ggplot".
#' @param rows_idx numeric/character vector.
#' Indices or names of the rows that should be labelled. Default NULL.
#' @param cols_idx numeric/character vector.
#' Indices or names of the columns that should be labelled.
#' Default is only to label columns making up the centroid: caobj@group.
#' @param row_labs Logical. Whether labels for rows indicated by rows_idx
#' should be labeled with text. Default TRUE.
#' @param col_labs Logical. Whether labels for columns indicated by cols_idx
#' shouls be labeled with text. Default FALSE.
#' @param show_score Logical. Whether the S-alpha score should be shown in
#' the plot.
#' @param show_cols Logical. Whether column points should be plotted.
#' @param show_rows Logical. Whether row points should be plotted.
#' @param score_cutoff Numeric. Rows (genes) with a score >= score_cutoff will
#' be colored according to their score if show_score = TRUE.
#' @param score_color Either "rainbow" or "viridis".
#' @export
#' @examples
#' set.seed(1234)
#'
#' # Simulate counts
#' cnts <- mapply(function(x){rpois(n = 500, lambda = x)},
#' x = sample(1:100, 50, replace = TRUE))
#' rownames(cnts) <- paste0("gene_", 1:nrow(cnts))
#' colnames(cnts) <- paste0("cell_", 1:ncol(cnts))
#'
#' # Run correspondence analysis
#' ca <- cacomp(obj = cnts, princ_coords = 3)
#'
#' # Calculate APL coordinates for arbitrary group
#' ca <- apl_coords(ca, group = 1:10)
#'
#' # plot results
#' # Note:
#' # Due to random gene expression & group, no highly
#' # associated genes are visible.
#' apl(ca, type = "ggplot")
apl <- function(caobj,
type="ggplot",
rows_idx = NULL,
cols_idx = caobj@group,
row_labs = FALSE,
col_labs = FALSE,
show_score = FALSE,
show_cols = FALSE,
show_rows = TRUE,
score_cutoff = 0,
score_color = "rainbow"){
if (!is(caobj,"cacomp")){
stop("Not a CA object. Please run cacomp() and apl_coords() first!")
}
if (is.empty(caobj@apl_rows) || is.empty(caobj@apl_cols)){
stop("Please run apl_coords() first!")
}
if (is(rows_idx, "character")){
names <- rownames(caobj@apl_rows)
idx <- match(rows_idx, names)
idx <- na.omit(idx)
rows_idx <- idx
}
if (is(cols_idx, "character")){
names <- rownames(caobj@apl_cols)
idx <- match(cols_idx, names)
idx <- na.omit(idx)
cols_idx <- idx
}
if (is.numeric(cols_idx)){
group_cols <- data.frame(rownms = rownames(caobj@apl_cols)[cols_idx],
x = caobj@apl_cols[cols_idx,"x"],
y = caobj@apl_cols[cols_idx,"y"],
row.names = NULL)
} else {
group_cols <- NULL
}
if(is.numeric(rows_idx)){
group_rows <- data.frame(rownms = rownames(caobj@apl_rows)[rows_idx],
x = caobj@apl_rows[rows_idx,"x"],
y = caobj@apl_rows[rows_idx,"y"],
row.names = NULL)
} else {
group_rows <- NULL
}
rows_color <- "#0066FF"
rows_high_color <- "#FF0000"
cols_color <- "#601A4A"
cols_high_color <- "#EE442F"
apl_rows.tmp <- data.frame(rownms = rownames(caobj@apl_rows),
caobj@apl_rows)
apl_cols.tmp <- data.frame(rownms = rownames(caobj@apl_cols),
caobj@apl_cols)
if (isTRUE(show_score)) {
apl_scores <- caobj@APL_score$Score[order(caobj@APL_score$Row_num)]
apl_rows.tmp$Score <- apl_scores
idx <- which(apl_scores >= score_cutoff)
if (is.empty(idx)) {
show_score <- FALSE
warning("No rows with a Score >= score_cutoff of ",
score_cutoff,
". Choose lower cutoff.")
} else {
apl_scored.tmp <- apl_rows.tmp[idx,]
apl_rows.tmp <- apl_rows.tmp[-idx,]
}
}
if (type == "ggplot"){
p <- apl_ggplot(rows = apl_rows.tmp,
rows_group = group_rows,
cols = apl_cols.tmp,
cols_group = group_cols,
rows_scored = apl_scored.tmp,
rows_color = rows_color,
rows_high_color = rows_high_color,
cols_color = cols_color,
cols_high_color = cols_high_color,
score_color = score_color,
row_labs = row_labs,
col_labs = col_labs,
show_score = show_score,
show_cols = show_cols,
show_rows = show_rows)
return(p)
} else if (type == "plotly"){
p <- apl_plotly(rows = apl_rows.tmp,
rows_group = group_rows,
cols = apl_cols.tmp,
cols_group = group_cols,
rows_scored = apl_scored.tmp,
rows_color = rows_color,
rows_high_color = rows_high_color,
cols_color = cols_color,
cols_high_color = cols_high_color,
score_color = score_color,
row_labs = row_labs,
col_labs = col_labs,
show_score = show_score,
show_cols = show_cols,
show_rows = show_rows)
return(p)
} else {
stop("Please specify plot = \"ggplot\" or \"plotly\".",
" Other options are not accepted.")
}
}
#' Internal function to compute and plot Association Plot
#'
#' @description
#' Computes singular value decomposition and coordinates for
#' the Association Plot.
#'
#' @details
#' The function is a wrapper that calls `cacomp()`, `apl_coords()`,
#' `apl_score()` and finally `apl()` for ease of use.
#' The chosen defaults are most useful for genomics experiments, but for more
#' fine grained control the functions
#' can be also run individually for the same results.
#' If score = FALSE, nrow and reps are ignored. If mark_rows is not NULL score
#' is treated as if FALSE.
#' @return
#' Association Plot (plotly object).
#'
#' @references
#' Association Plots: Visualizing associations in high-dimensional
#' correspondence analysis biplots \cr
#' Elzbieta Gralinska, Martin Vingron \cr
#' bioRxiv 2020.10.23.352096; doi: https://doi.org/10.1101/2020.10.23.352096 \cr
#'
#' @param obj A numeric matrix. For sequencing usually a count matrix,
#' gene expression values with genes in rows and samples/cells in columns.
#' Should contain row and column names.
#'
#' @param caobj A "cacomp" object as outputted from `cacomp()`. If not supplied
#' will be calculated. Default NULL.
#'
#' @param dims Integer. Number of dimensions to keep. Default NULL (keeps all
#' dimensions).
#'
#' @param group Numeric/Character. Vector of indices or column names of the
#' columns to calculate centroid/x-axis direction.
#'
#' @param nrow Integer. The top nrow scored row labels will be added to the
#' plot if score = TRUE. Default 10.
#'
#' @param top Integer. Number of most variable rows to retain. Default 5000
#' rows (set NULL to keep all).
#'
#' @param score Logical. Whether rows should be scored and ranked. Ignored when
#' a vector is supplied to mark_rows. Default TRUE.
#'
#' @param mark_rows Character vector. Names of rows that should be highlighted
#' in the plot. If not NULL, score is ignored. Default NULL.
#'
#' @param mark_cols Character vector. Names of cols that should be highlighted
#' in the plot.
#'
#' @param reps Integer. Number of permutations during scoring. Default 3.
#'
#' @param python A logical value indicating whether to use singular value
#' decomposition from the python package torch.
#' This implementation dramatically speeds up computation compared to `svd()`
#' in R.
#'
#' @param row_labs Logical. Whether labels for rows indicated by rows_idx
#' should be labeled with text. Default TRUE.
#'
#' @param col_labs Logical. Whether labels for columns indicated by cols_idx
#' should be labeled with text. Default TRUE.
#'
#' @param type "ggplot"/"plotly". For a static plot a string "ggplot", for an
#' interactive plot "plotly". Default "plotly".
#'
#' @param show_cols Logical. Whether column points should be plotted.
#'
#' @param show_rows Logical. Whether row points should be plotted.
#'
#' @param score_cutoff Numeric. Rows (genes) with a score >= score_cutoff
#' will be colored according to their score if show_score = TRUE.
#'
#' @param score_color Either "rainbow" or "viridis".
#'
#' @param pd_method Which method to use for pick_dims (\link[APL]{pick_dims}).
#'
#' @param pd_reps Number of repetitions performed when using "elbow_rule" in
#' `pick_dims`.
#' (\link[APL]{pick_dims})
#'
#' @param pd_use Whether to use `pick_dims` (\link[APL]{pick_dims}) to determine
#' the number of dimensions. Ignored when `dims` is set by the user.
#'
#' @examples
#' set.seed(1234)
#'
#' # Simulate counts
#' cnts <- mapply(function(x){rpois(n = 500, lambda = x)},
#' x = sample(1:100, 50, replace = TRUE))
#' rownames(cnts) <- paste0("gene_", 1:nrow(cnts))
#' colnames(cnts) <- paste0("cell_", 1:ncol(cnts))
#'
#' # (nonsensical) APL
#' APL:::run_APL(obj = cnts,
#' group = 1:10,
#' dims = 10,
#' top = 500,
#' score = TRUE,
#' show_cols = TRUE,
#' type = "ggplot")
run_APL <- function(obj,
group,
caobj = NULL,
dims = NULL,
nrow = 10,
top = 5000,
score = TRUE,
mark_rows = NULL,
mark_cols = NULL,
reps = 3,
python = FALSE,
row_labs = TRUE,
col_labs = TRUE,
type = "plotly",
show_cols = FALSE,
show_rows = TRUE,
score_cutoff = 0,
score_color = "rainbow",
pd_method = "elbow_rule",
pd_reps = 1,
pd_use = TRUE) {
if (is.null(caobj)) {
caobj <- cacomp(
obj = obj,
coords = TRUE,
top = top,
princ_coords = 3,
dims = dims,
python = python
)
if (is.null(dims) & isTRUE(pd_use)) {
stopifnot("Cannot use scree plot to pick dimensions" = pd_method != "scree_plot")
pd <- pick_dims(
obj = caobj,
mat = obj,
method = pd_method,
reps = pd_reps,
python = python,
return_plot = FALSE
)
message(
"\n Using ",
pd,
" dimensions. Subsetting."
)
dims <- pd
caobj <- subset_dims(caobj, dims)
} else if (is.null(dims) & !isTRUE(pd_use)) {
dims <- caobj@dims
message(
"\n Using ",
dims,
" dimensions. No subsetting performed."
)
} else if (!is.null(dims) & !isTRUE(pd_use)) {
message("\n Using ", dims, " dimensions.")
} else if (!is.null(dims) & isTRUE(pd_use)) {
message("\nBoth dims and pd_use set. Using dimensions specified by dims.")
message("\nUsing ", dims, " dimensions.")
}
} else {
if (!is.empty(caobj@dims) && is.null(dims)) {
if (isTRUE(pd_use)) {
stopifnot(
"Cannot use scree plot to pick dimensions" =
pd_method != "scree_plot"
)
pd <- pick_dims(
obj = caobj,
mat = obj,
method = pd_method,
reps = pd_reps,
python = python,
return_plot = FALSE
)
message(
"\n Using ",
pd,
" dimensions"
)
dims <- pd
caobj <- subset_dims(caobj, dims)
} else {
dims <- caobj@dims
message(
"\n Using ",
dims,
" dimensions. No subsetting performed."
)
}
} else if (!is.empty(caobj@dims) && !is.null(dims)) {
if (dims < caobj@dims) {
if (!isTRUE(pd_use)) {
message("\n Using ", dims, " dimensions.")
} else if (isTRUE(pd_use)) {
message("\nBoth dims and pd_use set. Using dimensions specified by dims.")
message(
"\nUsing ",
dims,
" dimensions."
)
}
caobj <- subset_dims(caobj, dims = dims)
} else if (dims > length(caobj@D)) {
warning(
"dims is larger than the number of available dimensions. ",
"Argument ignored"
)
}
}
if (is.empty(caobj@prin_coords_rows) && !is.empty(caobj@std_coords_rows)) {
caobj <- ca_coords(
caobj = caobj,
dims = dims,
princ_only = TRUE,
princ_coords = 1
)
} else if (is.empty(caobj@prin_coords_rows) ||
is.empty(caobj@std_coords_cols)) {
caobj <- ca_coords(
caobj = caobj,
dims = dims,
princ_only = FALSE,
princ_coords = 1
)
}
}
if (is.empty(caobj@apl_rows) || is.empty(caobj@apl_cols)) {
caobj <- apl_coords(caobj = caobj, group = group)
}
if (is.null(mark_rows)) {
if (score == TRUE) {
caobj <- apl_score(
caobj = caobj,
mat = obj,
dims = caobj@dims,
group = caobj@group,
reps = reps,
python = python
)
mark_rows <- head(caobj@APL_score$Row_num, nrow)
if (is(mark_cols, "character")) {
mark_cols <- match(mark_cols, rownames(caobj@apl_cols))
mark_cols <- na.omit(mark_cols)
if (anyNA(mark_cols)) {
warning(
"Not all names in 'mark_cols' are contained in the row names.",
" Maybe they were filtered out?",
"\nNon-matching values were ignored."
)
}
}
}
} else {
if (is(mark_rows, "character")) {
mark_rows <- match(mark_rows, rownames(caobj@apl_rows))
mark_rows <- na.omit(mark_rows)
if (anyNA(mark_rows)) {
warning(
"Not all names in 'mark_rows' are contained in the row names.",
" Maybe they were filtered out?",
"\n Non-matching values were ignored."
)
}
}
if (is(mark_cols, "character")) {
mark_cols <- match(mark_cols, rownames(caobj@apl_cols))
mark_cols <- na.omit(mark_cols)
if (anyNA(mark_cols)) {
warning(
"Not all names in 'mark_cols' are contained in the row names.",
" Maybe they were filtered out?",
"\n Non-matching values were ignored."
)
}
}
}
if (isTRUE(col_labs) & is.empty(mark_cols)) {
mark_cols <- caobj@group
}
p <- apl(
caobj = caobj,
type = type,
rows_idx = mark_rows,
cols_idx = mark_cols,
row_labs = row_labs,
col_labs = col_labs,
show_score = score,
show_cols = show_cols,
show_rows = show_rows,
score_cutoff = score_cutoff,
score_color = score_color
)
return(p)
}
#' Compute and plot Association Plot
#'
#' @description
#' Computes singular value decomposition and coordinates for
#' the Association Plot.
#'
#' @details
#' The function is a wrapper that calls `cacomp()`, `apl_coords()`,
#' `apl_score()` and finally `apl()` for ease of use.
#' The chosen defaults are most useful for genomics experiments, but for more
#' fine grained control the functions
#' can be also run individually for the same results.
#' If score = FALSE, nrow and reps are ignored. If mark_rows is not NULL score
#' is treated as if FALSE.
#' @return
#' Association Plot (plotly object).
#'
#' @references
#' Association Plots: Visualizing associations in high-dimensional
#' correspondence analysis biplots \cr
#' Elzbieta Gralinska, Martin Vingron \cr
#' bioRxiv 2020.10.23.352096; doi: https://doi.org/10.1101/2020.10.23.352096 \cr
#'
#' @param obj A numeric matrix, Seurat or SingleCellExperiment object. For
#' sequencing a count matrix, gene expression values with genes in rows and
#' samples/cells in columns.
#' Should contain row and column names.
#' @param caobj A "cacomp" object as outputted from `cacomp()`. If not supplied
#' will be calculated. Default NULL.
#' @param dims Integer. Number of dimensions to keep. Default NULL (keeps all
#' dimensions).
#' @param group Numeric/Character. Vector of indices or column names of the
#' columns to calculate centroid/x-axis direction.
#' @param nrow Integer. The top nrow scored row labels will be added to the
#' plot if score = TRUE. Default 10.
#' @param top Integer. Number of most variable rows to retain. Default 5000
#' rows (set NULL to keep all).
#' @param score Logical. Whether rows should be scored and ranked. Ignored when
#' a vector is supplied to mark_rows. Default TRUE.
#' @param mark_rows Character vector. Names of rows that should be highlighted
#' in the plot. If not NULL, score is ignored. Default NULL.
#' @param mark_cols Character vector. Names of cols that should be highlighted
#' in the plot.
#' @param reps Integer. Number of permutations during scoring. Default 3.
#' @param python A logical value indicating whether to use singular value
#' decomposition from the python package torch.
#' This implementation dramatically speeds up computation compared to `svd()`
#' in R.
#' @param row_labs Logical. Whether labels for rows indicated by rows_idx
#' should be labeled with text. Default TRUE.
#' @param col_labs Logical. Whether labels for columns indicated by cols_idx
#' should be labeled with text. Default TRUE.
#' @param type "ggplot"/"plotly". For a static plot a string "ggplot", for an
#' interactive plot "plotly". Default "plotly".
#' @param show_cols Logical. Whether column points should be plotted.
#' @param show_rows Logical. Whether row points should be plotted.
#' @param score_cutoff Numeric. Rows (genes) with a score >= score_cutoff
#' will be colored according to their score if show_score = TRUE.
#' @param score_color Either "rainbow" or "viridis".
#' @param pd_method Which method to use for pick_dims (\link[APL]{pick_dims}).
#' @param pd_reps Number of repetitions performed when using "elbow_rule" in
#' `pick_dims`.
#' (\link[APL]{pick_dims})
#' @param pd_use Whether to use `pick_dims` (\link[APL]{pick_dims}) to determine
#' the number of dimensions. Ignored when `dims` is set by the user.
#' @param ... Arguments forwarded to methods.
#' @export
setGeneric("runAPL", function(obj,
group,
caobj = NULL,
dims = NULL,
nrow = 10,
top = 5000,
score = TRUE, mark_rows = NULL,
mark_cols = caobj@group,
reps = 3,
python = FALSE,
row_labs = TRUE,
col_labs = TRUE,
type = "plotly",
show_cols = FALSE,
show_rows = TRUE,
score_cutoff = 0,
score_color = "rainbow",
pd_method = "elbow_rule",
pd_reps = 1,
pd_use = TRUE,
...) {
standardGeneric("runAPL")
})
#' @export
#' @rdname runAPL
#' @examples
#' set.seed(1234)
#'
#' # Simulate counts
#' cnts <- mapply(function(x){rpois(n = 500, lambda = x)},
#' x = sample(1:100, 50, replace = TRUE))
#' rownames(cnts) <- paste0("gene_", 1:nrow(cnts))
#' colnames(cnts) <- paste0("cell_", 1:ncol(cnts))
#'
#' # (nonsensical) APL
#' runAPL(obj = cnts,
#' group = 1:10,
#' dims = 10,
#' top = 500,
#' score = TRUE,
#' show_cols = TRUE,
#' type = "ggplot")
setMethod(f = "runAPL",
signature=(obj="matrix"),
function(obj,
group,
caobj = NULL,
dims = NULL,
nrow = 10,
top = 5000,
score = TRUE,
mark_rows = NULL,
mark_cols = NULL,
reps = 3,
python = FALSE,
row_labs = TRUE,
col_labs = TRUE,
type = "plotly",
show_cols = FALSE,
show_rows = TRUE,
score_cutoff = 0,
score_color = "rainbow",
pd_method = "elbow_rule",
pd_reps = 1,
pd_use = TRUE,
...) {
if (!is(obj, "matrix") & !is(obj, "dgCMatrix")) {
obj <- as.matrix(obj)
}
stopifnot(is(obj, "matrix") | is(obj, "dgCMatrix"))
run_APL(obj = obj,
caobj = caobj,
dims = dims,
group = group,
mark_rows = mark_rows,
mark_cols = mark_cols,
nrow = nrow,
top = top,
score = score,
reps = reps,
python = python,
row_labs = row_labs,
col_labs = col_labs,
type = type,
show_cols = show_cols,
show_rows = show_rows,
score_cutoff = score_cutoff,
score_color = score_color,
pd_method = pd_method,
pd_reps = pd_reps,
pd_use = pd_use)
})
#' @description
#' runAPL.SingleCellExperiment: Computes singular value decomposition and
#' coordinates for the Association Plot from SingleCellExperiment objects with
#' reducedDim(obj, "CA") slot (optional).
#'
#' @param assay Character. The assay from which extract the count matrix for
#' SVD, e.g. "RNA" for Seurat objects or "counts"/"logcounts" for
#' SingleCellExperiments.
#'
#' @rdname runAPL
#' @export
#' @examples
#'
#' ########################
#' # SingleCellExperiment #
#' ########################
#' library(SingleCellExperiment)
#' set.seed(1234)
#'
#' # Simulate counts
#' cnts <- mapply(function(x){rpois(n = 500, lambda = x)},
#' x = sample(1:100, 50, replace = TRUE))
#' rownames(cnts) <- paste0("gene_", 1:nrow(cnts))
#' colnames(cnts) <- paste0("cell_", 1:ncol(cnts))
#'
#' sce <- SingleCellExperiment(assays=list(counts=cnts))
#'
#' # (nonsensical) APL
#' runAPL(obj = sce,
#' group = 1:10,
#' dims = 10,
#' top = 500,
#' score = TRUE,
#' show_cols = TRUE,
#' type = "ggplot",
#' assay = "counts")
setMethod(f = "runAPL",
signature=(obj="SingleCellExperiment"),
function(obj,
group,
caobj = NULL,
dims = NULL,
nrow = 10,
top = 5000,
score = TRUE,
mark_rows = NULL,
mark_cols = NULL,
reps = 3,
python = FALSE,
row_labs = TRUE,
col_labs = TRUE,
type = "plotly",
show_cols = FALSE,
show_rows = TRUE,
score_cutoff = 0,
score_color = "rainbow",
pd_method = "elbow_rule",
pd_reps = 1,
pd_use = TRUE,
...,
assay = "counts"){
stopifnot("obj doesn't belong to class 'SingleCellExperiment'" =
is(obj, "SingleCellExperiment"))
mat <- SummarizedExperiment::assay(obj, assay)
if ("CA" %in% SingleCellExperiment::reducedDimNames(obj)){
caobj <- as.cacomp(obj, assay = assay)
}
if (!is(mat, "matrix") & !is(mat, "dgCMatrix")) {
mat <- as.matrix(mat)
}
run_APL(obj = mat,
caobj = caobj,
dims = dims,
group = group,
mark_rows = mark_rows,
mark_cols = mark_cols,
nrow = nrow,
top = top,
score = score,
reps = reps,
python = python,
row_labs = row_labs,
col_labs = col_labs,
type = type,
show_cols = show_cols,
show_rows = show_rows,
score_cutoff = score_cutoff,
score_color = score_color,
pd_method = pd_method,
pd_reps = pd_reps,
pd_use = pd_use)
})
#' @description
#' runAPL.Seurat: Computes singular value decomposition and coordinates for
#' the Association Plot from Seurat objects, optionally with a DimReduc Object
#' in the "CA" slot.
#'
#' @param assay Character. The assay from which extract the count matrix for
#' SVD, e.g. "RNA" for Seurat objects or "counts"/"logcounts" for
#' SingleCellExperiments.
#' @param slot character. The Seurat assay slot from which to extract the
#' count matrix.
#' @rdname runAPL
#' @export
#' @examples
#'
#' ###########
#' # Seurat #
#' ###########
#' library(Seurat)
#' set.seed(1234)
#'
#' # Simulate counts
#' cnts <- mapply(function(x){rpois(n = 500, lambda = x)},
#' x = sample(1:100, 50, replace = TRUE))
#' rownames(cnts) <- paste0("gene_", 1:nrow(cnts))
#' colnames(cnts) <- paste0("cell_", 1:ncol(cnts))
#'
#' seu <- CreateSeuratObject(counts = cnts)
#'
#' # (nonsensical) APL
#' runAPL(obj = seu,
#' group = 1:10,
#' dims = 10,
#' top = 500,
#' score = TRUE,
#' show_cols = TRUE,
#' type = "ggplot",
#' assay = "RNA",
#' slot = "counts")
setMethod(f = "runAPL",
signature=(obj="Seurat"),
function(obj,
group,
caobj = NULL,
dims = NULL,
nrow = 10,
top = 5000,
score = TRUE,
mark_rows = NULL,
mark_cols = NULL,
reps = 3,
python = FALSE,
row_labs = TRUE,
col_labs = TRUE,
type = "plotly",
show_cols = FALSE,
show_rows = TRUE,
score_cutoff = 0,
score_color = "rainbow",
pd_method = "elbow_rule",
pd_reps = 1,
pd_use = TRUE,
...,
assay = Seurat::DefaultAssay(obj),
slot = "counts"){
stopifnot("obj doesn't belong to class 'Seurat'" = is(obj, "Seurat"))
seu <- Seurat::GetAssayData(object = obj, assay = assay, slot = slot)
seu <- as.matrix(seu)
if ("CA" %in% Seurat::Reductions(obj)){
caobj <- as.cacomp(obj, assay = assay)
} else {
caobj <- NULL
}
run_APL(obj = seu,
caobj = caobj,
top = top,
dims= dims,
group = group,
score = score,
reps = reps,
python = python,
mark_rows = mark_rows,
mark_cols = mark_cols,
nrow = nrow,
row_labs = row_labs,
col_labs = col_labs,
type = type,
show_cols = show_cols,
show_rows = show_rows,
score_cutoff = score_cutoff,
score_color = score_color,
pd_method = pd_method,
pd_reps = pd_reps,
pd_use = pd_use)
})
#' @export
#' @rdname runAPL
#' @examples
#' set.seed(1234)
#'
#' # Simulate counts
#' cnts <- mapply(function(x){rpois(n = 500, lambda = x)},
#' x = sample(seq(0.01,0.1,by=0.01), 50, replace = TRUE))
#' rownames(cnts) <- paste0("gene_", 1:nrow(cnts))
#' colnames(cnts) <- paste0("cell_", 1:ncol(cnts))
#' cnts <- Matrix::Matrix(cnts)
#'
#' # (nonsensical) APL
#' runAPL(obj = cnts,
#' group = 1:10,
#' dims = 10,
#' top = 500,
#' score = TRUE,
#' show_cols = TRUE,
#' type = "ggplot")
setMethod(f = "runAPL",
signature=(obj="dgCMatrix"),
function(obj,
group,
caobj = NULL,
dims = NULL,
nrow = 10,
top = 5000,
score = TRUE,
mark_rows = NULL,
mark_cols = NULL,
reps = 3,
python = FALSE,
row_labs = TRUE,
col_labs = TRUE,
type = "plotly",
show_cols = FALSE,
show_rows = TRUE,
score_cutoff = 0,
score_color = "rainbow",
pd_method = "elbow_rule",
pd_reps = 1,
pd_use = TRUE,
...) {
stopifnot(is(obj, "dgCMatrix"))
run_APL(obj = obj,
caobj = caobj,
dims = dims,
group = group,
mark_rows = mark_rows,
mark_cols = mark_cols,
nrow = nrow,
top = top,
score = score,
reps = reps,
python = python,
row_labs = row_labs,
col_labs = col_labs,
type = type,
show_cols = show_cols,
show_rows = show_rows,
score_cutoff = score_cutoff,
score_color = score_color,
pd_method = pd_method,
pd_reps = pd_reps,
pd_use = pd_use)
})
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