--- title: "Word Cloud Annotation" author: "Zuguang Gu (z.gu@dkfz.de)" date: '`r Sys.Date()`' output: rmarkdown::html_vignette: fig_caption: true css: main.css vignette: > %\VignetteIndexEntry{Word Cloud Annotation} %\VignetteEngine{knitr::rmarkdown} %\VignetteEncoding{UTF-8} --- ```{r, echo = FALSE} library(knitr) knitr::opts_chunk$set( error = FALSE, tidy = FALSE, message = FALSE, warning = FALSE, fig.align = "center", dev = "jpeg" ) options(width = 100) ``` In the plot generated by `simplifyGO()`, there is a word cloud annotation attached to the heatmap which shows the general biological functions of the GO terms in each cluster. In this vignette, I will demonstrate a general function `anno_word_cloud()` that generates word cloud annotations to work with the **ComplexHeatmap** package. The `anno_word_cloud()` function basically a wrapper of two components: 1. constructing the word cloud (`word_cloud_grob()`) and 2. constructing an annotation (with `ComplexHeatmap::anno_link()`) that can be used in `rowAnnotation()` function. `anno_word_cloud()` has two main arguments `align_to` and `term`. `align_to` defines how to align the annotation to the heatmap. Similar as in `ComplexHeatmap::anno_link()`, the value of `align_to` can be a list of row indices where each index vector in the list corresponds to a word cloud. The value of `align_to` can also be a categorical vector where rows with the same level correspond to a same word cloud. If `align_to` is a categorical vector and `term` is a list, names of `term` should overlap to the levels in `align_to`. When `align_to` is set as a categorical vector, normally the same value is set to `row_split` in the main heatmap so that each row slice can correspond to a word cloud. `term` defines the description texts used for constructing the word clouds. The value should have the same format as `align_to`. If `align_to` is a list, `term` should also be a list. In this case, the length of vectors in `term` is not necessarily the same as in `align_to`. E.g. `length(term[[1]])` is not necessarily equal to `length(align_to[[1]]`. In other words, `term[[i]]` can contain arbitrary text as long as `length(term) == length(align_to)`. If `align_to` is a categorical vector, `term` should only be a character vector with the same length as `align_to`. Other arguments in `anno_word_cloud()` are straightforward to understand: - `exclude_words`: The words excluded from word cloud. - `max_words`: Maximal number of words in each word cloud. - `word_cloud_grob_param`: Graphic parameters send to `word_cloud_grob()`. The value should be a named list. - `fontsize_range`: Range of the font size. The value is a vector of length two. - `bg_gp`: Graphic parameters for controlling the background. - `side`: Side of the annotation relative to the heatmap. The value should be either "right" or "left". In the first example, I generate 10 word clouds and attach to the heatmap which is split into 10 groups by rows. ```{r, fig.width = 7, fig.height = 4} library(tm) data(crude) term = lapply(crude$content, function(x) x$content)[1:10] mat = matrix(rnorm(100*10), nrow = 100) split = sample(letters[1:10], 100, replace = TRUE) names(term) = letters[1:10] ## names of `term` must be the same as `unique(split)` str(term, nchar.max = 80) library(ComplexHeatmap) library(simplifyEnrichment) Heatmap(mat, row_split = split, right_annotation = rowAnnotation(wc = anno_word_cloud(split, term)) ) ``` The value for the first argument of `anno_word_cloud()` can also be explicitly converted into a list: ```{r, eval = FALSE} align_to = split(seq_along(split), split) Heatmap(mat, row_split = split, right_annotation = rowAnnotation(wc = anno_word_cloud(align_to, term)) ) ``` Argument `side` can be set to `"left"` to put the annotation on the left of the heatmap: ```{r, fig.width = 7, fig.height = 4} Heatmap(mat, row_split = split, row_dend_side = "right", row_title_side = "right", left_annotation = rowAnnotation(wc = anno_word_cloud(split, term, side = "left")) ) ``` In previous examples, font sizes are mapped to word frequencies. To make it more general, `term` can be set to a list of "keywords data frames" where each data frame has two columns: a column for keywords and a column for numeric values that will be mapped to font sizes of keywords. ```{r, fig.width = 7, fig.height = 4} term = lapply(letters[1:10], function(x) { data.frame(LETTERS[1:10], 1:10) }) names(term) = letters[1:10] Heatmap(mat, row_split = split, right_annotation = rowAnnotation(wc = anno_word_cloud(split, term)) ) ``` Specifically for GO terms, users do not need to provide the full GO descriptions, instead, they can only provide the GO IDs and the descriptions will be automatically extracted internally. In this case, users can use the helper function `anno_word_cloud_from_GO()` and set the GO ID list via the `go_id` argument. The format of `go_id` is similar as `term` in `anno_word_cloud()`, either a list of GO IDs or as a vector. Again note, if `go_id` is a list, e.g. `length(go_id[[1]])` is not necessarily equal to `length(align_to[[1]]`. To make the word clouds more meaningful, additionally enrichment via Fisher's Exact test is applied on keywords compared to background GO vocabulary. The significance of enrichment corresponds to the keyword font size. The next example visualizes an gene expression matrix where rows are split into three groups by k-means clustering. GO enrichment analysis was applied to the genes in the three groups separately. Variable `km` contains the k-means classification. `go_list` contains list of IDs of significant GO terms. ```{r} load(system.file("extdata", "golub_sig_go.RData", package = "simplifyEnrichment")) head(km) str(go_list) ``` Just make sure names of `go_list` should correspond to the levels in `km`. Adding word cloud annotations for the enriched GO terms is very straightforward: ```{r, echo = FALSE} ht_opt$message = FALSE ``` ```{r, fig.width = 7, fig.height = 4} library(circlize) Heatmap(t(scale(t(sig_mat))), name = "z-score", col = colorRamp2(c(-2, 0, 2), c("green", "white", "red")), show_row_names = FALSE, show_column_names = FALSE, row_title = NULL, column_title = NULL, show_row_dend = FALSE, show_column_dend = FALSE, row_split = km) + rowAnnotation(go = anno_word_cloud_from_GO(km, go_list, max_words = 30)) ``` Note you can still set `stat = "count"` to map word frequency to the font size of keywords. ```{r, fig.width = 7, fig.height = 4} Heatmap(t(scale(t(sig_mat))), name = "z-score", col = colorRamp2(c(-2, 0, 2), c("green", "white", "red")), show_row_names = FALSE, show_column_names = FALSE, row_title = NULL, column_title = NULL, show_row_dend = FALSE, show_column_dend = FALSE, row_split = km) + rowAnnotation(go = anno_word_cloud_from_GO(km, go_list, max_words = 30, stat = "count")) ``` Now there might be users who want to say: I don't actually need the word clouds, while I simply want to list key pathways in each cluster. This can be treated as a special case of word cloud where each word (or in general "a phrase as a word") has value of 1 associated. Users need to specify several more arguments in `anno_word_cloud()`, as shown in the following example. Here `add_new_line` controls whether to list each sentence in separated lines. `value_range` and `fontsize_range` controls the size of the text. Note the defaults of these two arguments work well when font sizes are mapped to word frequencies or significance from keywords enrichment, but if the values are of other types, you might need to manually reset them. ```{r, fig.width = 7, fig.height = 4} mat = matrix(rnorm(100*10), nrow = 100) split = sample(letters[1:5], 100, replace = TRUE) term = lapply(letters[1:5], function(x) { data.frame(c("This is the first sentence.", "This is the second sentence."), c(1, 1)) }) names(term) = letters[1:5] Heatmap(mat, row_split = split, right_annotation = rowAnnotation(wc = anno_word_cloud(split, term, add_new_line = TRUE, value_range = c(1, 1), fontsize_range = c(10, 10))) ) ``` The sentences can also have different font size, just set values in the second column of the data frames in `term`: ```{r, fig.width = 7, fig.height = 4} term = lapply(letters[1:5], function(x) { data.frame(c("This is the first sentence.", "This is the second sentence."), c(1, 2)) }) names(term) = letters[1:5] Heatmap(mat, row_split = split, right_annotation = rowAnnotation(wc = anno_word_cloud(split, term, add_new_line = TRUE, value_range = c(1, 2), fontsize_range = c(8, 12))) ) ```