Title: | Interactive Differential Expression AnaLysis |
---|---|
Description: | This package provides functions for an Interactive Differential Expression AnaLysis of RNA-sequencing datasets, to extract quickly and effectively information downstream the step of differential expression. A Shiny application encapsulates the whole package. Support for reproducibility of the whole analysis is provided by means of a template report which gets automatically compiled and can be stored/shared. |
Authors: | Federico Marini [aut, cre] |
Maintainer: | Federico Marini <[email protected]> |
License: | MIT + file LICENSE |
Version: | 2.1.1 |
Built: | 2024-12-20 03:38:19 UTC |
Source: | https://github.com/bioc/ideal |
Functions that are on their way to the function afterlife. Their successors are also listed.
... |
Ignored arguments. |
The successors of these functions are likely coming after the rework that
led to the creation of the mosdef
package. See more into its
documentation for more details.
All functions throw a warning, with a deprecation message pointing towards its descendent (if available).
goseqTable()
is now being replaced by the more flexible
mosdef::run_goseq()
function (which is even faster)
ggplotCounts()
is now being replaced by the more flexible, better
designed, and actually even more good looking mosdef::gene_plot()
function, with better default behavior and all.
deseqresult2tbl()
and deseqresult2DEgenes()
are now replaced by the
more flexible mosdef::deresult_to_df()
The internally defined functions createLinkENS()
, createLinkGeneSymbol()
,
and createLinkGO()
are now replaced by the equivalent functions in mosdef
:
mosdef::create_link_ENSEMBL()
, mosdef::create_link_NCBI()
and
mosdef::create_link_GO()
. Notably, the mosdef
package expanded on the
concept of automatically generated buttons, taking this to the extreme of
efficiency with the mosdef::buttonifier()
function
Federico Marini
# try(goseqtable())
# try(goseqtable())
Generate a tidy table with the DE genes from the results of DESeq
deseqresult2DEgenes(deseqresult, FDR = 0.05)
deseqresult2DEgenes(deseqresult, FDR = 0.05)
deseqresult |
A |
FDR |
Numeric value, the significance level for thresholding adjusted p-values |
A "tidy" data.frame with only genes marked as differentially expressed
# with simulated data... library(DESeq2) dds <- DESeq2::makeExampleDESeqDataSet(n = 100, m = 8, betaSD = 2) dds <- DESeq(dds) res <- results(dds) deseqresult2DEgenes(res)
# with simulated data... library(DESeq2) dds <- DESeq2::makeExampleDESeqDataSet(n = 100, m = 8, betaSD = 2) dds <- DESeq(dds) res <- results(dds) deseqresult2DEgenes(res)
Generate a tidy table with the results of DESeq
deseqresult2tbl(deseqresult)
deseqresult2tbl(deseqresult)
deseqresult |
A |
A "tidy" data.frame with all genes
# with simulated data... library(DESeq2) dds <- DESeq2::makeExampleDESeqDataSet(n = 100, m = 8, betaSD = 1) dds <- DESeq2::DESeq(dds) res <- DESeq2::results(dds) deseqresult2tbl(res)
# with simulated data... library(DESeq2) dds <- DESeq2::makeExampleDESeqDataSet(n = 100, m = 8, betaSD = 1) dds <- DESeq2::DESeq(dds) res <- DESeq2::results(dds) deseqresult2tbl(res)
Plot for normalized counts of a single gene, with jittered points superimposed on the boxplot
ggplotCounts( dds, gene, intgroup = "condition", annotation_obj = NULL, transform = TRUE, labels_repel = TRUE )
ggplotCounts( dds, gene, intgroup = "condition", annotation_obj = NULL, transform = TRUE, labels_repel = TRUE )
dds |
A |
gene |
A character, specifying the name of the gene to plot |
intgroup |
Interesting groups: a character vector of
names in |
annotation_obj |
A |
transform |
Logical value, corresponding whether to have log scale y-axis or not. Defaults to TRUE. |
labels_repel |
Logical value. Whether to use |
Note: this function relies on the DESeq2::plotCounts()
function of DESeq2,
therefore pseudocounts of 0.5 are added to each point
An object created by ggplot
library("airway") data("airway", package = "airway") airway dds_airway <- DESeq2::DESeqDataSetFromMatrix(assay(airway), colData = colData(airway), design = ~ cell + dex ) ggplotCounts(dds_airway, gene = "ENSG00000103196", # CRISPLD2 in the original publication intgroup = "dex" )
library("airway") data("airway", package = "airway") airway dds_airway <- DESeq2::DESeqDataSetFromMatrix(assay(airway), colData = colData(airway), design = ~ cell + dex ) ggplotCounts(dds_airway, gene = "ENSG00000103196", # CRISPLD2 in the original publication intgroup = "dex" )
A wrapper for extracting functional GO terms enriched in a list of (DE) genes, based on the algorithm and the implementation in the goseq package
goseqTable( de.genes, assayed.genes, genome = "hg38", id = "ensGene", testCats = c("GO:BP", "GO:MF", "GO:CC"), FDR_GO_cutoff = 1, nTop = 200, orgDbPkg = "org.Hs.eg.db", addGeneToTerms = TRUE )
goseqTable( de.genes, assayed.genes, genome = "hg38", id = "ensGene", testCats = c("GO:BP", "GO:MF", "GO:CC"), FDR_GO_cutoff = 1, nTop = 200, orgDbPkg = "org.Hs.eg.db", addGeneToTerms = TRUE )
de.genes |
A vector of (differentially expressed) genes |
assayed.genes |
A vector of background genes, e.g. all (expressed) genes in the assays |
genome |
A string identifying the genome that genes refer to, as in the
|
id |
A string identifying the gene identifier used by genes, as in the
|
testCats |
A vector specifying which categories to test for over representation amongst DE genes - can be any combination of "GO:CC", "GO:BP", "GO:MF" & "KEGG" |
FDR_GO_cutoff |
Numeric value for subsetting the results |
nTop |
Number of categories to extract, and optionally process for adding genes to the respective terms |
orgDbPkg |
Character string, named as the |
addGeneToTerms |
Logical, whether to add a column with all genes annotated to each GO term |
Note: the feature length retrieval is based on the goseq::goseq()
function,
and requires that the corresponding TxDb packages are installed and available
A table containing the computed GO Terms and related enrichment scores
library("airway") data("airway", package = "airway") airway dds_airway <- DESeq2::DESeqDataSetFromMatrix(assay(airway), colData = colData(airway), design = ~ cell + dex ) dds_airway <- DESeq2::DESeq(dds_airway) res_airway <- DESeq2::results(dds_airway) res_subset <- mosdef::deresult_to_df(res_airway)[1:100, ] myde <- res_subset$id myassayed <- rownames(res_airway) ## Not run: mygo <- goseqTable(myde, myassayed, testCats = "GO:BP", addGeneToTerms = FALSE ) head(mygo) ## End(Not run)
library("airway") data("airway", package = "airway") airway dds_airway <- DESeq2::DESeqDataSetFromMatrix(assay(airway), colData = colData(airway), design = ~ cell + dex ) dds_airway <- DESeq2::DESeq(dds_airway) res_airway <- DESeq2::results(dds_airway) res_subset <- mosdef::deresult_to_df(res_airway)[1:100, ] myde <- res_subset$id myassayed <- rownames(res_airway) ## Not run: mygo <- goseqTable(myde, myassayed, testCats = "GO:BP", addGeneToTerms = FALSE ) head(mygo) ## End(Not run)
ideal makes differential expression analysis interactive, easy and reproducible. This function launches the main application included in the package.
ideal( dds_obj = NULL, res_obj = NULL, annotation_obj = NULL, countmatrix = NULL, expdesign = NULL, gene_signatures = NULL )
ideal( dds_obj = NULL, res_obj = NULL, annotation_obj = NULL, countmatrix = NULL, expdesign = NULL, gene_signatures = NULL )
dds_obj |
A |
res_obj |
A |
annotation_obj |
A |
countmatrix |
A count matrix, with genes as rows and samples as columns. If not provided, it is possible to upload the data during the execution of the Shiny App |
expdesign |
A |
gene_signatures |
A list of vectors, one for each pathway/signature. This
is for example the output of the |
A Shiny App is launched for interactive data exploration and differential expression analysis
# with simulated data... library("DESeq2") dds <- DESeq2::makeExampleDESeqDataSet(n = 100, m = 8) cm <- counts(dds) cd <- colData(dds) # with the well known airway package... library("airway") data("airway", package = "airway") airway dds_airway <- DESeq2::DESeqDataSetFromMatrix(assay(airway), colData = colData(airway), design = ~ cell + dex ) ## Not run: ideal() ideal(dds) ideal(dds_airway) dds_airway <- DESeq2::DESeq(dds_airway) res_airway <- DESeq2::results(dds_airway) ideal(dds_airway, res_airway) ## End(Not run)
# with simulated data... library("DESeq2") dds <- DESeq2::makeExampleDESeqDataSet(n = 100, m = 8) cm <- counts(dds) cd <- colData(dds) # with the well known airway package... library("airway") data("airway", package = "airway") airway dds_airway <- DESeq2::DESeqDataSetFromMatrix(assay(airway), colData = colData(airway), design = ~ cell + dex ) ## Not run: ideal() ideal(dds) ideal(dds_airway) dds_airway <- DESeq2::DESeq(dds_airway) res_airway <- DESeq2::results(dds_airway) ideal(dds_airway, res_airway) ## End(Not run)
ideal makes differential expression analysis interactive, easy and reproducible. The analysis of RNA-seq datasets is guided by the Shiny app as main component of the package, which also provides a wide set of functions to efficiently extract information from the existing data. The app can be also deployed on a Shiny server, to allow its usage without any installation on the user's side.
ideal makes differential expression analysis interactive, easy and reproducible. The analysis of RNA-seq datasets is guided by the Shiny app as main component of the package, which also provides a wide set of functions to efficiently extract information from the existing data. The app can be also deployed on a Shiny server, to allow its usage without any installation on the user's side.
Federico Marini [email protected], 2016-2017
Maintainer: Federico Marini [email protected]
Useful links:
Report bugs at https://github.com/federicomarini/ideal/issues
MA-plot from base means and log fold changes, in the ggplot2 framework, with additional support to annotate genes if provided.
plot_ma( res_obj, FDR = 0.05, point_alpha = 0.2, sig_color = "red", annotation_obj = NULL, draw_y0 = TRUE, hlines = NULL, title = NULL, xlab = "mean of normalized counts - log10 scale", ylim = NULL, add_rug = TRUE, intgenes = NULL, intgenes_color = "steelblue", labels_intgenes = TRUE, labels_repel = TRUE )
plot_ma( res_obj, FDR = 0.05, point_alpha = 0.2, sig_color = "red", annotation_obj = NULL, draw_y0 = TRUE, hlines = NULL, title = NULL, xlab = "mean of normalized counts - log10 scale", ylim = NULL, add_rug = TRUE, intgenes = NULL, intgenes_color = "steelblue", labels_intgenes = TRUE, labels_repel = TRUE )
res_obj |
A |
FDR |
Numeric value, the significance level for thresholding adjusted p-values |
point_alpha |
Alpha transparency value for the points (0 = transparent, 1 = opaque) |
sig_color |
Color to use to mark differentially expressed genes. Defaults to red |
annotation_obj |
A |
draw_y0 |
Logical, whether to draw the horizontal line at y=0. Defaults to TRUE. |
hlines |
The y coordinate (in absolute value) where to draw horizontal lines, optional |
title |
A title for the plot, optional |
xlab |
X axis label, defaults to "mean of normalized counts - log10 scale" |
ylim |
Vector of two numeric values, Y axis limits to restrict the view |
add_rug |
Logical, whether to add rug plots in the margins |
intgenes |
Vector of genes of interest. Gene symbols if a |
intgenes_color |
The color to use to mark the genes on the main plot. |
labels_intgenes |
Logical, whether to add the gene identifiers/names close to the marked plots |
labels_repel |
Logical, whether to use |
The genes of interest are to be provided as gene symbols if a symbol
column is provided in res_obj
, or else by using the identifiers specified
in the row names
An object created by ggplot
library("airway") data("airway", package = "airway") airway dds_airway <- DESeq2::DESeqDataSetFromMatrix(assay(airway), colData = colData(airway), design = ~ cell + dex ) # subsetting for quicker run, ignore the next two commands if regularly using the function gene_subset <- c( "ENSG00000103196", # CRISPLD2 "ENSG00000120129", # DUSP1 "ENSG00000163884", # KLF15 "ENSG00000179094", # PER1 rownames(dds_airway)[rep(c(rep(FALSE, 99), TRUE), length.out = nrow(dds_airway))] ) # 1% of ids dds_airway <- dds_airway[gene_subset, ] dds_airway <- DESeq2::DESeq(dds_airway) res_airway <- DESeq2::results(dds_airway) plot_ma(res_airway, FDR = 0.05, hlines = 1) plot_ma(res_airway, FDR = 0.1, intgenes = c( "ENSG00000103196", # CRISPLD2 "ENSG00000120129", # DUSP1 "ENSG00000163884", # KLF15 "ENSG00000179094" # PER1 ) )
library("airway") data("airway", package = "airway") airway dds_airway <- DESeq2::DESeqDataSetFromMatrix(assay(airway), colData = colData(airway), design = ~ cell + dex ) # subsetting for quicker run, ignore the next two commands if regularly using the function gene_subset <- c( "ENSG00000103196", # CRISPLD2 "ENSG00000120129", # DUSP1 "ENSG00000163884", # KLF15 "ENSG00000179094", # PER1 rownames(dds_airway)[rep(c(rep(FALSE, 99), TRUE), length.out = nrow(dds_airway))] ) # 1% of ids dds_airway <- dds_airway[gene_subset, ] dds_airway <- DESeq2::DESeq(dds_airway) res_airway <- DESeq2::results(dds_airway) plot_ma(res_airway, FDR = 0.05, hlines = 1) plot_ma(res_airway, FDR = 0.1, intgenes = c( "ENSG00000103196", # CRISPLD2 "ENSG00000120129", # DUSP1 "ENSG00000163884", # KLF15 "ENSG00000179094" # PER1 ) )
Volcano plot for log fold changes and log p-values in the ggplot2 framework, with additional support to annotate genes if provided.
plot_volcano( res_obj, FDR = 0.05, ylim_up = NULL, vlines = NULL, title = NULL, intgenes = NULL, intgenes_color = "steelblue", labels_intgenes = TRUE, labels_repel = TRUE )
plot_volcano( res_obj, FDR = 0.05, ylim_up = NULL, vlines = NULL, title = NULL, intgenes = NULL, intgenes_color = "steelblue", labels_intgenes = TRUE, labels_repel = TRUE )
res_obj |
A |
FDR |
Numeric value, the significance level for thresholding adjusted p-values |
ylim_up |
Numeric value, Y axis upper limits to restrict the view |
vlines |
The x coordinate (in absolute value) where to draw vertical lines, optional |
title |
A title for the plot, optional |
intgenes |
Vector of genes of interest. Gene symbols if a |
intgenes_color |
The color to use to mark the genes on the main plot. |
labels_intgenes |
Logical, whether to add the gene identifiers/names close to the marked plots |
labels_repel |
Logical, whether to use |
The genes of interest are to be provided as gene symbols if a symbol
column is provided in res_obj
, or else b< using the identifiers specified
in the row names
An object created by ggplot
library("airway") data("airway", package = "airway") airway dds_airway <- DESeq2::DESeqDataSetFromMatrix(assay(airway), colData = colData(airway), design = ~ cell + dex ) # subsetting for quicker run, ignore the next two commands if regularly using the function gene_subset <- c( "ENSG00000103196", # CRISPLD2 "ENSG00000120129", # DUSP1 "ENSG00000163884", # KLF15 "ENSG00000179094", # PER1 rownames(dds_airway)[rep(c(rep(FALSE, 99), TRUE), length.out = nrow(dds_airway))] ) # 1% of ids dds_airway <- dds_airway[gene_subset, ] dds_airway <- DESeq2::DESeq(dds_airway) res_airway <- DESeq2::results(dds_airway) plot_volcano(res_airway)
library("airway") data("airway", package = "airway") airway dds_airway <- DESeq2::DESeqDataSetFromMatrix(assay(airway), colData = colData(airway), design = ~ cell + dex ) # subsetting for quicker run, ignore the next two commands if regularly using the function gene_subset <- c( "ENSG00000103196", # CRISPLD2 "ENSG00000120129", # DUSP1 "ENSG00000163884", # KLF15 "ENSG00000179094", # PER1 rownames(dds_airway)[rep(c(rep(FALSE, 99), TRUE), length.out = nrow(dds_airway))] ) # 1% of ids dds_airway <- dds_airway[gene_subset, ] dds_airway <- DESeq2::DESeq(dds_airway) res_airway <- DESeq2::results(dds_airway) plot_volcano(res_airway)
Returns a list of pathways from a GMT file.
read_gmt(gmtfile)
read_gmt(gmtfile)
gmtfile |
A character value, containing the location of the GMT formatted file. It can also be a file found online |
A list of vectors, one for each pathway in the GMT file.
# this example reads in the freely available pathways from wikipathways ## Not run: mysigs <- read_gmt( "http://data.wikipathways.org/20240910/gmt/wikipathways-20240910-gmt-Homo_sapiens.gmt" ) head(mysigs) # see how the gene identifiers are encoded as ENTREZ id ## End(Not run)
# this example reads in the freely available pathways from wikipathways ## Not run: mysigs <- read_gmt( "http://data.wikipathways.org/20240910/gmt/wikipathways-20240910-gmt-Homo_sapiens.gmt" ) head(mysigs) # see how the gene identifiers are encoded as ENTREZ id ## End(Not run)
This function tries to guess which separator was used in a text delimited file
sepguesser(file, sep_list = c(",", "\t", ";", " "))
sepguesser(file, sep_list = c(",", "\t", ";", " "))
file |
The name of the file which the data are to be read from |
sep_list |
A vector containing the candidates for being identified as
separators. Defaults to |
A character value, corresponding to the guessed separator. One of "," (comma), "\t" (tab), ";" (semicolon)," " (whitespace)
sepguesser(system.file("extdata/design_commas.txt", package = "ideal")) sepguesser(system.file("extdata/design_semicolons.txt", package = "ideal")) sepguesser(system.file("extdata/design_spaces.txt", package = "ideal")) mysep <- sepguesser(system.file("extdata/design_tabs.txt", package = "ideal")) # to be used for reading in the same file, without having to specify the sep
sepguesser(system.file("extdata/design_commas.txt", package = "ideal")) sepguesser(system.file("extdata/design_semicolons.txt", package = "ideal")) sepguesser(system.file("extdata/design_spaces.txt", package = "ideal")) mysep <- sepguesser(system.file("extdata/design_tabs.txt", package = "ideal")) # to be used for reading in the same file, without having to specify the sep
Plot a heatmap for the selected gene signature on the provided data, with the possibility to compactly display also DE only genes
sig_heatmap( vst_data, my_signature, res_data = NULL, FDR = 0.05, de_only = FALSE, annovec, title = "", cluster_rows = TRUE, cluster_cols = FALSE, anno_colData = NULL, center_mean = TRUE, scale_row = FALSE )
sig_heatmap( vst_data, my_signature, res_data = NULL, FDR = 0.05, de_only = FALSE, annovec, title = "", cluster_rows = TRUE, cluster_cols = FALSE, anno_colData = NULL, center_mean = TRUE, scale_row = FALSE )
vst_data |
A |
my_signature |
A character vector, usually named, containing the genes which compose the gene signature |
res_data |
A |
FDR |
Numeric value between 0 and 1, the False Discovery Rate |
de_only |
Logical, whether to display only DE genes belonging to the pathway - defaults to FALSE |
annovec |
A named character vector, with the corresponding annotation across IDs |
title |
Character, title for the heatmap |
cluster_rows |
Logical, whether to cluster rows - defaults to TRUE |
cluster_cols |
Logical, whether to cluster column - defaults to FALSE. Recommended to be set to TRUE if de_only is also set to TRUE |
anno_colData |
Character vector, specifying the elements of the colData information to be displayed as a decoration of the heatmap. Can be a vector of any length, as long as these names are included as colData. Defaults to NULL, which would plot no annotation on the samples. |
center_mean |
Logical, whether to perform mean centering on the expression values. Defaults to TRUE, as it improves the general readability of the heatmap |
scale_row |
Logical, whether to perform row-based standardization of the expression values |
A plot based on the pheatmap
function
# with the well known airway package... library("airway") data("airway", package = "airway") airway dds_airway <- DESeq2::DESeqDataSetFromMatrix(assay(airway), colData = colData(airway), design = ~ cell + dex ) ## Not run: dds_airway <- DESeq2::DESeq(dds_airway) res_airway <- DESeq2::results(dds_airway) vst_airway <- DESeq2::vst(dds_airway) library(org.Hs.eg.db) annovec <- mapIds(org.Hs.eg.db, rownames(dds_airway), "ENTREZID", "ENSEMBL") mysignatures <- read_gmt( "http://data.wikipathways.org/20190210/gmt/wikipathways-20190210-gmt-Homo_sapiens.gmt" ) mysignature_name <- "Lung fibrosis%WikiPathways_20190210%WP3624%Homo sapiens" library(pheatmap) sig_heatmap(vst_airway, mysignatures[[mysignature_name]], res_data = res_airway, de_only = TRUE, annovec = annovec, title = mysignature_name, cluster_cols = TRUE ) ## End(Not run)
# with the well known airway package... library("airway") data("airway", package = "airway") airway dds_airway <- DESeq2::DESeqDataSetFromMatrix(assay(airway), colData = colData(airway), design = ~ cell + dex ) ## Not run: dds_airway <- DESeq2::DESeq(dds_airway) res_airway <- DESeq2::results(dds_airway) vst_airway <- DESeq2::vst(dds_airway) library(org.Hs.eg.db) annovec <- mapIds(org.Hs.eg.db, rownames(dds_airway), "ENTREZID", "ENSEMBL") mysignatures <- read_gmt( "http://data.wikipathways.org/20190210/gmt/wikipathways-20190210-gmt-Homo_sapiens.gmt" ) mysignature_name <- "Lung fibrosis%WikiPathways_20190210%WP3624%Homo sapiens" library(pheatmap) sig_heatmap(vst_airway, mysignatures[[mysignature_name]], res_data = res_airway, de_only = TRUE, annovec = annovec, title = mysignature_name, cluster_cols = TRUE ) ## End(Not run)
Combine data from a typical DESeq2 run
wrapup_for_iSEE(dds, res)
wrapup_for_iSEE(dds, res)
dds |
A |
res |
A |
Combines the DESeqDataSet input and DESeqResults into a SummarizedExperiment object, which can be readily explored with iSEE.
A typical usage would be after running the DESeq2 pipeline as specified in one of the workflows which include this package, e.g. in the context of the ideal package.
A SummarizedExperiment object, with raw counts, normalized counts, and variance-stabilizing transformed counts in the assay slots; and with colData and rowData extracted from the corresponding input parameters
# with simulated data... library(DESeq2) dds <- DESeq2::makeExampleDESeqDataSet(n = 10000, m = 8) dds <- DESeq(dds) res <- results(dds) se <- wrapup_for_iSEE(dds, res) # library(iSEE) # iSEE(se) ## Not run: # or with the well known airway package... library("airway") data("airway", package = "airway") airway dds_airway <- DESeq2::DESeqDataSetFromMatrix(assay(airway), colData = colData(airway), design = ~ cell + dex ) dds_airway <- DESeq2::DESeq(dds_airway) res_airway <- DESeq2::results(dds_airway) se_airway <- wrapup_for_iSEE(dds_airway, res_airway) # iSEE(se_airway) ## End(Not run)
# with simulated data... library(DESeq2) dds <- DESeq2::makeExampleDESeqDataSet(n = 10000, m = 8) dds <- DESeq(dds) res <- results(dds) se <- wrapup_for_iSEE(dds, res) # library(iSEE) # iSEE(se) ## Not run: # or with the well known airway package... library("airway") data("airway", package = "airway") airway dds_airway <- DESeq2::DESeqDataSetFromMatrix(assay(airway), colData = colData(airway), design = ~ cell + dex ) dds_airway <- DESeq2::DESeq(dds_airway) res_airway <- DESeq2::results(dds_airway) se_airway <- wrapup_for_iSEE(dds_airway, res_airway) # iSEE(se_airway) ## End(Not run)