DRIMSeq: Differential transcript usage and tuQTL analyses with Dirichlet-multinomial model in RNA-seq

Description Usage Arguments Details Value Author(s) References See Also Examples

Obtain the maximum likelihood estimates of Dirichlet-multinomial (gene-level) and/or beta-binomial (feature-level) regression coefficients, feature proportions in each sample and corresponding likelihoods. In the differential exon/transcript usage analysis, the regression model is defined by a design matrix. In the exon/transcript usage QTL analysis, regression models are defined by genotypes. Currently, beta-binomial model is implemented only in the differential usage analysis.

dmFit(x, ...)

## S4 method for signature 'dmDSprecision'
dmFit(x, design, one_way = TRUE, bb_model = TRUE,
  prop_mode = "constrOptim", prop_tol = 1e-12, coef_mode = "optim",
  coef_tol = 1e-12, verbose = 0, BPPARAM = BiocParallel::SerialParam())

## S4 method for signature 'dmSQTLprecision'
dmFit(x, one_way = TRUE,
  prop_mode = "constrOptim", prop_tol = 1e-12, coef_mode = "optim",
  coef_tol = 1e-12, verbose = 0, BPPARAM = BiocParallel::SerialParam())

`x`	`dmDSprecision` or `dmSQTLprecision` object.
`...`	Other parameters that can be defined by methods using this generic.
`design`	Numeric matrix defining the full model.
`one_way`	Logical. Should the shortcut fitting be used when the design corresponds to multiple group comparison. This is a similar approach as in `edgeR`. If `TRUE` (the default), then proportions are fitted per group and regression coefficients are recalculated from those fits.
`bb_model`	Logical. Whether to perform the feature-level analysis using the beta-binomial model.
`prop_mode`	Optimization method used to estimate proportions. Possible value `"constrOptim"`.
`prop_tol`	The desired accuracy when estimating proportions.
`coef_mode`	Optimization method used to estimate regression coefficients. Possible value `"optim"`.
`coef_tol`	The desired accuracy when estimating regression coefficients.
`verbose`	Numeric. Definie the level of progress messages displayed. 0 - no messages, 1 - main messages, 2 - message for every gene fitting.
`BPPARAM`	Parallelization method used by `bplapply`.

In the regression framework here, we adapt the idea from glmFit in edgeR about using a shortcut algorithm when the design is equivalent to simple group fitting. In such a case, we estimate the DM proportions for each group of samples separately and then recalculate the DM (and/or the BB) regression coefficients corresponding to the design matrix. If the design matrix does not define a simple group fitting, for example, when it contains a column with continuous values, then the regression framework is used to directly estimate the regression coefficients.

Arguments that are used for the proportion estimation in each group when the shortcut fitting can be used start with prop_, and those that are used in the regression framework start with coef_.

In the differential transcript usage analysis, setting one_way = TRUE allows switching to the shortcut algorithm only if the design is equivalent to simple group fitting. one_way = FALSE forces usage of the regression framework.

In the QTL analysis, currently, genotypes are defined as numeric values 0, 1, and 2. When one_way = TRUE, simple multiple group fitting is performed. When one_way = FALSE, a regression framework is used with the design matrix defined by a formula ~ group where group is a continuous (not categorical) variable with values 0, 1, and 2.

Returns a dmDSfit or dmSQTLfit object.

Malgorzata Nowicka

McCarthy, DJ, Chen, Y, Smyth, GK (2012). Differential expression analysis of multifactor RNA-Seq experiments with respect to biological variation. Nucleic Acids Research 40, 4288-4297.

plotProportions glmFit

# --------------------------------------------------------------------------
# Create dmDSdata object 
# --------------------------------------------------------------------------
## Get kallisto transcript counts from the 'PasillaTranscriptExpr' package

library(PasillaTranscriptExpr)

data_dir  <- system.file("extdata", package = "PasillaTranscriptExpr")

## Load metadata
pasilla_metadata <- read.table(file.path(data_dir, "metadata.txt"), 
header = TRUE, as.is = TRUE)

## Load counts
pasilla_counts <- read.table(file.path(data_dir, "counts.txt"), 
header = TRUE, as.is = TRUE)

## Create a pasilla_samples data frame
pasilla_samples <- data.frame(sample_id = pasilla_metadata$SampleName, 
  group = pasilla_metadata$condition)
levels(pasilla_samples$group)

## Create a dmDSdata object
d <- dmDSdata(counts = pasilla_counts, samples = pasilla_samples)

## Use a subset of genes, which is defined in the following file
gene_id_subset <- readLines(file.path(data_dir, "gene_id_subset.txt"))

d <- d[names(d) %in% gene_id_subset, ]

# --------------------------------------------------------------------------
# Differential transcript usage analysis - simple two group comparison 
# --------------------------------------------------------------------------

## Filtering
## Check what is the minimal number of replicates per condition 
table(samples(d)$group)

d <- dmFilter(d, min_samps_gene_expr = 7, min_samps_feature_expr = 3,
  min_gene_expr = 10, min_feature_expr = 10)

plotData(d)

## Create the design matrix
design_full <- model.matrix(~ group, data = samples(d))

## To make the analysis reproducible
set.seed(123)
## Calculate precision
d <- dmPrecision(d, design = design_full)

plotPrecision(d)

head(mean_expression(d))
common_precision(d)
head(genewise_precision(d))

## Fit full model proportions
d <- dmFit(d, design = design_full)

## Get fitted proportions
head(proportions(d))
## Get the DM regression coefficients (gene-level) 
head(coefficients(d))
## Get the BB regression coefficients (feature-level) 
head(coefficients(d), level = "feature")