estimateDispersions,DESeqDataSet-method {DESeq2} | R Documentation

## Estimate the dispersions for a DESeqDataSet

### Description

This function obtains dispersion estimates for Negative Binomial distributed data.

### Usage

    ## S4 method for signature 'DESeqDataSet'
    estimateDispersions(
      object,
      fitType = c("parametric", "local", "mean", "glmGamPoi"),
      maxit = 100,
      useCR = TRUE,
      weightThreshold = 0.01,
      quiet = FALSE,
      modelMatrix = NULL,
      minmu = if (fitType == "glmGamPoi") 1e-06 else 0.5
    )

### Arguments

`object` | a DESeqDataSet
`fitType` | either "parametric", "local", "mean", or "glmGamPoi" for the type of fitting of dispersions to the mean intensity. parametric - fit a dispersion-mean relation of the form: dispersion = asymptDisp + extraPois / mean via a robust gamma-family GLM. The coefficients `asymptDisp` and `extraPois` are given in the attribute `coefficients` of the `dispersionFunction` of the object. local - use the locfit package to fit a local regression of log dispersions over log base mean (normal scale means and dispersions are input and output for `dispersionFunction`). The points are weighted by normalized mean count in the local regression. mean - use the mean of gene-wise dispersion estimates. glmGamPoi - use the glmGamPoi package to fit the gene-wise dispersion, its trend and calculate the MAP based on the quasi-likelihood framework. The trend is calculated using a local median regression.
`maxit` | control parameter: maximum number of iterations to allow for convergence
`useCR` | whether to use Cox-Reid correction - see McCarthy et al (2012)
`weightThreshold` | threshold for subsetting the design matrix and GLM weights for calculating the Cox-Reid correction
`quiet` | whether to print messages at each step
`modelMatrix` | an optional matrix which will be used for fitting the expected counts. by default, the model matrix is constructed from `design(object)`
`minmu` | lower bound on the estimated count for fitting gene-wise dispersion

### Details

Typically the function is called with the idiom:

`dds <- estimateDispersions(dds)`

The fitting proceeds as follows: for each gene, an estimate of the dispersion is found which maximizes the Cox Reid-adjusted profile likelihood (the methods of Cox Reid-adjusted profile likelihood maximization for estimation of dispersion in RNA-Seq data were developed by McCarthy, et al. (2012), first implemented in the edgeR package in 2010); a trend line capturing the dispersion-mean relationship is fit to the maximum likelihood estimates; a normal prior is determined for the log dispersion estimates centered on the predicted value from the trended fit with variance equal to the difference between the observed variance of the log dispersion estimates and the expected sampling variance; finally maximum a posteriori dispersion estimates are returned. This final dispersion parameter is used in subsequent tests. The final dispersion estimates can be accessed from an object using `dispersions`. The fitted dispersion-mean relationship is also used in `varianceStabilizingTransformation`. All of the intermediate values (gene-wise dispersion estimates, fitted dispersion estimates from the trended fit, etc.) are stored in `mcols(dds)`, with information about these columns in `mcols(mcols(dds))`.

The log normal prior on the dispersion parameter has been proposed by Wu, et al. (2012) and is also implemented in the DSS package.

In DESeq2, the dispersion estimation procedure described above replaces the different methods of dispersion from the previous version of the DESeq package.

Since version 1.29, DESeq2 can call the glmGamPoi package, which can speed up the inference and is optimized for fitting many samles with very small counts (for example single cell RNA-seq data). To call functions from the glmGamPoi package, make sure that it is installed and set `fitType = "glmGamPoi"`. In addition, to the gene estimates, the trend and the MAP, the glmGamPoi package calculates the corresponding quasi-likelihood estimates. Those can be used with the `nbinomLRT()` test to get more precise p-value estimates.

The lower-level functions called by `estimateDispersions` are: `estimateDispersionsGeneEst`, `estimateDispersionsFit`, and `estimateDispersionsMAP`.

### Value

The DESeqDataSet passed as parameters, with the dispersion information filled in as metadata columns, accessible via `mcols`, or the final dispersions accessible via `dispersions`.

### References

- Simon Anders, Wolfgang Huber: Differential expression analysis for sequence count data. Genome Biology 11 (2010) R106, http://dx.doi.org/10.1186/gb-2010-11-10-r106

- McCarthy, DJ, Chen, Y, Smyth, GK: Differential expression analysis of multifactor RNA-Seq experiments with respect to biological variation. Nucleic Acids Research 40 (2012), 4288-4297, http://dx.doi.org/10.1093/nar/gks042

- Wu, H., Wang, C. & Wu, Z. A new shrinkage estimator for dispersion improves differential expression detection in RNA-seq data. Biostatistics (2012). http://dx.doi.org/10.1093/biostatistics/kxs033

- Ahlmann-Eltze, C., Huber, W. glmGamPoi: Fitting Gamma-Poisson Generalized Linear Models on Single Cell Count Data. Bioinformatics (2020). https://doi.org/10.1093/bioinformatics/btaa1009

### Examples

    dds <- makeExampleDESeqDataSet()
    dds <- estimateSizeFactors(dds)
    dds <- estimateDispersions(dds)
    head(dispersions(dds))

[Package DESeq2 version 1.32.0 Index]