CDSeq: A novel complete deconvolution method for dissecting heterogeneous samples using gene expression data

Quantifying cell-type proportions and their corresponding gene expression profiles in tissue samples would enhance understanding of the contributions of individual cell types to the physiological states of the tissue. Current approaches that address tissue heterogeneity have drawbacks. Experimental techniques, such as fluorescence-activated cell sorting, and single cell RNA sequencing are expensive. Computational approaches that use expression data from heterogeneous samples are promising, but most of the current methods estimate either cell-type proportions or cell-type-specific expression profiles by requiring the other as input. Although such partial deconvolution methods have been successfully applied to tumor samples, the additional input required may be unavailable. We introduce a novel complete deconvolution method, CDSeq, that uses only RNA-Seq data from bulk tissue samples to simultaneously estimate both cell-type proportions and cell-type-specific expression profiles. Using several synthetic and real experimental datasets with known cell-type composition and cell-type-specific expression profiles, we compared CDSeq’s complete deconvolution performance with seven other established deconvolution methods. Complete deconvolution using CDSeq represents a substantial technical advance over partial deconvolution approaches and will be useful for studying cell mixtures in tissue samples. CDSeq is available at GitHub repository (MATLAB and Octave code): https://github.com/kkang7/CDSeq.Author summary: Understanding the cellular composition of bulk tissues is critical to investigate the underlying mechanisms of many biological processes. Single cell sequencing is a promising technique, however, it is expensive and the analysis of single cell data is non-trivial. Therefore, tissue samples are still routinely processed in bulk. To estimate cell-type composition using bulk gene expression data, computational deconvolution methods are needed. Many deconvolution methods have been proposed, however, they often estimate only cell type proportions using a reference cell type gene expression profile, which in many cases may not be available. We present a novel complete deconvolution method that uses only bulk gene expression data to simultaneously estimate cell-type-specific gene expression profiles and sample-specific cell-type proportions. We showed that, using multiple RNA-Seq and microarray datasets where the cell-type composition was previously known, our method could accurately determine the cell-type composition. By providing a method that requires a single input to determine both cell-type proportion and cell-type-specific expression profiles, we expect that our method will be beneficial to biologists and facilitate the research and identification of mechanisms underlying many biological processes.