Cell type-specific weighting-factors to solve solid organs-specific limitations of single cell RNA-sequencing

PLoS Genet. 2024 Nov 18;20(11):e1011436. doi: 10.1371/journal.pgen.1011436. eCollection 2024 Nov.

Abstract

While single-cell RNA-sequencing (scRNA-seq) is a popular method to analyze gene expression and cellular composition at single-cell resolution, it harbors shortcomings: The failure to account for cell-to-cell variations of transcriptome-size (i.e., the total number of transcripts per cell) and also cell dissociation/processing-induced cryptic gene expression. This is particularly a problem when analyzing highly heterogeneous solid tissues/organs, which requires cell dissociation for the analysis. As a result, there exists a discrepancy between bulk RNA-seq result and virtually reconstituted bulk RNA-seq result using its composite scRNA-seq data. To fix this problem, we propose a computationally calculated coefficient, "cell type-specific weighting-factor (cWF)". Here, we introduce a concept and a method of its computation and report cWFs for 76 cell-types across 10 solid organs. Their fidelity is validated by more accurate reconstitution and deconvolution of bulk RNA-seq data of diverse solid organs using the scRNA-seq data and the cWFs of their composite cells. Furthermore, we also show that cWFs effectively predict aging-progression, implicating their diagnostic applications and also their association with aging mechanism. Our study provides an important method to solve critical limitations of scRNA-seq analysis of complex solid tissues/organs. Furthermore, our findings suggest a diagnostic utility and biological significance of cWFs.

MeSH terms

  • Aging / genetics
  • Animals
  • Gene Expression Profiling / methods
  • Humans
  • Mice
  • Organ Specificity / genetics
  • RNA-Seq / methods
  • Sequence Analysis, RNA* / methods
  • Single-Cell Analysis* / methods
  • Transcriptome* / genetics

Grants and funding

This work was funded by Innovative Science and Technology Initiative for Security Grant Number JPJ004596 ATLA Japan (T.N.S.), JST ERATO Grant Number JPMJER1303 Japan (T.N.S.), Nakatani Foundation (T.N.S.). K.T., S.K., and T.N.S. received salaries from Innovative Science and Technology Initiative for Security Grant Number JPJ004596 ATLA Japan, JST ERATO Grant Number JPMJER1303 Japan, and Nakatani Foundation. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.