Joint analysis of expression and variation at single cell resolution by scVar

Abstract

Tumor heterogeneity represents a critical determinant affecting both cancer diagnosis and therapeutic efficacy. Traditional bulk sequencing approaches are limited in their ability to resolve genomic alterations at the resolution of individual cells. In this study, we present scVar, an integrated analytical framework designed for mutation profiling using single-cell transcriptomic data. scVar enables sensitive and robust detection of single-nucleotide variants, particularly facilitating the identification of low-frequency variants. The framework incorporates customizable filtering parameters including variant allele frequency, and provides comprehensive functional and clinical annotations for identified mutations. Furthermore, scVar includes downstream analytical modules that facilitate the joint investigation of transcriptomic and mutational profiles within single cells. Benchmarking on simulated datasets and matched tumor samples demonstrates that scVar consistently outperforms existing methods, especially in detecting variants of low abundance, and shows strong concordance with whole-exome sequencing data. Application of scVar to non-small cell lung cancer samples effectively characterized ITH across spatially distinct tumor regions and diverse cellular populations. Collectively, scVar offers an integrated platform for concurrent analysis of somatic mutations and transcriptomic data derived from single-cell RNA sequencing, providing a powerful tool for elucidating tumor evolution and the complex interplay between genomic heterogeneity and the tumor microenvironment.