CaSSiDI: Novel single-cell “Cluster Similarity Scoring and Distinction Index” reveals critical functions for PirB and context-dependent Cebpb repression

PirB is an inhibitory cell surface receptor particularly prominent on myeloid cells. PirB curtails the phenotypes of activated macrophages during inflammation or tumorigenesis, but its functions in macrophage homeostasis are obscure. To elucidate PirB-related functions in macrophages at steady-state, we generated and compared single-cell RNA-sequencing (scRNAseq) datasets obtained from myeloid cell subsets of wild type (WT) and PirB-deficient knockout (PirB KO) mice. To facilitate this analysis, we developed a novel approach to clustering parameter optimization called “Cluster Similarity Scoring and Distinction Index” (CaSSiDI). Our application of CaSSiDI in this study has revealed previously unknown roles for PirB in myeloid cell populations. In particular, we have discovered homeostatic functions for PirB that are related to Cebpb expression in distinct macrophage subsets. Our results establish the utility of CaSSiDI for single-cell assay analyses and the determination of optimal clustering parameters. Comparative analysis of scRNA-seq data obtained from tissue-derived macrophage subsets to investigate differential cluster formation in WT and PirB-KO samples. A new analysis pipeline "CaSSiDI" was established for the analysis.