Novel Assignment of Gene Markers to Hematological and Immune Cells Based on Single-Cell Transcriptomics

There are many different cells that perform highly specialized functions in the human hematological and immune systems. Due to the relevance of their activity, in this work we investigated the cell types and subtypes that form this complex system, using single-cell RNA sequencing (scRNA-seq) to dissect and assess the markers that best define each cell population. We first developed an optimized computational workflow for analyzing large scRNA-seq datasets. We then used it to find gene markers of the different cell types present in bone marrow (BM) and peripheral blood (PB). We analyzed three different single-cell datasets to find specific cell markers using this strategy: first, we searched in the CD marker genes and then in the genes encoding membrane proteins and finally in all detected protein-coding genes. This allowed us not only to confirm known CDs that best mark some cell types (e.g., monocytes, B cells, NK cells, etc.) but also to test the ability of new genes to distinguish specific cell types. Finally, we applied a machine learning method (Random Forest) to test the accuracy of the different markers we found. As a result of all this work, we have found and propose specific and robust gene signatures to identify different types and subtypes of hematological and immune cells.