Single-cell trajectory analysis reveals a melanoma-driven distinct hematopoietic response in murine spleen

Abstract

Abstract Hematopoietic stem and progenitor cells (HSPCs) are a rare population of precursor cells residing in bone marrow that replenish blood cells throughout adult life. Growing evidence suggests that tumor progression can interfere with normal hematopoiesis, skew the host system to undergo myeloid biased changes and cause extramedullary hematopoiesis (EMH) in organs such as the spleen. In this study, we investigated tumor-driven phenotypic and molecular alterations in HSPCs during EMH using single-cell multiomics. We used a B16-F10 melanoma mouse model which showed splenomegaly and alterations in the composition of the HSPCs in the spleen consistent with EMH. We then isolated four HSPC populations from the spleen and bone marrow of melanoma-burdened mice using the BD FACSMelody™ Cell Sorter and used the BD Rhapsody™ Single-Cell Analysis System for the downstream single-cell multiomics analysis. We utilized oligo-conjugated antibodies for NGS-based sample multiplexing (BD® Single-Cell Multiplexing Kit) and protein detection (BD® AbSeq Assay), along with whole transcriptome analysis for a comprehensive analysis of mRNA expression. We observed tumor-driven distinct hematopoietic response and alterations in developmental trajectories of HSPCs during EMH using the X-shift algorithm paired with single-cell force-directed layout visualization. Our findings highlight melanoma-driven perturbations in hematopoiesis by utilizing single-cell multiomics in a murine model. Class 1 Laser Product. For Research Use Only. Not for use in diagnostic or therapeutic procedures. BD-24477 (v1.0) 1220 BD, the BD Logo, FACSMelody, Rhapsody and AbSeq are trademarks of Becton, Dickinson and Company or its affiliates ©2020 BD. All rights reserved.