Erythroid differentiation intensifies RNA mis-splicing inSF3B1-mutant myelodysplastic syndromes with ring sideroblasts

Abstract Myelodysplastic syndromes with ring sideroblasts (MDS-RS) commonly originate from mutations in the splicing factor SF3B1 ( SF3B1 mt ). SF3B1 mt cause RNA mis-splicing, mechanistically established as the major driver of RS development. However, little is known about RS fate and biology after their initial formation in the human bone marrow. We here achieve isolation of viable RS from patient samples, enabling the first complete investigation of SF3B1 mt development from stem cell to RS. We show that RS skew MACS-isolated CD34 + data towards erythroid features not recapitulated in single-cell RNAseq. We demonstrate that RS divide, differentiate, enucleate and actively respond to mis-splicing/oxidative stress, decreasing wildtype stem cell fitness via GDF15 overproduction. We identify circulating RS as a uniform clinical feature associated with disease burden. Finally, we establish that SF3B1 mt mis-splicing intensifies during erythroid differentiation and demonstrate through combined transcriptomics/proteomics an uncoupling of RNA/protein biology in RS encompassing severe and dysfunctional mis-splicing of proapoptotic genes. Statement of significance We here combine a novel method for RS isolation with state-of-the-art multiomics to perform the first complete investigation of SF3B1 mt MDS-RS hematopoiesis from stem cell to RS. We identify the survival mechanisms underlying SF3B1 mt erythropoiesis and establish an active role for erythroid differentiation and RS themselves in SF3B1 mt MDS-RS pathogenesis.