Bnip3lb-driven mitophagy sustains expansion of the embryonic hematopoietic stem cell pool

Abstract

Embryonic hematopoietic stem and progenitor cells (HSPCs) have the unique ability to undergo rapid proliferation while maintaining multipotency, a clinically-valuable quality which currently cannot be replicated in vitro. Here, we show that embryonic HSPCs achieve this state by precise spatio-temporal regulation of reactive oxygen species (ROS) via Bnip3lb-associated developmentally-programmed mitophagy, a distinct autophagic regulatory mechanism from that of adult HSPCs. While ROS drives HSPC specification in the dorsal aorta, scRNAseq and live-imaging of Tg(ubi:mitoQC) zebrafish indicate that mitophagy initiates as HSPCs undergo endothelial-to-hematopoietic transition and colonize the caudal hematopoietic tissue (CHT). Knockdown of bnip3lb reduced mitophagy and HSPC numbers in the CHT by promoting myeloid-biased differentiation and apoptosis, which was rescued by anti-oxidant exposure. Conversely, induction of mitophagy enhanced both embryonic HSPC and lymphoid progenitor numbers. Significantly, mitophagy activation improved ex vivo functional capacity of hematopoietic progenitors derived from human-induced pluripotent stem cells (hiPSCs), enhancing serial-replating hematopoietic colony forming potential. HIGHLIGHTSO_LIROS promotes HSPC formation in the dorsal aorta but negatively affects maintenance thereafter. C_LIO_LIHSPCs colonizing secondary niches control ROS levels via Bnip3lb-directed mitophagy. C_LIO_LIMitophagy protects nascent HSPCs from ROS-associated apoptosis and maintains multipotency. C_LIO_LIInduction of mitophagy enhances long-term hematopoietic potential of iPSC-derived HSPCs. C_LI