Enforced Expression of NUP98-HOXA10hd Fusion Gene in Multipotent Progenitors Support Long-term Hematopoiesis in primary mice [MPP]

Allogenic hematopoietic stem cell (HSC) transplantation is widely used for treatment of blood disorders to re-establish long-term multi-lineage hematopoiesis. Enhance self-renewal potential of progenitor population to support hematopoiesis offer an alternative and complementary approach to achieve similar or enhance therapeutic effects. Nup98-Hoxa10hd fusion gene (NA) has been shown to confer expansion, anti-stress response and engraftment competitiveness on HSC. However, whether the ectopic expression of NA in multipotent progenitors (MPPs) could enhance their self-renewal potential and confer long-term multi-lineage hematopoiesis remains unknown. In this study, we showed that ectopic expression in MPPs confer long-term multi-lineage hematopoiesis in recipient mice. We further showed that NA upregulated pathways of cell cycle regulation, epigenetic regulation and response to stress in MPPs. These molecular traits increased the self-renewal potential of NA MPPs, which resulting in production of lineage-maintaining committed progenitor cells. Transcriptome analysis of NA myeloid progenitors identified genes regulating hematopoiesis, homeostasis, phosphorylation. In summary, we show that ectopic expression of Nup98-Hoxa10hd fusion gene enhance self-renewal potential of MPPs thus confer long-term multi-lineage repopulating capacity on MPPs, offering promising means to involve MPPs to augment cell source in clinical transplantation settings. We sorted 1000 MPP (Lin-CD48-ckit+Sca1+CD135+CD150-) cells for each NA MPP replicate from bone marrow of NA mice. Equal number of WT MPP cells were sorted to generated WT MPP replicate from bone marrow of age-matched CD45.2 mice. Sequencing libraries of MPP were generated as previously described (Dong et al., 2019; Tang et al., 2010). Briefly, cDNA of sorted 1000 MPP aliquots were generated, amplified and used for sequencing library preparation with illumina Nextera XT DNA Sample Preparation Kit (FC-131-1096). All libraries were sequenced by illumina sequencer NextSeq 500 (illumina).