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

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 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-renwal potential of MPPs thus confer long-term multilineage repopulating capacity on MPPs, offering promising means to involve MPPs to augment cell source in clinical transplantation settings. For MP sequencing, 0.1 million myeloid progenitor cells (Lin-ckit+Sca1-) were sorted as one sample from bone marrow nucleated cells of respective recipient mice. Donor-derived NA MP cells (Tdtomato+) were sorted from NA MPP recipient mice 3 months post transplantation, while WT MP cells(CD45.2+) were obtained from control group received WT MPP transplantation at the same time point. Following RNA extraction with RNeasy micro kit (QIAGEN), total RNA of each sorted myeloid progenitor sample was used for sequencing library preparation with illumina Truseq RNA Sample Preparation Kit (RS-122-2001). All libraries were sequenced by illumina sequencer NextSeq 500 (illumina).