Hematopoietic stem cell division is governed by distinct RUNX1 binding partners.

Robertson, A. L.; Yue, L.; Choudhuri, A.; Kubaczka, C.; Wattrus, S. J.; Mandelbaum, J.; Avagyan, S.; Yang, S.; Freeman, R. J.; Chan, V.; Blair, M. C.; Daley, G. Q.; Zon, L. I.

doi:10.1101/2024.06.07.596542

bioRxiv · 10.1101/2024.06.07.596542

Hematopoietic stem cell division is governed by distinct RUNX1 binding partners.

Robertson, A. L.·Yue, L.·Choudhuri, A.·Kubaczka, C.·Wattrus, S. J.·Mandelbaum, J.·Avagyan, S.·Yang, S.·Freeman, R. J.·Chan, V.·Blair, M. C.·Daley, G. Q.·Zon, L. I.

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Abstract

A defined number of hematopoietic stem cell (HSC) clones are born during development and expand to form the pool of adult stem cells. An intricate balance between self-renewal and differentiation of these HSCs supports hematopoiesis for life. HSC fate is determined by complex transcription factor networks that drive cell-type specific gene programs. The transcription factor RUNX1 is required for definitive hematopoiesis, and mutations in Runx1 have been shown to reduce clonal diversity. The RUNX1 cofactor, CBFy, stabilizes RUNX1 binding to DNA, and disruption of their interaction alters downstream gene expression. Chemical screening for modulators of Runx1 and HSC expansion in zebrafish led us to identify a new mechanism for the RUNX1 inhibitor, Ro5-3335. We found that Ro5-3335 increased HSC divisions in zebrafish, and animals transplanted with Ro5-3335 treated cells had enhanced chimerism compared to untreated cells. Using human CD34+ cells, we show that Ro5-3335 remodels the RUNX1 transcription complex by binding to ELF1, independent of CBFy. This allows specific expression of cell cycle and hematopoietic genes that enhance HSC self-renewal and prevent differentiation. Furthermore, we provide the first evidence to show that it is possible to pharmacologically increase the number of stem cell clones in vivo, revealing a previously unknown mechanism for enhancing clonal diversity. Our studies have revealed a mechanism by which binding partners of RUNX1 determine cell fate, with ELF transcription factors guiding cell division. This information could lead to treatments that enhance clonal diversity for blood diseases.

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Robertson, A. L., Yue, L., Choudhuri, A., Kubaczka, C., Wattrus, S. J., Mandelbaum, J., Avagyan, S., Yang, S., Freeman, R. J., Chan, V., Blair, M. C., Daley, G. Q., Zon, L. I.. 2024-06-08. Hematopoietic stem cell division is governed by distinct RUNX1 binding partners.. https://doi.org/10.1101/2024.06.07.596542

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