AA2P-mediated DNA demethylation synergizes with stem cell agonists to promote expansion of hematopoietic stem cells

Javed K Manesia; Harinad B Maganti; Sakhar Almoflehi; Suria Jahan; Tanvir Hasan; Roya Pasha; Chelsea McGregor; Nellie Dumont; Josée Laganière; Julie Audet; Nicolas Pineault

doi:10.1016/j.crmeth.2023.100663

AA2P-mediated DNA demethylation synergizes with stem cell agonists to promote expansion of hematopoietic stem cells

Cell Rep Methods. 2023 Dec 18;3(12):100663. doi: 10.1016/j.crmeth.2023.100663. Epub 2023 Dec 8.

Authors

Affiliations

¹ Canadian Blood Services, Centre for Innovation, Ottawa, ON, Canada.
² Canadian Blood Services, Centre for Innovation, Ottawa, ON, Canada; Biochemistry, Microbiology and Immunology Department, University of Ottawa, Ottawa, ON, Canada.
³ Héma-Québec, Innovation, Québec, QC, Canada.
⁴ Institute of Biomedical Engineering, University of Toronto, Toronto, ON, Canada.
⁵ Canadian Blood Services, Centre for Innovation, Ottawa, ON, Canada; Biochemistry, Microbiology and Immunology Department, University of Ottawa, Ottawa, ON, Canada. Electronic address: nicolas.pineault@blood.ca.

Abstract

Small molecules have enabled expansion of hematopoietic stem and progenitor cells (HSPCs), but limited knowledge is available on whether these agonists can act synergistically. In this work, we identify a stem cell agonist in AA2P and optimize a series of stem cell agonist cocktails (SCACs) to help promote robust expansion of human HSPCs. We find that SCACs provide strong growth-promoting activities while promoting retention and function of immature HSPC. We show that AA2P-mediated HSPC expansion is driven through DNA demethylation leading to enhanced expression of AXL and GAS6. Further, we demonstrate that GAS6 enhances the serial engraftment activity of HSPCs and show that the GAS6/AXL pathway is critical for robust HSPC expansion.

Keywords: AXL; CP: Stem cell; DNA demethylation; GAS6; engraftment; epigenetic modulators; hematopoietic stem cells; stem cell agonists; stem cell expansion; umbilical cord blood.

MeSH terms

Cells, Cultured
DNA Demethylation*
Hematopoietic Stem Cell Transplantation*
Hematopoietic Stem Cells / metabolism
Humans