Project description:mRNA profiling of CD34+ human cord blood-derived cell treated with UM171, SR1 or both mRNA profiles of CD34+ human cord blood-derived cell treated with DMSO (control), SR1 [500nM], UM171 [35nM] or combination SR1 [500nM]+ UM171 [35nM] for 30min, 3hr, 12hr, 24hr, 48hr, 72hr were generated by deep sequencing
Project description:Ex vivo expansion of hematopoietic stem cells (HSC) requires the maintenance of a stemness state while cells are proliferating. This can be achieved via exposure to UM171 which leads to the degradation of chromatin modifiers and prevents the loss of key epigenetic marks. However, the chromatin landscape varies across populations within the hematopoietic system and the effect of UM171 on self-renewal and differentiation potential of different hematopoietic progenitor cells is less characterized. To address this, we used the CellTag barcoding approach to track the fate of individual stem and progenitor cells during in vitro expansion. We showed that in addition to its HSC self-renewing property, UM171 specifically modulates cell fate of a precursor common to erythroid, megakaryocytic, and mast cells in favor of self-renewal and a mast-bias differentiation trajectory. This differentiation bias can be driven by pro-inflammatory signaling pathways that are activated downstream of UM171 and resulted in an abundant mast cell population that can be transplanted as part of the graft to populate mice tissues in xenotransplantation studies.
Project description:RNASeq data for mPB or CB-derived CD34+ exposed to UM171 human mobilized peripheral blood or cord blood-derived CD34(+) cells were cultured for 16 hours with vehicle (DMSO), dose response of UM171 [11.9nM, 19nM, 30.5nM, 48.8nM, 78.1nM and 125nM], SR1 [500nM] and combination of( UM171 [48.8nM]+SR1 [500nM])
Project description:Elucidation of the molecular cues required to balance adult stem cell self-renewal and differentiation is critical for advancing cellular therapies. Herein, we report that the hematopoietic stem cell (HSC) self-renewal agonist UM171 triggers a balanced pro- and anti-inflammatory/detoxification network that relies on NFKB activation and protein C receptor-dependent ROS detoxification, respectively. We demonstrate that within this network, EPCR serves as a critical protective component as its deletion hypersensitizes primitive hematopoietic cells to pro-inflammatory signals and ROS accumulation resulting in compromised stem cell function. Conversely, abrogation of the pro-inflammatory activity of UM171 through treatment with dexamethasone, cAMP elevating agents or NFKB inhibitors abolishes EPCR upregulation and HSC expansion. Together, these results show that UM171 stimulates ex vivo HSC expansion by establishing a critical balance between key pro- and anti-inflammatory mediators of self-renewal.
Project description:Earlier we identified the protein degradation properties of small molecule UM171 (Subramaniam et al., 2020, Blood). Here we employed quantitative proteomics approach to map the global targets of UM171. Our study revealed multiple targets including the members of CoREST complex such as RCOR1 and LSD1.
Project description:Earlier we identified the protein degradation properties of small molecule UM171 (Subramaniam et al., 2020, Blood). Here we employed quantitative proteomics approach to map the global targets of UM171. Our study revealed multiple targets including the members of CoREST complex such as RCOR1 and LSD1.
