Identification of a retinoic acid-dependent haemogenic endothelial progenitor from human pluripotent stem cells

Creamer, J. Philip; Valsoni, Sara; Dege, Carissa; Scarfò, Rebecca; Dacunto, Analisa; Cascione, Sara; Randolph, Lauren N.; Cavalca, Eleonora; Merelli, Ivan; Morris, Samantha A.; Ditadi, Andrea; Sturgeon, Christopher M.

doi:10.1038/s41556-022-00898-9

Cited by 22 publications

(27 citation statements)

References 41 publications

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“…Here, it primes the small pool of LT-HSCs for self-renewal and proliferation via reducing the levels of RA signaling in these cells. Of note, recent studies have established that all HSC activity is contained within RA metabolizing cells (Chanda et al, 2013; Schonberger et al, 2022; Luff et al, 2022). Although the main function of IκBα is the inactivation of NFκB complex by cytoplasmic retention or by active removal from the nucleus (Beg and Baldwin Jr., 1993), there is an ancestral function of IκBα that is evolutionary conserved and is directly affecting PRC2 complex activity on stem cell specific genes (Brena et al, 2020; Mulero et al, 2013).…”

Section: Discussionmentioning

confidence: 99%

IκBα controls dormancy induction in Hematopoietic stem cell development via retinoic acid

Thambyrajah

Fadlullah

Proffitt

et al. 2022

Preprint

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Recent findings are challenging the classical hematopoietic model in which long-term hematopoietic stem cells (LT-HSC) are the base of the hematopoietic system. Clonal dynamics analysis of the hematopoietic system indicate that LT-HSC are not the main contributors of normal hemapoiesis in physiological conditions and the hematopoietic system is mainly maintained by multipotent progenitors (MPPs, hereafter HPC) and LT-HSCs are mostly in a non-active state. The first HSCs emerge from the aorta-gonad and mesonephros (AGM) region along with hematopoietic progenitors (HPC) within hematopoietic clusters. Molecular pathways that determine the HSC fate instead of HPC are still unknown, although inflammatory signaling, including NF-ΚB has been implicated in the development of HSCs. Here, we identify a chromatin binding function for IΚBα (also known as the inhibitor of NF-ΚB) that is Polycomb repression complex 2 (PRC2)- dependent and specifically determines dormant vs proliferating HSCs from the onset of their emergence in the AGM. We find a specific reduction of LT-HSCs in the IΚBα knockout new-born pups. This defect is manifested at the FL stage already, and traceable to the first emerging HSCs in the E11.5 AGM, without affecting the general HPC population. IΚBα-deficient LT-HSCs express dormancy signature genes, are less proliferative and can robustly respond to activation stimuli such asin vitroculture and serial transplantation. At the molecular level, we find decreased PRC2-dependent H3K27me3 at the promoters of several retinoic acid signaling elements in the IΚBα- deficient aortic endothelium and E14.5 FL LT-HSCs. Additionally, IΚBα binding itself is found in the promoters of retinoic acid receptors rarα in the AGM, and rarγ in the LT-HSC of FL. Overall, we demonstrate that the retinoic acid pathway is over-activated in the hematopoietic clusters of IΚBα-deficient AGMs leading to premature dormancy of LT- HSCs that persists in the FL LT-HSCs.

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Section: Discussionmentioning

confidence: 99%

IκBα controls dormancy induction in Hematopoietic stem cell development via retinoic acid

Thambyrajah

Fadlullah

Proffitt

et al. 2022

Preprint

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“…These iPSCs can give rise to blood progenitors that recapitulate key pathological features of JMML, including increased myelopoiesis and constitutive GM‐CSF pathway activation 119 . Sophisticated strategies now enable generation and isolation of engraftable hematopoietic progenitors from iPSC cultures 121,122 . These techniques, coupled with relevant mouse models and primary JMML cells, should afford granular insights into how JMML cells of origin emerge and transform.…”

Section: Age‐restricted Childhood Leukemiasmentioning

confidence: 99%

Layered immunity and layered leukemogenicity: Developmentally restricted mechanisms of pediatric leukemia initiation

Mendoza-Castrejon

Magee

2023

Immunological Reviews

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Leukemia is the most common childhood cancer, accounting for more deaths due to disease in developed nations than any other illness. 1 While leukemias can occur at any age, the mutations that cause childhood leukemias often differ from those that cause adult leukemias. 2 Some types of leukemia, such as juvenile myelomonocytic leukemia, present almost exclusively in early childhood. 3 These observations suggest that for several leukemogenic mutations, transformation efficiency changes with age. Alternatively, age-restricted leukemias might arise from cells of origin that emerge, and then disappear, through the course of normal ontogeny. In either case, transcriptional programs that direct normal ontogeny of the immune system can substantially influence leukemogenesis.

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“…Based on our results obtained with the human embryos, we next investigated whether CD32 can be a reliable marker for isolating HECs from hPSC differentiating cultures. We monitored its expression in WNTd intra-embryonic-like HOXA + HECs 27, 30, 32, 39 (Extended Data Fig. 3a).…”

Section: Mainmentioning

confidence: 99%

“…We then tested whether ACE can also distinguish subsets of endothelial cells in vitro and track hematopoietic potential in hPSC hematopoietic cultures. For this, we differentiated the human embryonic stem cell (hESC) line H1 using our protocol that specifies hPSCs into WNT-dependent (WNTd) NOTCH-dependent multipotent HOXA + HECs, indicative of intra-embryonic AGM-like hematopoiesis 27,[30][31][32] . In this setting, we observed that ACE is expressed by virtually all day 8 CD34 + cells, including 90±1% of CD34 + CD43 neg CD73 neg CD184 neg cells that comprise HECs in day 8 WNTd hPSC hematopoietic cultures (Extended data Fig.…”

Section: Ace Expression Distinguishes Endothelial Cells With Distinct...mentioning

confidence: 99%

CD32 allows capturing blood cells emergence in slow motion during human embryonic development

Scarfò

Alezz

Khoury

et al. 2023

Preprint

Self Cite

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During development, in the embryo proper blood cells emerge from a subset of specialized endothelial cells, named hemogenic endothelial cells (HECs), via a process known as endothelial-to-hematopoietic transition (EHT) driven by time-specific Notch signaling activation1. HECs represent an elusive cell population as they are rare and transient, rapidly generating blood cells, and specific markers are lacking. Therefore, it remains unclear how and when the hematopoietic fate is specified and how blood cell emergence is molecularly regulated. Notably, thorough characterization of this process is essential to guide the generation of therapeutic blood products in vitro from human pluripotent stem cells (hPSCs). To identify specific human HEC markers, we performed transcriptomic analysis of 28-32-day human embryos, a developmental stage characterized by active hematopoiesis. We observed that the expression of FCGR2B, encoding for the Fc receptor CD32, is highly enriched in the ACE+CD34+ endothelial cell population that contains HECs. Functional ex vivo analyses confirmed that multilineage hematopoietic potential is highly enriched in CD32+ endothelial cells isolated from human embryos. In addition, clonal analysis revealed that 90% of CD32+ hPSC-derived endothelial cells are bona fide HECs. We leveraged this specificity to study how HECs commit to the blood fate. Remarkably, our analyses indicated that HECs progress through different states culminating with the one identified by CD32 expression. Indeed CD32+ HECs no longer require Notch to generate hematopoietic progeny and display full commitment to hematopoiesis even before the expression of hematopoietic markers. These findings provide a precise method for isolating HECs primed to the blood fate from human embryos and hPSC cultures, thus allowing the efficient generation of hematopoietic cells in vitro.

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Identification of a retinoic acid-dependent haemogenic endothelial progenitor from human pluripotent stem cells

Cited by 22 publications

References 41 publications

IκBα controls dormancy induction in Hematopoietic stem cell development via retinoic acid

IκBα controls dormancy induction in Hematopoietic stem cell development via retinoic acid

Layered immunity and layered leukemogenicity: Developmentally restricted mechanisms of pediatric leukemia initiation

CD32 allows capturing blood cells emergence in slow motion during human embryonic development

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