An image-based screen identifies a small molecule regulator of megakaryopoiesis

Boitano, Anthony E.; Lichtervelde, Lorenzo de; Snead, Jennifer L.; Cooke, M.; Schultz, Peter G.

doi:10.1073/pnas.1212545109

Cited by 16 publications

(14 citation statements)

References 27 publications

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“…[51][52][53][54] Small molecules acting through platelet-derived growth factor receptors or the TPO receptor MPL have been identified as able to induce ex vivo production of MKs and platelets. [55][56][57][58] Our work describes a novel strategy to enhance in vitro MK generation, based on insights from the integrated analysis of a human iPSC model of a monogenic human disease. Our study demonstrated that isogenic iPSC pairs, with or without a specific mutation, not only recapitulate precisely the disease phenotypes but also help discover new druggable targets distinct from the original mutated gene.…”

Section: Discussionmentioning

confidence: 99%

Human NOTCH4 is a key target of RUNX1 in megakaryocytic differentiation

Chen

Bai

et al. 2018

Blood

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Megakaryocytes (MKs) in adult marrow produce platelets that play important roles in blood coagulation and hemostasis. Monoallelic mutations of the master transcription factor gene lead to familial platelet disorder (FPD) characterized by defective MK and platelet development. However, the molecular mechanisms of FPD remain unclear. Previously, we generated human induced pluripotent stem cells (iPSCs) from patients with FPD containing a nonsense mutation. Production of MKs from the FPD-iPSCs was reduced, and targeted correction of the mutation restored MK production. In this study, we used isogenic pairs of FPD-iPSCs and the MK differentiation system to identify RUNX1 target genes. Using integrative genomic analysis of hematopoietic progenitor cells generated from FPD-iPSCs, and mutation-corrected isogenic controls, we identified 2 gene sets the transcription of which is either up- or downregulated by RUNX1 in mutation-corrected iPSCs. Notably, expression was negatively controlled by RUNX1 via a novel regulatory DNA element within the locus, and we examined its involvement in MK generation. Specific inactivation of by an improved CRISPR-Cas9 system in human iPSCs enhanced megakaryopoiesis. Moreover, small molecules known to inhibit Notch signaling promoted MK generation from both normal human iPSCs and postnatal CD34 hematopoietic stem and progenitor cells. Our study newly identified as a RUNX1 target gene and revealed a previously unappreciated role of NOTCH4 signaling in promoting human megakaryopoiesis. Our work suggests that human iPSCs with monogenic mutations have the potential to serve as an invaluable resource for discovery of novel druggable targets.

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Section: Discussionmentioning

confidence: 99%

Human NOTCH4 is a key target of RUNX1 in megakaryocytic differentiation

Chen

Bai

et al. 2018

Blood

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“…Because LOX amplifies PDGF receptor signaling, and because inhibition of this signaling is important for transition of common myeloid progenitors to megakaryocytes, 46 LOX inhibition may be helpful in halting megakaryocyte proliferation in MPNs. Yet, we do not rule out the possibility that LOX influences the MKP pool also via mechanisms independent of PDGF, such as its newly identified influence on collagen receptors.…”

Section: Org Frommentioning

confidence: 99%

Lysyl oxidase is associated with increased thrombosis and platelet reactivity

Matsuura¹,

Mi²,

Koupenova³

et al. 2016

Blood

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• Mice overexpressing LOX in platelets have more severe thrombosis than normal animals.• LOX expression influences platelet adhesion to collagen. ) were generated. Pf4-Lox tg/tg mice had a normal number of platelets; however, time to vessel occlusion after endothelial injury was significantly shorter in Pf4-Lox tg/tg mice, indicating a higher propensity for thrombus formation in vivo.Exploring underlying mechanisms, we found that Pf4-Lox tg/tg platelets adhere better to collagen and have greater aggregation response to lower doses of collagen compared with controls. Platelet activation in response to the ligand for collagen receptor glycoprotein VI (cross-linked collagen-related peptide) was unaffected. However, the higher affinity of Pf4-Lox tg/tg platelets to the collagen sequence GFOGER implies that the collagen receptor integrin a 2 b 1 is affected by LOX. Taken together, our findings demonstrate that LOX enhances platelet activation and thrombosis. (Blood. 2016;127(11):1493-1501

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“…UCB‐derived CD34 cells were expanded in combination with reagents previously reported to increase CD34 cell expansion, as well as on a pseudo‐3D membrane. The aryl hydrocarbon signaling inhibitor SR1, copper chelator TEPA, and caspase inhibitor zVADfmk have been shown to increase the CD34‐fold expansion in vitro. We found that the use of the aryl hydrocarbon receptor inhibitor (SR1) increased the CD34‐fold expansion to a mean of 85‐fold on Day 7 (Fig.…”

Section: Resultsmentioning

confidence: 99%