Inducible Gata1 suppression expands megakaryocyte-erythroid progenitors from embryonic stem cells

Noh, Jin-Yong; Gandre-Babbe, Shilpa; Wang, Yuhuan; Hayes, Vincent; Yao, Yu; Gadue, Paul; Sullivan, Spencer K.; Chou, Stella T.; Machlus, Kellie R.; Italiano, Joseph E.; Kyba, Michael; Finkelstein, David; Ulirsch, Jacob C.; Sankaran, Vijay G.; French, Deborah L.; Poncz, Mortimer; Weiss, Mitchell J.

doi:10.1172/jci77670

Cited by 33 publications

(31 citation statements)

References 36 publications

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“…An improved second-generation system was created, and these cells are designated G1ME2. 57 This system functions under more physiologic conditions using a dox-inducible promoter for knocking down expression of GATA1 in mouse ESC-derived MEPs. In the presence of dox and GATA1 repression, G1ME2 cells expanded logarithmically for .40 days.…”

Section: Generating Megakaryocyte Progenitor Cellsmentioning

confidence: 99%

“…(A) Self-replicating human PSCs can be genetically manipulated using genome-editing technologies. 67 41,57 (C-D) Megakaryocyte maturation and PPF may be induced using different matrices, modulators of apoptosis, and shear stress. 65,66 Platelet release could also be induced from mature megakaryocytes by inflammatory induced rupture of the cell.…”

Section: Authorshipmentioning

confidence: 99%

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Understanding platelet generation from megakaryocytes: implications for in vitro–derived platelets

Sim

Poncz

Gadue

et al. 2016

Blood

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101

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Platelets are anucleate cytoplasmic discs derived from megakaryocytes that circulate in the blood and have major roles in hemostasis, thrombosis, inflammation, and vascular biology. Platelet transfusions are required to prevent the potentially life-threatening complications of severe thrombocytopenia seen in a variety of medical settings including cancer therapy, trauma, and sepsis. Platelets used in the clinic are currently donor-derived which is associated with concerns over sufficient availability, quality, and complications due to immunologic and/or infectious issues. To overcome our dependence on donor-derived platelets for transfusion, efforts have been made to generate in vitro–based platelets. Work in this area has advanced our understanding of the complex processes that megakaryocytes must undergo to generate platelets both in vivo and in vitro. This knowledge has also defined the challenges that must be overcome to bring in vitro–based platelet manufacturing to a clinical reality. This review will focus on our understanding of committed megakaryocytes and platelet release in vivo and in vitro, and how this knowledge can guide the development of in vitro–derived platelets for clinical application.

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Section: Generating Megakaryocyte Progenitor Cellsmentioning

confidence: 99%

Section: Authorshipmentioning

confidence: 99%

Understanding platelet generation from megakaryocytes: implications for in vitro–derived platelets

Sim

Poncz

Gadue

et al. 2016

Blood

Self Cite

101

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“…Furthermore, an arteriole laser injury model revealed that these platelets are incorporated into functional thrombi at a rate that is indistinguishable from the rates of incorporation of either platelets produced from fetal liver-derived megakaryocytes or platelets isolated from donor mice. These experiments provide in vivo evidence that G1ME2-derived megakaryocytes can produce platelets able to home to sites of injury and participate in thrombus formation (20).…”

Section: Thrombocytopenia and The Need For Alternative Platelet Sourcesmentioning

confidence: 91%

“…In the presence of doxycycline, which induces shRNA-mediated GATA1 suppression, these ex vivo-differentiated mES cells exhibited logarithmic growth for over 40 days as MEP-like cells (referred to as G1ME2 cells), resulting in an approximately 10 13 -fold expansion in cell number. Upon doxycycline withdrawal, which restores GATA1, approximately 20% of G1ME2 cells differentiated into cells with morphological and expression patterns that were consistent with mature megakaryocytes (20). Furthermore, these G1ME2-derived megakaryocytes generated proplatelet processes, a hallmark of platelet generation (Figure 1).…”

Section: Thrombocytopenia and The Need For Alternative Platelet Sourcesmentioning

confidence: 99%

Embryonic stem cells as sources of donor-independent platelets

Canver¹,

Bauer²,

Orkin³

2015

J. Clin. Invest.

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C o m m e n t a r y Thrombocytopenia and the need for alternative platelet sourcesThe blood system comprises a hierarchy of rare, largely quiescent, self-renewing hematopoietic stem cells (HSCs) and highly proliferative lineage-committed progenitors and precursors in the bone marrow, which yield a continual supply of mature blood cells of diverse lineages (1). The nearly one trillion platelets present in circulation derive from bone marrow-resident, multinucleated megakaryocytes, which in turn derive from bipotential megakaryocyte-erythroid progenitors (MEPs). The master transcription factor GATA1 is required for maturation of MEPs into both megakaryocytic and erythroid lineages (1, 2). It is estimated that each megakaryocyte produces approximately 10 3 to 10 4 platelets, which have half-lives of only about three days in the circulation (3-6). Reduction in the number of circulating platelets, termed thrombocytopenia, is a common clinical condition with a variety of etiologies, including infectious and heritable disease, malignancy, liver failure, chemotherapy, and medication side effects (1, 4, 7-9).Transfusion of donor-derived platelets is employed with increasing frequency to treat patients with thrombocytopenia and places a high demand on platelet donation (10). Over two million platelet units are transfused annually in the United States (8, 10); however, there are multiple factors that hinder platelet transfusion. Donor platelets have only a three-to seven-day shelf life, limiting the ability to store and transport these cells and generating substantial waste through the disposal of aged platelets. Intermittent shortages arise due to a mismatch between supply and demand (11,12). Further, donor-derived platelets carry the inherent risk of viral transmission and bacterial contamination due to extended room temperature storage (8). Together, these limitations have motivated the development of sustainable donor-independent platelet sources to meet the growing need for these cells (7,11).Pluripotent embryonic stem (ES) cells or induced pluripotent stem (iPS) cells have been proposed as a potential donor-independent source of platelets. One vision is that either ES cells or iPS cells could be used to produce a renewable and immortalized progenitor-like cell line that retains the capacity to differentiate into mature megakaryocytes, which in turn would produce functional platelets. To date, attempts to employ this strategy have suffered from a poor output of PS cell-derived megakaryocytes. Moreover, inefficient generation of platelets from megakaryocytes in vitro has been an additional challenge (4,6,11,13,14). The GATA switch and regulation of megakaryopoiesisThe study of the GATA transcription factors GATA1 and GATA2 has provided insight into normal and disease-associated megakaryopoiesis. While GATA2 is highly expressed in early stem and progenitor cells, megakaryopoiesis and erythropoiesis are accompanied by a concurrent downregulation of GATA2 and upregulation of the closely related GATA1, a process referred t...

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“…4 It is not clear whether preconditioning affects the potential for an immune response or immune tolerance in the context of platelet-derived FVIII. Furthermore, if current efforts [8][9][10][11][12][13] to generate platelets or megakaryocytes in vitro succeed, genetically manipulated platelets containing FVIII may be used therapeutically as a potential transfusion alternative, in HA patients even with inhibitors. One important concern that has not been explored, however, is the immunogenicity of plateletderived FVIII.…”

Section: Introductionmentioning

confidence: 99%

The immunogenicity of platelet-derived FVIII in hemophilia A mice with or without preexisting anti-FVIII immunity

Chen

Schroeder

Chen

et al. 2016

Blood

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Inducible Gata1 suppression expands megakaryocyte-erythroid progenitors from embryonic stem cells

Cited by 33 publications

References 36 publications

Understanding platelet generation from megakaryocytes: implications for in vitro–derived platelets

Understanding platelet generation from megakaryocytes: implications for in vitro–derived platelets

Embryonic stem cells as sources of donor-independent platelets

The immunogenicity of platelet-derived FVIII in hemophilia A mice with or without preexisting anti-FVIII immunity

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