Expression of Podocalyxin Separates the Hematopoietic and Vascular Potentials of Mouse Embryonic Stem Cell-Derived Mesoderm

Zhang, Hailan; Nieves, Johnathan L.; Fraser, Stuart T.; Isern, Joan; Douvaras, Panagiotis; Papatsenko, Dmitri; D’Souza, Sunita L.; Lemischka, Ihor R.; Dyer, Michael A.; Baron, Margaret H.

doi:10.1002/stem.1536

Cited by 15 publications

(14 citation statements)

References 54 publications

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“…Additional studies have identified markers to differentiate between hemangioblast and hemogenic endothelium, such as cell-surface intercellular adhesion molecule 2 (ICAM2) (Pearson et al, 2010) and the Ets-related protein encoding transcription factor ETV2 (Wareing et al, 2012). Other molecules such as podocalyxin (Zhang et al, 2014) have been shown to distinguish primitive from definitive hematopoietic potentials.…”

Section: The Next Step: How Observations In Yolk Sac Hematopoiesis Camentioning

confidence: 99%

Modeling murine yolk sac hematopoiesis with embryonic stem cell culture systems

Cook

2014

Front. Biol.

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The onset of hematopoiesis in mammals is defined by generation of primitive erythrocytes and macrophage progenitors in embryonic yolk sac. Laboratories have met the challenge of transient and swiftly changing specification events from ventral mesoderm through multipotent progenitors and maturing lineage-restricted hematopoietic subtypes, by developing powerful in vitro experimental models to interrogate hematopoietic ontogeny. Most importantly, studies of differentiating embryonic stem cell derivatives in embryoid body and stromal coculture systems have identified crucial roles for transcription factor networks (e.g. Gata1, Runx1, Scl) and signaling pathways (e.g. BMP, VEGF, WNT) in controlling stem and progenitor cell output. These and other relevant pathways have pleiotropic biological effects, and are often associated with early embryonic lethality in knockout mice. Further refinement in subsequent studies has allowed conditional expression of key regulatory genes, and isolation of progenitors via cell surface markers (e.g. FLK1) and reporter-tagged constructs, with the purpose of measuring their primitive and definitive hematopoietic potential. These observations continue to inform attempts to direct the differentiation, and augment the expansion, of progenitors in human cell culture systems that may prove useful in cell replacement therapies for hematopoietic deficiencies. The purpose of this review is to survey the extant literature on the use of differentiating murine embryonic stem cells in culture to model the developmental process of yolk sac hematopoiesis.

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Section: The Next Step: How Observations In Yolk Sac Hematopoiesis Camentioning

confidence: 99%

Modeling murine yolk sac hematopoiesis with embryonic stem cell culture systems

Cook

2014

Front. Biol.

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Section: Discussionmentioning

confidence: 99%

“…Zhang et al [30] showed that podocalyxin (podxl) expression separates the hematopoietic and cardiac potential of mouse ESC‐derived Flk1 + mesoderm. Flk1 + cells expressing podxl showed definitive hematopoietic potential, and Flk1 + cells lacking podxl gave rise to cardiomyocytes [30]. Their results, combined with our data, would indicate that the characteristics of F + podxl + cells and F + podxl − cells resemble those of F + C − cells and F + C + cells, respectively.…”

Section: Discussionmentioning

confidence: 99%

Expression of Coxsackievirus and Adenovirus Receptor Separates Hematopoietic and Cardiac Progenitor Cells in Fetal Liver Kinase 1-Expressing Mesoderm

Tashiro

Hirata

Okada

et al. 2015

Stem Cells Translational Medicine

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In developing embryos or in vitro differentiation cultures using pluripotent stem cells (PSCs), such as embryonic stem cells and induced pluripotent stem cells, fetal liver kinase 1 (Flk1)‐expressing mesodermal cells are thought to be a heterogeneous population that includes hematopoietic progenitors, endothelial progenitors, and cardiac progenitors. However, information on cell surface markers for separating these progenitors in Flk1+ cells is currently limited. In the present study, we show that distinct types of progenitor cells in Flk1+ cells could be separated according to the expression of coxsackievirus and adenovirus receptor (CAR, also known as CXADR), a tight junction component molecule. We found that mouse and human PSC‐ and mouse embryo‐derived Flk1+ cells could be subdivided into Flk1+CAR+ cells and Flk1+CAR− cells. The progenitor cells with cardiac potential were almost entirely restricted to Flk1+CAR+ cells, and Flk1+CAR− cells efficiently differentiated into hematopoietic cells. Endothelial differentiation potential was observed in both populations. Furthermore, from the expression of CAR, Flk1, and platelet‐derived growth factor receptor‐α (PDGFRα), Flk1+ cells could be separated into three populations (Flk1+PDGFRα−CAR− cells, Flk1+PDGFRα−CAR+ cells, and Flk1+PDGFRα+CAR+ cells). Flk1+PDGFRα+ cells and Flk1+PDGFRα− cells have been reported as cardiac and hematopoietic progenitor cells, respectively. We identified a novel population (Flk1+PDGFRα−CAR+ cells) with the potential to differentiate into not only hematopoietic cells and endothelial cells but also cardiomyocytes. Our findings indicate that CAR would be a novel and prominent marker for separating PSC‐ and embryo‐derived Flk1+ mesodermal cells with distinct differentiation potentials.

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“…Investigations of Podxl’s actions have predominantly focused on renal podocytes, in which Podxl is important for podocyte foot extension and glomerular diaphragm slit formation [3]. Podxl has also been implicated in supporting vascular integrity [4], and its co-expression with Flk1 distinguishes embryonic Flk1 pos Podxl neg endothelial precursor mesodermal cells from definitive Flk1 pos Podxl pos hematopoietic progenitors [5]. Podxl additionally can mark sub-populations of embryonic CD34 pos hematopoietic stem cells [6], and Podxl/Pclp1 has been defined as functional marker of hematopoiesis within AGM regions [7].…”

Section: Introductionmentioning

confidence: 99%

Novel roles for podocalyxin in regulating stress myelopoiesis, Rap1a, and neutrophil migration

Karaczyn

McGlauflin

et al. 2017

Experimental Hematology

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Podocalyxin (Podxl) is a CD34 orthologue and cell surface sialomucin with reported roles in renal podocyte diaphragm slit development, vascular cell integrity, and the progression of blood, breast, and prostate cancers. Roles for Podxl during non-malignant hematopoiesis, however, are largely undefined. Presently we have developed a Vav-Cre Podxl knockout mouse model, and report on novel roles for Podxl in governing stress myelopoiesis. At steady-state, Podxl expression among hematopoietic progenitor cells was low-level but was induced by GCSF (granulocyte colony stimulating factor) in myeloid progenitors, and by TPO (thrombopoietin) in HSCs. In keeping with low level Podxl expression at steady-state, Vav-Cre deletion of Podxl did not markedly alter peripheral blood cell levels. G-CSF challenge in Podxl-KO mice, in contrast, hyper-elevated peripheral blood neutrophil and monocyte levels. Podxl-KO also substantially heightened neutrophil levels following 5-fluorouracil myeloablation. These LOF phenotypes were selective, and Podxl-KO did not alter lymphocyte, basophil or eosinophil levels. Within bone marrow (and following G-CSF challenge), Podxl deletion moderately decreased CFU-GEMM and CD16/32posCD11bpos progenitors but did not affect Gr-1pos cell populations. Notably, Podxl-KO did significantly heighten peripheral blood neutrophil migration capacities. To interrogate Podxl’s action mechanisms, a co-immunoprecipitation plus LC-MS/MS (liquid chromatography – mass spectrometry) approach was applied using hematopoietic progenitors from G-CSF-challenged mice. Rap1a, a Ras-related small GTPase, was a predominant co-retrieved Podxl partner. In bone marrow HPC’s, Podxl-KO led to heightened GCSF activation of Rap1aGTP, and Rap1aGTP inhibition attenuated Podxl-KO neutrophil migration. Studies reveal novel roles for Podxl as an important modulator of neutrophil and monocyte formation, and of Rap1a activation, during stress hematopoiesis.

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Expression of Podocalyxin Separates the Hematopoietic and Vascular Potentials of Mouse Embryonic Stem Cell-Derived Mesoderm

Cited by 15 publications

References 54 publications

Modeling murine yolk sac hematopoiesis with embryonic stem cell culture systems

Modeling murine yolk sac hematopoiesis with embryonic stem cell culture systems

Expression of Coxsackievirus and Adenovirus Receptor Separates Hematopoietic and Cardiac Progenitor Cells in Fetal Liver Kinase 1-Expressing Mesoderm

Novel roles for podocalyxin in regulating stress myelopoiesis, Rap1a, and neutrophil migration

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