Reversible transdifferentiation of blood vascular endothelial cells to a lymphatic-like phenotype in vitro

Cooley, Lindsay S.; Handsley, Madeleine M.; Zhou, Zhigang; Lafleur, Michel; Pennington, Caroline J.; Thompson, Erik W.; Pöschl, Ernst; Edwards, Dylan R.

doi:10.1242/jcs.064279

Cited by 44 publications

(28 citation statements)

References 39 publications

(21 reference statements)

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“…However, upon assessment of EC potential after serial passages, we observed that sorted and matured ECs lost some VEcad expression by passage 4, warranting future work that will examine means to retain endothelial potential over numerous passages via media or culture optimization. These findings are in agreement with a previous study, which also demonstrated loss of vascular EC marker expression from mature, fully differentiated ECs (i.e., human umbilical vein ECs) and their subsequent transdifferentiation toward lymphatic ECs in vitro [32]. Moreover, previous studies report that PSCderived ECs lose their endothelial potential over serial passaging, perhaps owing to the notion that these derivatives may still exhibit some plasticity [33].…”

Section: Discussionsupporting

confidence: 93%

Characterizing Human Pluripotent-Stem-Cell-Derived Vascular Cells for Tissue Engineering Applications

Kusuma

Facklam

Gerecht

2015

Stem Cells and Development

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Tissue-engineered constructs are rendered useless without a functional vasculature owing to a lack of nutrients and oxygen. Cell-based approaches to reconstruct blood vessels can yield structures that mimic native vasculature and aid transplantation. Vascular derivatives of human induced pluripotent stem cells (hiPSCs) offer opportunities to generate patient-specific therapies and potentially provide unlimited amounts of vascular cells. To be used in engineered vascular constructs and confer therapeutic benefit, vascular derivatives must exhibit additional key properties, including extracellular matrix (ECM) production to confer structural integrity and growth factor production to facilitate integration. In this study, we examine the hypothesis that vascular cells derived from hiPSCs exhibit these critical properties to facilitate their use in engineered tissues. hiPSCs were codifferentiated toward early vascular cells (EVCs), a bicellular population of endothelial cells (ECs) and pericytes, under varying low-oxygen differentiation conditions; subsequently, ECs were isolated and passaged. We found that EVCs differentiated under low-oxygen conditions produced copious amounts of collagen IV and fibronectin as well as vascular endothelial growth factor and angiopoietin 2. EVCs differentiated under atmospheric conditions did not demonstrate such abundant ECM expression, but exhibited greater expression of angiopoietin 1. Isolated ECs could proliferate up to three passages while maintaining the EC marker vascular endothelial cadherin. Isolated ECs demonstrated an increased propensity to produce ECM compared with their EVC correlates and took on an arterial-like fate. These findings illustrate that hiPSC vascular derivates hold great potential for therapeutic use and should continue to be a preferred cell source for vascular construction.

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Section: Discussionsupporting

confidence: 93%

Characterizing Human Pluripotent-Stem-Cell-Derived Vascular Cells for Tissue Engineering Applications

Kusuma

Facklam

Gerecht

2015

Stem Cells and Development

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“…Interestingly, a recent lineage-tracing analysis shows that PDGFRβ-positive endothelium contributes to the CLV formation during embryonal development [14], implying that upon severe tissue injury, blood endothelial cells acquire the lymphatic endothelial cell phenotype. Such endothelial cell plasticity has been previously described in cultured cells and for the tumor vasculature [43–45]. …”

Section: Resultsmentioning

confidence: 78%

Phenotypically heterogeneous podoplanin-expressing cell populations are associated with the lymphatic vessel growth and fibrogenic responses in the acutely and chronically infarcted myocardium

et al. 2017

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Cardiac lymphatic vasculature undergoes substantial expansion in response to myocardial infarction (MI). However, there is limited information on the cellular mechanisms mediating post-MI lymphangiogenesis and accompanying fibrosis in the infarcted adult heart. Using a mouse model of permanent coronary artery ligation, we examined spatiotemporal changes in the expression of lymphendothelial and mesenchymal markers in the acutely and chronically infarcted myocardium. We found that at the time of wound granulation, a three-fold increase in the frequency of podoplanin-labeled cells occurred in the infarcted hearts compared to non-operated and sham-operated counterparts. Podoplanin immunoreactivity detected LYVE-1-positive lymphatic vessels, as well as masses of LYVE-1-negative cells dispersed between myocytes, predominantly in the vicinity of the infarcted region. Podoplanin-carrying populations displayed a mesenchymal progenitor marker PDGFRα, and intermittently expressed Prox-1, a master regulator of the lymphatic endothelial fate. At the stages of scar formation and maturation, concomitantly with the enlargement of lymphatic network in the injured myocardium, the podoplanin-rich LYVE-1-negative multicellular assemblies were apparent in the fibrotic area, aligned with extracellular matrix deposits, or located in immediate proximity to activated blood vessels with high VEGFR-2 content. Of note, these podoplanin-containing cells acquired the expression of PDGFRβ or a hematoendothelial epitope CD34. Although Prox-1 labeling was abundant in the area affected by MI, the podoplanin-presenting cells were not consistently Prox-1-positive. The concordance of podoplanin with VEGFR-3 similarly varied. Thus, our data reveal previously unknown phenotypic and structural heterogeneity within the podoplanin-positive cell compartment in the infarcted heart, and suggest an alternate ability of podoplanin-presenting cardiac cells to generate lymphatic endothelium and pro-fibrotic cells, contributing to scar development.

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“…Endothelial growth medium-EGM2 (C22011; PromoCell, Birmingham, UK) with supplements was used as culture medium for HuVECs according to the manufacturer's protocols. Murine perivascular cells (PVC) were isolated as previously described and used between passages 32 and 38 (35). Pericytes were routinely incubated in Dulbecco's modified Eagle's medium (DMEM) containing 10% fetal bovine serum (FBS).…”

Section: Methodsmentioning

confidence: 99%

Differential effects of sulforaphane in regulation of angiogenesis in a co-culture model of endothelial cells and pericytes

et al. 2017

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Abstract. Aberrant neovascularization supports nutrients and the oxygen microenvironment in tumour growth, invasion and metastasis. Recapitulation of functional microvascular structures in vitro could provide a platform for the study of vascular conditions. Sulforaphane (SFN), an isothiocyanate, has been reported to possess chemopreventive properties. In the present study, the effects of SFN on cell proliferation and tubular formation have been investigated using endothelial cells (ECs) and pericytes in coculture. SFN showed a dosedependent inhibition on the growth of ECs and pericytes with IC 50 values 46.7 and 32.4 µM, respectively. SFN (5-20 µM) inhibited tube formation in a 3D coculture although a lower dose (1.25 µM) promoted 30% more endothelial tube formation than control. Moreover, SFN affected intercellular communication between ECs and pericytes via inhibition of angiogenic factor such as vascular endothelial growth factor (VEGF) expression in pericytes. However, the expression of its receptor (VEGFR-2) was found significantly increased in ECs. These effects were associated with downregulation of prolyl hydroxylase domain-containing protein 1 and 2 (PHD1/2) and activation of hypoxia-inducible factor-1 (HIF) pathway by SFN. Furthermore, thioredoxin reductase-1 was also upregulated by SFN treatment, suggesting that antioxidant and redox regulation are involved in angiogenesis. Taken together, the results of the present study suggest that SFN differentially regulates endothelial cells and pericytes disrupting their interplay through the VEGF-VEGFR signalling pathway. Anti-angiogenesis property of SFN indicates that it has potential role as an anticancer agent. IntroductionAngiogenesis, the growth of new capillary blood vessels, is a normal and vital process in growth, development and wound healing. However, it is also a fundamental step in the growth of tumours. Nutrients and oxygen, supplied by the blood vessels into the tumours, are essential for the growth and progression of malignant tumours beyond the size of 1-2 mm 3 (1). Newly formed blood vessels can facilitate cells escaping through leakage from primary tumour sites to metastasize, the major cause of mortality for cancer patients. Anti-angiogenesis has been recognized as valuable therapy in treatment of various metastatic cancers since the theory was first proposed by Folkman (2,3). Tumour angiogenesis is believed to be regulated by the interactions between pro-angiogenic and anti-angiogenic factors in the tumour microenvironment (4). Angiogenic response is a dynamic process requiring a series of fine-tuned angiogenic signalling and molecular events. Hypoxia is the common inducer of angiogenesis in the core of large tumours stimulating the release of pro-angiogenic factors to promote endothelial cell proliferation, migration, differentiation and self-assembly into vascular-like structures. Subsequently, perivascular cells are recruited to form mature and stable vessels (5).The interaction between endothelial cells (ECs) and pericytes (PVCs) has g...

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Reversible transdifferentiation of blood vascular endothelial cells to a lymphatic-like phenotype in vitro

Cited by 44 publications

References 39 publications

Characterizing Human Pluripotent-Stem-Cell-Derived Vascular Cells for Tissue Engineering Applications

Characterizing Human Pluripotent-Stem-Cell-Derived Vascular Cells for Tissue Engineering Applications

Phenotypically heterogeneous podoplanin-expressing cell populations are associated with the lymphatic vessel growth and fibrogenic responses in the acutely and chronically infarcted myocardium

Differential effects of sulforaphane in regulation of angiogenesis in a co-culture model of endothelial cells and pericytes

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