The requirement for fibroblasts in angiogenesis: fibroblast-derived matrix proteins are essential for endothelial cell lumen formation

Newman, Andrew; Nakatsu, Martin N.; Chou, Wayne; Gershon, Paul D.; Hughes, Christopher C.W.

doi:10.1091/mbc.e11-05-0393

Cited by 398 publications

(405 citation statements)

References 57 publications

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“…Interestingly, our results suggest that different combinations of factors can be similarly potent. In line with these findings, one study reported a combination of factors secreted by stromal fibroblasts that induced sprouting (39), and another found a combination of hematopoietic chemokines led to a marked enhancement in tubulogenesis and sprouting (40). The recognition that multiple combinations of factors can drive angiogenesis, likely through different mechanisms, further underscores an important role for model systems that allow for the rapid characterization of factor combinations.…”

Section: Discussionmentioning

confidence: 76%

Biomimetic model to reconstitute angiogenic sprouting morphogenesis in vitro

Nguyen

Stapleton²,

Yang

et al. 2013

Proc. Natl. Acad. Sci. U.S.A.

428

441

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Angiogenesis is a complex morphogenetic process whereby endothelial cells from existing vessels invade as multicellular sprouts to form new vessels. Here, we have engineered a unique organotypic model of angiogenic sprouting and neovessel formation that originates from preformed artificial vessels fully encapsulated within a 3D extracellular matrix. Using this model, we screened the effects of angiogenic factors and identified two distinct cocktails that promoted robust multicellular endothelial sprouting. The angiogenic sprouts in our system exhibited hallmark structural features of in vivo angiogenesis, including directed invasion of leading cells that developed filopodia-like protrusions characteristic of tip cells, following stalk cells exhibiting apical-basal polarity, and lumens and branches connecting back to the parent vessels. Ultimately, sprouts bridged between preformed channels and formed perfusable neovessels. Using this model, we investigated the effects of angiogenic inhibitors on sprouting morphogenesis. Interestingly, the ability of VEGF receptor 2 inhibition to antagonize filopodia formation in tip cells was context-dependent, suggesting a mechanism by which vessels might be able to toggle between VEGF-dependent and VEGFindependent modes of angiogenesis. Like VEGF, sphingosine-1-phosphate also seemed to exert its proangiogenic effects by stimulating directional filopodial extension, whereas matrix metalloproteinase inhibitors prevented sprout extension but had no impact on filopodial formation. Together, these results demonstrate an in vitro 3D biomimetic model that reconstitutes the morphogenetic steps of angiogenic sprouting and highlight the potential utility of the model to elucidate the molecular mechanisms that coordinate the complex series of events involved in neovascularization.A ngiogenesis, the process by which new capillary vessels sprout from existing vasculature, plays a critical role in embryonic development and wound healing, and its dysregulation can contribute to cancer progression as well as numerous inflammatory and ischemic diseases (1, 2). Consequently, therapeutic strategies to suppress, enhance, or normalize angiogenesis are widely sought to treat a broad spectrum of diseases (1, 2). The most mature among these approaches targets the activity of angiogenic growth factors, such as vascular endothelial growth factor (VEGF), to modulate relevant signaling pathways and control the angiogenesis process. Indeed, inhibitors of such pathways have emerged as a mainstay therapy for some cancers and diabetic retinopathy (3-5). However, it is still unclear how the endothelial cells (ECs) lining blood vessels form new vessels, or how angiogenic factors regulate such a dynamic, multicellular process.Examining the physical process of angiogenesis requires experimental systems in which the formation of new capillary vessels can be easily observed and manipulated. Commonly used in vivo models such as the mouse dorsal window chamber, chick chorioallantoic membrane, and mouse corneal micro...

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Section: Discussionmentioning

confidence: 76%

Biomimetic model to reconstitute angiogenic sprouting morphogenesis in vitro

Nguyen

Stapleton²,

Yang

et al. 2013

Proc. Natl. Acad. Sci. U.S.A.

428

441

View full text Add to dashboard Cite

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“…A third possibility is that the various stromal cell types secrete distinct pro-and antiangiogenic cytokines that regulate vessel maturation, and/or differentially contribute to the production of new ECM proteins required for vessel formation and stability. 47 A final possibility relates to the influence of stromal cell identity on the proteolytic remodeling of the ECM during the neovascularization process. 24,48 Our published data suggest that capillary formation induced by fibroblasts is very fast (perhaps tumor-like), resulting from a plasmin-mediated proteolysis that rapidly degrades the matrix and disrupts its mechanical properties in a global fashion.…”

Section: Grainger Et Almentioning

confidence: 99%

Stromal Cell Identity Influences the In Vivo Functionality of Engineered Capillary Networks Formed by Co-delivery of Endothelial Cells and Stromal Cells

Grainger

Carrion

Ceccarelli

et al. 2013

Tissue Engineering Part A

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A major translational challenge in the fields of therapeutic angiogenesis and tissue engineering is the ability to form functional networks of blood vessels. Cell-based strategies to promote neovascularization have been widely explored, and have led to the consensus that co-delivery of endothelial cells (ECs) (or their progenitors) with some sort of a supporting stromal cell type is the most effective approach. However, the choice of stromal cells has varied widely across studies, and their impact on the functional qualities of the capillaries produced has not been examined. In this study, we injected human umbilical vein ECs alone or with normal human lung fibroblasts (NHLFs), human bone marrow-derived mesenchymal stem cells (BMSCs), or human adipose-derived stem cells (AdSCs) in a fibrin matrix into subcutaneous pockets in SCID mice. All conditions yielded new human-derived vessels that inosculated with mouse vasculature and perfused the implant, but there were significant functional differences in the capillary networks, depending heavily on the identity of the co-delivered stromal cells. EC-alone and EC-NHLF implants yielded immature capillary beds characterized by high levels of erythrocyte pooling in the surrounding matrix. EC-BMSC and EC-AdSC implants produced more mature capillaries characterized by less extravascular leakage and the expression of mature pericyte markers. Injection of a fluorescent tracer into the circulation also showed that EC-BMSC and EC-AdSC implants formed vasculature with more tightly regulated permeability. These results suggest that the identity of the stromal cells is key to controlling the functional properties of engineered capillary networks.

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“…24 Understanding this process remains a challenge, since fibroblasts have the capacity to alter the mechanical extracellular microenvironment, thereby regulating vascularization processes. 25 Fibroblast-derived proteins, including growth factors and matrix proteins, have been shown to modulate EC sprouting and the expansion of capillary-like networks in vitro, [26][27][28] contributing to the role of fibroblasts as periendothelial cells in vivo. 29 Thus, the hypothesis underlying herein is that, when cocultured with OECs or mature ECs, different types of fibroblasts will exert distinct influences in the assembly of capillary-like structures.…”

mentioning

confidence: 99%

Fibroblast-Endothelial Partners for Vascularization Strategies in Tissue Engineering

Costa‐Almeida

Gómez-Lázaro

Ramalho

et al. 2015

Tissue Engineering Part A

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Cell-based approaches have emerged as a promising therapy to achieve successful vascularization in tissue engineering. Since fibroblasts activation and migration is required for physiological events relying on angiogenesis, we hypothesize herein that different fibroblasts exhibit distinct capacity to promote capillary-like structures assembly, by mature and progenitor endothelial cells (ECs). Outgrowth endothelial cells (OECs) were isolated from human umbilical cord blood samples and characterized by immunofluorescence and imaging flow cytometry for endothelial markers. Coculture systems were established using either human umbilical vein ECs (HUVECs) or OECs with fibroblasts, being evaluated at 7, 14, and 21 days of culture. Two types of human dermal fibroblasts (HDF) were used, namely neonatal human foreskin fibroblasts-1 (HFF-1) and juvenile HDF. OECs expressed EC markers and formed capillary-like structures. HFF-1 exhibited higher expression of transglutaminase-2, while HDF exhibited a higher expression of a-smooth muscle actin (a-SMA) and podoplanin, which were not observed for HFF-1. Formation of capillary-like structures was only observed in cocultures with HDF and not with HFF-1. No significant differences were found between HDF and OECs or HUVECs cocultures. These findings suggest that HDF is a preferential cell source for promoting vascularization, either using mature or progenitor ECs, probably due to their higher expression of a-SMA and podoplanin, and increased synthesis of extracellular matrix. This work opens new research possibilities regarding the use of specific fibroblast populations cocultured with ECs, as efficient partners for vascular development in regenerative medicine strategies.

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The requirement for fibroblasts in angiogenesis: fibroblast-derived matrix proteins are essential for endothelial cell lumen formation

Cited by 398 publications

References 57 publications

Biomimetic model to reconstitute angiogenic sprouting morphogenesis in vitro

Biomimetic model to reconstitute angiogenic sprouting morphogenesis in vitro

Stromal Cell Identity Influences the In Vivo Functionality of Engineered Capillary Networks Formed by Co-delivery of Endothelial Cells and Stromal Cells

Fibroblast-Endothelial Partners for Vascularization Strategies in Tissue Engineering

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