Regulation of neovasculogenesis in co-cultures of aortic adventitial fibroblasts and microvascular endothelial cells by cell-cell interactions and TGF-β/ALK5 signaling.

Adventitial fibroblasts (AFs) are critical mediators of vascular remodeling. However, the contributions of AFs towards development of vasculature and the specific mechanisms by which these cells regulate physiological expansion of the vasa vasorum, the specialized microvasculature that supplies nutr...

Full description

Bibliographic Details
Main Authors: Rebecca A Scott, Eric W Fowler, Xinqiao Jia, Kristi L Kiick, Robert E Akins
Format: Article
Language:English
Published: Public Library of Science (PLoS) 2020-01-01
Series:PLoS ONE
Online Access:https://doi.org/10.1371/journal.pone.0244243
id doaj-167df628fd6b4103b9bf35cce273d35c
record_format Article
spelling doaj-167df628fd6b4103b9bf35cce273d35c2021-03-12T05:30:57ZengPublic Library of Science (PLoS)PLoS ONE1932-62032020-01-011512e024424310.1371/journal.pone.0244243Regulation of neovasculogenesis in co-cultures of aortic adventitial fibroblasts and microvascular endothelial cells by cell-cell interactions and TGF-β/ALK5 signaling.Rebecca A ScottEric W FowlerXinqiao JiaKristi L KiickRobert E AkinsAdventitial fibroblasts (AFs) are critical mediators of vascular remodeling. However, the contributions of AFs towards development of vasculature and the specific mechanisms by which these cells regulate physiological expansion of the vasa vasorum, the specialized microvasculature that supplies nutrients to the vascular wall, are not well understood. To determine the regulatory role of AFs in microvascular endothelial cell (MVEC) neovasculogenesis and to investigate the regulatory pathways utilized for communication between the two cell types, AFs and MVECs were cultured together in poly(ethylene glycol)-based hydrogels. Following preliminary evaluation of a set of cell adhesion peptides (AG10, AG73, A2G78, YIGSR, RGD), 7.5wt% hydrogels containing 3 mM RGD were selected as these substrates did not initiate primitive tubule structures in 3D MVEC monocultures, thus providing a passive platform to study AF-MVEC interaction. The addition of AFs to hydrogels promoted MVEC viability; however, increasing AF density within hydrogels stimulated MVEC proliferation, increased microvessel density and size, and enhanced deposition of basement membrane proteins, collagen IV and laminin. Importantly, AF-MVEC communication through the transforming growth factor beta (TGF-β)/activin receptor-like kinase 5 (ALK5) signaling pathway was observed to mediate microvessel formation, as inhibition of ALK5 significantly decreased MVEC proliferation, microvessel formation, mural cell recruitment, and basement membrane production. These data indicate that AFs regulate MVEC neovasculogenesis and suggest that therapeutics targeting the TGF-β/ALK5 pathway may be useful for regulation of vasculogenic and anti-vasculogenic responses.https://doi.org/10.1371/journal.pone.0244243
collection DOAJ
language English
format Article
sources DOAJ
author Rebecca A Scott
Eric W Fowler
Xinqiao Jia
Kristi L Kiick
Robert E Akins
spellingShingle Rebecca A Scott
Eric W Fowler
Xinqiao Jia
Kristi L Kiick
Robert E Akins
Regulation of neovasculogenesis in co-cultures of aortic adventitial fibroblasts and microvascular endothelial cells by cell-cell interactions and TGF-β/ALK5 signaling.
PLoS ONE
author_facet Rebecca A Scott
Eric W Fowler
Xinqiao Jia
Kristi L Kiick
Robert E Akins
author_sort Rebecca A Scott
title Regulation of neovasculogenesis in co-cultures of aortic adventitial fibroblasts and microvascular endothelial cells by cell-cell interactions and TGF-β/ALK5 signaling.
title_short Regulation of neovasculogenesis in co-cultures of aortic adventitial fibroblasts and microvascular endothelial cells by cell-cell interactions and TGF-β/ALK5 signaling.
title_full Regulation of neovasculogenesis in co-cultures of aortic adventitial fibroblasts and microvascular endothelial cells by cell-cell interactions and TGF-β/ALK5 signaling.
title_fullStr Regulation of neovasculogenesis in co-cultures of aortic adventitial fibroblasts and microvascular endothelial cells by cell-cell interactions and TGF-β/ALK5 signaling.
title_full_unstemmed Regulation of neovasculogenesis in co-cultures of aortic adventitial fibroblasts and microvascular endothelial cells by cell-cell interactions and TGF-β/ALK5 signaling.
title_sort regulation of neovasculogenesis in co-cultures of aortic adventitial fibroblasts and microvascular endothelial cells by cell-cell interactions and tgf-β/alk5 signaling.
publisher Public Library of Science (PLoS)
series PLoS ONE
issn 1932-6203
publishDate 2020-01-01
description Adventitial fibroblasts (AFs) are critical mediators of vascular remodeling. However, the contributions of AFs towards development of vasculature and the specific mechanisms by which these cells regulate physiological expansion of the vasa vasorum, the specialized microvasculature that supplies nutrients to the vascular wall, are not well understood. To determine the regulatory role of AFs in microvascular endothelial cell (MVEC) neovasculogenesis and to investigate the regulatory pathways utilized for communication between the two cell types, AFs and MVECs were cultured together in poly(ethylene glycol)-based hydrogels. Following preliminary evaluation of a set of cell adhesion peptides (AG10, AG73, A2G78, YIGSR, RGD), 7.5wt% hydrogels containing 3 mM RGD were selected as these substrates did not initiate primitive tubule structures in 3D MVEC monocultures, thus providing a passive platform to study AF-MVEC interaction. The addition of AFs to hydrogels promoted MVEC viability; however, increasing AF density within hydrogels stimulated MVEC proliferation, increased microvessel density and size, and enhanced deposition of basement membrane proteins, collagen IV and laminin. Importantly, AF-MVEC communication through the transforming growth factor beta (TGF-β)/activin receptor-like kinase 5 (ALK5) signaling pathway was observed to mediate microvessel formation, as inhibition of ALK5 significantly decreased MVEC proliferation, microvessel formation, mural cell recruitment, and basement membrane production. These data indicate that AFs regulate MVEC neovasculogenesis and suggest that therapeutics targeting the TGF-β/ALK5 pathway may be useful for regulation of vasculogenic and anti-vasculogenic responses.
url https://doi.org/10.1371/journal.pone.0244243
work_keys_str_mv AT rebeccaascott regulationofneovasculogenesisincoculturesofaorticadventitialfibroblastsandmicrovascularendothelialcellsbycellcellinteractionsandtgfbalk5signaling
AT ericwfowler regulationofneovasculogenesisincoculturesofaorticadventitialfibroblastsandmicrovascularendothelialcellsbycellcellinteractionsandtgfbalk5signaling
AT xinqiaojia regulationofneovasculogenesisincoculturesofaorticadventitialfibroblastsandmicrovascularendothelialcellsbycellcellinteractionsandtgfbalk5signaling
AT kristilkiick regulationofneovasculogenesisincoculturesofaorticadventitialfibroblastsandmicrovascularendothelialcellsbycellcellinteractionsandtgfbalk5signaling
AT roberteakins regulationofneovasculogenesisincoculturesofaorticadventitialfibroblastsandmicrovascularendothelialcellsbycellcellinteractionsandtgfbalk5signaling
_version_ 1714787480644878336