TY - JOUR
T1 - Extracellular Matrix Analysis of Human Renal Arteries in Both Quiescent and Active Vascular State
AU - van Dijk, Christian G M
AU - Louzao-Martinez, Laura
AU - van Mulligen, Elise
AU - Boermans, Bart
AU - Demmers, Jeroen A A
AU - van den Bosch, Thierry P P
AU - Goumans, Marie-José
AU - Duncker, Dirk J
AU - Verhaar, Marianne C
AU - Cheng, Caroline
N1 - Funding Information:
Funding: This research was funded by The Netherlands Foundation for Cardiovascular Excellence (to C.C.), Netherlands Organization for Scientific Research Vidi grant (no. 91714302 to C.C.), the Erasmus MC fellowship grant (to C.C.), the Regenerative Medicine Fellowship grant of the University Medical Center Utrecht (to C.C.) and The Netherlands Cardiovascular Research Initiative: An initiative with support of the Dutch Heart Foundation (CVON2014-11 RECONNECT to C.C., D.J.D., and M.C.V.).
Funding Information:
This research was funded by The Netherlands Foundation for Cardiovascular Excellence (to C.C.), Netherlands Organization for Scientific Research Vidi grant (no. 91714302 to C.C.), the Erasmus MC fellowship grant(toC.C.),theRegenerativeMedicineFellowshipgrantoftheUniversityMedicalCenterUtrecht(toC.C.)and The Netherlands Cardiovascular Research Initiative: An initiative with support of the Dutch Heart Foundation (CVON2014-11 RECONNECT to C.C., D.J.D., and M.C.V.). Acknowledgments: We would like to thank O.G. de Jong for donating lentiviral GFP constructs and J. Xu for performing (immune) histology staining of the human tissue.
Publisher Copyright:
© 2020 by the authors. . Licensee MDPI, Basel, Switzerland.
PY - 2020/6
Y1 - 2020/6
N2 - In vascular tissue engineering strategies, the addition of vascular-specific extracellular matrix (ECM) components may better mimic the in vivo microenvironment and potentially enhance cell-matrix interactions and subsequent tissue growth. For this purpose, the exact composition of the human vascular ECM first needs to be fully characterized. Most research has focused on characterizing ECM components in mature vascular tissue; however, the developing fetal ECM matches the active environment required in vascular tissue engineering more closely. Consequently, we characterized the ECM protein composition of active (fetal) and quiescent (mature) renal arteries using a proteome analysis of decellularized tissue. The obtained human fetal renal artery ECM proteome dataset contains higher levels of 15 ECM proteins versus the mature renal artery ECM proteome, whereas 16 ECM proteins showed higher levels in the mature tissue compared to fetal. Elastic ECM proteins EMILIN1 and FBN1 are significantly enriched in fetal renal arteries and are mainly produced by cells of mesenchymal origin. We functionally tested the role of EMILIN1 and FBN1 by anchoring the ECM secreted by vascular smooth muscle cells (SMCs) to glass coverslips. This ECM layer was depleted from either EMILIN1 or FBN1 by using siRNA targeting of the SMCs. Cultured endothelial cells (ECs) on this modified ECM layer showed alterations on the transcriptome level of multiple pathways, especially the Rho GTPase controlled pathways. However, no significant alterations in adhesion, migration or proliferation were observed when ECs were cultured on EMILIN1- or FNB1-deficient ECM. To conclude, the proteome analysis identified unique ECM proteins involved in the embryonic development of renal arteries. Alterations in transcriptome levels of ECs cultured on EMILIN1- or FBN1-deficient ECM showed that these candidate proteins could affect the endothelial (regenerative) response.
AB - In vascular tissue engineering strategies, the addition of vascular-specific extracellular matrix (ECM) components may better mimic the in vivo microenvironment and potentially enhance cell-matrix interactions and subsequent tissue growth. For this purpose, the exact composition of the human vascular ECM first needs to be fully characterized. Most research has focused on characterizing ECM components in mature vascular tissue; however, the developing fetal ECM matches the active environment required in vascular tissue engineering more closely. Consequently, we characterized the ECM protein composition of active (fetal) and quiescent (mature) renal arteries using a proteome analysis of decellularized tissue. The obtained human fetal renal artery ECM proteome dataset contains higher levels of 15 ECM proteins versus the mature renal artery ECM proteome, whereas 16 ECM proteins showed higher levels in the mature tissue compared to fetal. Elastic ECM proteins EMILIN1 and FBN1 are significantly enriched in fetal renal arteries and are mainly produced by cells of mesenchymal origin. We functionally tested the role of EMILIN1 and FBN1 by anchoring the ECM secreted by vascular smooth muscle cells (SMCs) to glass coverslips. This ECM layer was depleted from either EMILIN1 or FBN1 by using siRNA targeting of the SMCs. Cultured endothelial cells (ECs) on this modified ECM layer showed alterations on the transcriptome level of multiple pathways, especially the Rho GTPase controlled pathways. However, no significant alterations in adhesion, migration or proliferation were observed when ECs were cultured on EMILIN1- or FNB1-deficient ECM. To conclude, the proteome analysis identified unique ECM proteins involved in the embryonic development of renal arteries. Alterations in transcriptome levels of ECs cultured on EMILIN1- or FBN1-deficient ECM showed that these candidate proteins could affect the endothelial (regenerative) response.
KW - EMILIN1
KW - Extracellular matrix
KW - FBN1
KW - Proteomics
KW - Vasculature
KW - vasculature
KW - proteomics
KW - extracellular matrix
UR - http://www.scopus.com/inward/record.url?scp=85088413420&partnerID=8YFLogxK
U2 - 10.3390/ijms21113905
DO - 10.3390/ijms21113905
M3 - Article
C2 - 32486169
SN - 1422-0067
VL - 21
JO - International journal of molecular sciences
JF - International journal of molecular sciences
IS - 11
M1 - 3905
ER -