TY - JOUR
T1 - Adipose-derived mesenchymal stem cells from patients with atherosclerotic renovascular disease have increased DNA damage and reduced angiogenesis that can be modified by hypoxia
AU - Saad, Ahmed
AU - Zhu, Xiang Yang
AU - Herrmann, Sandra
AU - Hickson, Latonya
AU - Tang, Hui
AU - Dietz, Allan B.
AU - Van Wijnen, Andre J.
AU - Lerman, Lilach
AU - Textor, Stephen
N1 - Funding Information:
This project was partly supported by National Institutes of Health (NIH) grants, including P01 HL85307 from the National Heart, Lung and Blood Institute (NHLBI), R01 DK100081, DK10232, and R01 DK 73608 from the National Institute for Digestive, Diabetic and Kidney Diseases (NIDDK), as well as Clinical and Translational Science Award (CTSA) Grant UL1 RR024150 from NIH/ National Center for Research Resources (NCRR). Our studies were also supported by funds from the Center of Regenerative Medicine at Mayo Clinic. Furthermore, we appreciate the generous philanthropic support of William and Karen Eby, as well as the charitable foundation in their names. The content is solely the responsibility of the authors and does not represent the official views of the NHLBI, NIDDK or the National Institutes of Health.
Publisher Copyright:
© 2016 The Author(s).
PY - 2016/9/9
Y1 - 2016/9/9
N2 - Background: Adipose-derived MSC (AMSCs) possess angiogenic and immunomodulatory properties that may modulate kidney regeneration. Whether these properties are retained in older patients with atherosclerotic vascular disease is poorly understood. Hypoxic conditions are known to modify properties and growth characteristics of AMSCs. We tested the hypothesis that AMSCs from older patients with atherosclerotic renovascular disease (RVD) differ from normal kidney donors, and whether hypoxia changes their functional and molecular properties to promote angiogenesis. Methods: AMSCs from 11 patients with RVD (mean age =74.5 years) and 10 healthy kidney donors (mean age = 51.2 years) were cultured under normoxia (20 % O2) and hypoxia (1 % O2) for 3-4 days until they reached 80 % confluency. We analyzed expression of genes and microRNAs using RNA sequencing and real-time quantitative rt-PCR. Protein expression of selected angiogenic factors (VEGF, IGF, HGF and EGF) were quantified in conditioned media using ELISAs. Apoptosis was tested using Annexin IV staining. Results: Normoxic AMSC from RVD patients grew normally, but exhibited increased DNA damage and reduced migration. VEGF protein secretion was significantly lower in the RVD AMSCs (0.08 vs 2.4 ng/mL/ cell, p <0.05) while HGF was higher. Both trends were reversed during growth under hypoxic conditions. Hypoxia upregulated pro-angiogenic mRNAs expression in AMSCs (VEGF, FGF, STC and ANGPTL4), and downregulated expression of many miRNAs (e.g., miR-15a, miR-16, miR-93, miR-424, 126, 132, 221) except miR-210. Conclusions: Thus, although AMSC from patients with RVD had increased DNA damage and reduced migration, hypoxia stimulated pro-angiogenic responses via increased expression of angiogenic genes, VEGF secretion and induction of the hypoxia-inducible miR-210, while downregulating angiogenesis-related miRNAs.
AB - Background: Adipose-derived MSC (AMSCs) possess angiogenic and immunomodulatory properties that may modulate kidney regeneration. Whether these properties are retained in older patients with atherosclerotic vascular disease is poorly understood. Hypoxic conditions are known to modify properties and growth characteristics of AMSCs. We tested the hypothesis that AMSCs from older patients with atherosclerotic renovascular disease (RVD) differ from normal kidney donors, and whether hypoxia changes their functional and molecular properties to promote angiogenesis. Methods: AMSCs from 11 patients with RVD (mean age =74.5 years) and 10 healthy kidney donors (mean age = 51.2 years) were cultured under normoxia (20 % O2) and hypoxia (1 % O2) for 3-4 days until they reached 80 % confluency. We analyzed expression of genes and microRNAs using RNA sequencing and real-time quantitative rt-PCR. Protein expression of selected angiogenic factors (VEGF, IGF, HGF and EGF) were quantified in conditioned media using ELISAs. Apoptosis was tested using Annexin IV staining. Results: Normoxic AMSC from RVD patients grew normally, but exhibited increased DNA damage and reduced migration. VEGF protein secretion was significantly lower in the RVD AMSCs (0.08 vs 2.4 ng/mL/ cell, p <0.05) while HGF was higher. Both trends were reversed during growth under hypoxic conditions. Hypoxia upregulated pro-angiogenic mRNAs expression in AMSCs (VEGF, FGF, STC and ANGPTL4), and downregulated expression of many miRNAs (e.g., miR-15a, miR-16, miR-93, miR-424, 126, 132, 221) except miR-210. Conclusions: Thus, although AMSC from patients with RVD had increased DNA damage and reduced migration, hypoxia stimulated pro-angiogenic responses via increased expression of angiogenic genes, VEGF secretion and induction of the hypoxia-inducible miR-210, while downregulating angiogenesis-related miRNAs.
KW - Angiogenesis
KW - Hypoxia
KW - Mesenchymal stem cells
KW - Renovascular disease
KW - VEGF and MicroRNAs
UR - http://www.scopus.com/inward/record.url?scp=84986917806&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=84986917806&partnerID=8YFLogxK
U2 - 10.1186/s13287-016-0389-x
DO - 10.1186/s13287-016-0389-x
M3 - Article
C2 - 27612459
AN - SCOPUS:84986917806
SN - 1757-6512
VL - 7
JO - Stem Cell Research and Therapy
JF - Stem Cell Research and Therapy
IS - 1
M1 - 128
ER -