P66ShcA modulates oxidative stress and survival of endothelial progenitor cells in response to high glucose

Valeria Di Stefano, Chiara Cencioni, Germana Zaccagnini, Alessandra Magenta, Maurizio C. Capogrossi, Fabio Martelli

Research output: Contribution to journalArticle

49 Citations (Scopus)

Abstract

AimsA close relationship exists between hyperglycaemia, oxidative stress, and diabetic complications. In fact, high glucose (HG) determines the overproduction of reactive oxygen species (ROS) by the mitochondria. p66 ShcA is a gene that regulates the apoptotic responses to oxidative stress. Indeed, p66ShcA knockout (ko) mice display decreased ROS production and increased resistance to ROS-induced cell death in a variety of pathophysiological settings. Reduced endothelial progenitor cell (EPC) number, differentiation, and function are relevant components of the angiogenesis impairment observed in diabetic patients. We examined the role of p66 ShcA in the EPC deficit induced by HG.Methods and resultsMouse bone marrow-derived c-kit+ cells differentiate in endothelial-like cells when plated on fibronectin (BM-derived EPCs). We found that cell culture in the presence of HG up-regulated p66ShcA protein expression and that HG exposure markedly decreased the number of BM-derived EPCs. Conversely, p66 ShcA ko BM-derived EPCs were not sensitive to HG inhibition. Indeed, the resistance of p66ShcA ko BM-derived EPCs to HG was associated with reduced levels of both apoptosis and oxidative stress. To functionally link the HG response to ROS production, p66ShcA ko BM-derived EPCs were reconstituted either with p66ShcA wild-type (wt) or with a p66 ShcA allele (p66ShcA qq) that was devoid of its ROS-generating function. We found that only p66ShcA wt and not the qq mutant rescued p66ShcA ko cell sensitivity to HG. One major feature of oxidative stress is its ability to reduce the bio-availability of nitric oxide (NO) that, in turn, plays a crucial role in endothelial differentiation and function. We found that the p66ShcA deletion prevented the HG-induced increase of nitrotyrosine, and that the resistance to HG of p66 ShcA ko BM-derived EPCs was prevented by NO synthase inhibition. With a reciprocal approach, the treatment of p66ShcA wt cells with a NO donor prevented the HG-induced deficit. Finally, using a Matrigel plug angiogenesis assay, we demonstrated that p66ShcA ko prevented diabetic impairment of angiogenesis in vivo.Conclusionp66ShcA deletion rescues the BM-derived EPCs defect induced by HG, indicating p66 ShcA as a potential therapeutic target in diabetic vasculopathy.

Original languageEnglish
Pages (from-to)421-429
Number of pages9
JournalCardiovascular Research
Volume82
Issue number3
DOIs
Publication statusPublished - Jun 2009

Fingerprint

Oxidative Stress
Glucose
Reactive Oxygen Species
Endothelial Progenitor Cells
Nitric Oxide Donors
Diabetes Complications
erucylphosphocholine
Fibronectins
Knockout Mice
Nitric Oxide Synthase
Hyperglycemia
Cell Differentiation
Mitochondria
Nitric Oxide
Cell Death
Endothelial Cells
Cell Culture Techniques
Cell Count
Bone Marrow
Alleles

Keywords

  • Apoptosis
  • Diabetes
  • Endothelium
  • Oxygen radicals
  • Stem cells

ASJC Scopus subject areas

  • Cardiology and Cardiovascular Medicine
  • Physiology (medical)
  • Physiology

Cite this

P66ShcA modulates oxidative stress and survival of endothelial progenitor cells in response to high glucose. / Di Stefano, Valeria; Cencioni, Chiara; Zaccagnini, Germana; Magenta, Alessandra; Capogrossi, Maurizio C.; Martelli, Fabio.

In: Cardiovascular Research, Vol. 82, No. 3, 06.2009, p. 421-429.

Research output: Contribution to journalArticle

Di Stefano, Valeria ; Cencioni, Chiara ; Zaccagnini, Germana ; Magenta, Alessandra ; Capogrossi, Maurizio C. ; Martelli, Fabio. / P66ShcA modulates oxidative stress and survival of endothelial progenitor cells in response to high glucose. In: Cardiovascular Research. 2009 ; Vol. 82, No. 3. pp. 421-429.
@article{8c2a444cbc214500917deb9a8a5527f0,
title = "P66ShcA modulates oxidative stress and survival of endothelial progenitor cells in response to high glucose",
abstract = "AimsA close relationship exists between hyperglycaemia, oxidative stress, and diabetic complications. In fact, high glucose (HG) determines the overproduction of reactive oxygen species (ROS) by the mitochondria. p66 ShcA is a gene that regulates the apoptotic responses to oxidative stress. Indeed, p66ShcA knockout (ko) mice display decreased ROS production and increased resistance to ROS-induced cell death in a variety of pathophysiological settings. Reduced endothelial progenitor cell (EPC) number, differentiation, and function are relevant components of the angiogenesis impairment observed in diabetic patients. We examined the role of p66 ShcA in the EPC deficit induced by HG.Methods and resultsMouse bone marrow-derived c-kit+ cells differentiate in endothelial-like cells when plated on fibronectin (BM-derived EPCs). We found that cell culture in the presence of HG up-regulated p66ShcA protein expression and that HG exposure markedly decreased the number of BM-derived EPCs. Conversely, p66 ShcA ko BM-derived EPCs were not sensitive to HG inhibition. Indeed, the resistance of p66ShcA ko BM-derived EPCs to HG was associated with reduced levels of both apoptosis and oxidative stress. To functionally link the HG response to ROS production, p66ShcA ko BM-derived EPCs were reconstituted either with p66ShcA wild-type (wt) or with a p66 ShcA allele (p66ShcA qq) that was devoid of its ROS-generating function. We found that only p66ShcA wt and not the qq mutant rescued p66ShcA ko cell sensitivity to HG. One major feature of oxidative stress is its ability to reduce the bio-availability of nitric oxide (NO) that, in turn, plays a crucial role in endothelial differentiation and function. We found that the p66ShcA deletion prevented the HG-induced increase of nitrotyrosine, and that the resistance to HG of p66 ShcA ko BM-derived EPCs was prevented by NO synthase inhibition. With a reciprocal approach, the treatment of p66ShcA wt cells with a NO donor prevented the HG-induced deficit. Finally, using a Matrigel plug angiogenesis assay, we demonstrated that p66ShcA ko prevented diabetic impairment of angiogenesis in vivo.Conclusionp66ShcA deletion rescues the BM-derived EPCs defect induced by HG, indicating p66 ShcA as a potential therapeutic target in diabetic vasculopathy.",
keywords = "Apoptosis, Diabetes, Endothelium, Oxygen radicals, Stem cells",
author = "{Di Stefano}, Valeria and Chiara Cencioni and Germana Zaccagnini and Alessandra Magenta and Capogrossi, {Maurizio C.} and Fabio Martelli",
year = "2009",
month = "6",
doi = "10.1093/cvr/cvp082",
language = "English",
volume = "82",
pages = "421--429",
journal = "Cardiovascular Research",
issn = "0008-6363",
publisher = "Oxford University Press",
number = "3",

}

TY - JOUR

T1 - P66ShcA modulates oxidative stress and survival of endothelial progenitor cells in response to high glucose

AU - Di Stefano, Valeria

AU - Cencioni, Chiara

AU - Zaccagnini, Germana

AU - Magenta, Alessandra

AU - Capogrossi, Maurizio C.

AU - Martelli, Fabio

PY - 2009/6

Y1 - 2009/6

N2 - AimsA close relationship exists between hyperglycaemia, oxidative stress, and diabetic complications. In fact, high glucose (HG) determines the overproduction of reactive oxygen species (ROS) by the mitochondria. p66 ShcA is a gene that regulates the apoptotic responses to oxidative stress. Indeed, p66ShcA knockout (ko) mice display decreased ROS production and increased resistance to ROS-induced cell death in a variety of pathophysiological settings. Reduced endothelial progenitor cell (EPC) number, differentiation, and function are relevant components of the angiogenesis impairment observed in diabetic patients. We examined the role of p66 ShcA in the EPC deficit induced by HG.Methods and resultsMouse bone marrow-derived c-kit+ cells differentiate in endothelial-like cells when plated on fibronectin (BM-derived EPCs). We found that cell culture in the presence of HG up-regulated p66ShcA protein expression and that HG exposure markedly decreased the number of BM-derived EPCs. Conversely, p66 ShcA ko BM-derived EPCs were not sensitive to HG inhibition. Indeed, the resistance of p66ShcA ko BM-derived EPCs to HG was associated with reduced levels of both apoptosis and oxidative stress. To functionally link the HG response to ROS production, p66ShcA ko BM-derived EPCs were reconstituted either with p66ShcA wild-type (wt) or with a p66 ShcA allele (p66ShcA qq) that was devoid of its ROS-generating function. We found that only p66ShcA wt and not the qq mutant rescued p66ShcA ko cell sensitivity to HG. One major feature of oxidative stress is its ability to reduce the bio-availability of nitric oxide (NO) that, in turn, plays a crucial role in endothelial differentiation and function. We found that the p66ShcA deletion prevented the HG-induced increase of nitrotyrosine, and that the resistance to HG of p66 ShcA ko BM-derived EPCs was prevented by NO synthase inhibition. With a reciprocal approach, the treatment of p66ShcA wt cells with a NO donor prevented the HG-induced deficit. Finally, using a Matrigel plug angiogenesis assay, we demonstrated that p66ShcA ko prevented diabetic impairment of angiogenesis in vivo.Conclusionp66ShcA deletion rescues the BM-derived EPCs defect induced by HG, indicating p66 ShcA as a potential therapeutic target in diabetic vasculopathy.

AB - AimsA close relationship exists between hyperglycaemia, oxidative stress, and diabetic complications. In fact, high glucose (HG) determines the overproduction of reactive oxygen species (ROS) by the mitochondria. p66 ShcA is a gene that regulates the apoptotic responses to oxidative stress. Indeed, p66ShcA knockout (ko) mice display decreased ROS production and increased resistance to ROS-induced cell death in a variety of pathophysiological settings. Reduced endothelial progenitor cell (EPC) number, differentiation, and function are relevant components of the angiogenesis impairment observed in diabetic patients. We examined the role of p66 ShcA in the EPC deficit induced by HG.Methods and resultsMouse bone marrow-derived c-kit+ cells differentiate in endothelial-like cells when plated on fibronectin (BM-derived EPCs). We found that cell culture in the presence of HG up-regulated p66ShcA protein expression and that HG exposure markedly decreased the number of BM-derived EPCs. Conversely, p66 ShcA ko BM-derived EPCs were not sensitive to HG inhibition. Indeed, the resistance of p66ShcA ko BM-derived EPCs to HG was associated with reduced levels of both apoptosis and oxidative stress. To functionally link the HG response to ROS production, p66ShcA ko BM-derived EPCs were reconstituted either with p66ShcA wild-type (wt) or with a p66 ShcA allele (p66ShcA qq) that was devoid of its ROS-generating function. We found that only p66ShcA wt and not the qq mutant rescued p66ShcA ko cell sensitivity to HG. One major feature of oxidative stress is its ability to reduce the bio-availability of nitric oxide (NO) that, in turn, plays a crucial role in endothelial differentiation and function. We found that the p66ShcA deletion prevented the HG-induced increase of nitrotyrosine, and that the resistance to HG of p66 ShcA ko BM-derived EPCs was prevented by NO synthase inhibition. With a reciprocal approach, the treatment of p66ShcA wt cells with a NO donor prevented the HG-induced deficit. Finally, using a Matrigel plug angiogenesis assay, we demonstrated that p66ShcA ko prevented diabetic impairment of angiogenesis in vivo.Conclusionp66ShcA deletion rescues the BM-derived EPCs defect induced by HG, indicating p66 ShcA as a potential therapeutic target in diabetic vasculopathy.

KW - Apoptosis

KW - Diabetes

KW - Endothelium

KW - Oxygen radicals

KW - Stem cells

UR - http://www.scopus.com/inward/record.url?scp=66249112517&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=66249112517&partnerID=8YFLogxK

U2 - 10.1093/cvr/cvp082

DO - 10.1093/cvr/cvp082

M3 - Article

VL - 82

SP - 421

EP - 429

JO - Cardiovascular Research

JF - Cardiovascular Research

SN - 0008-6363

IS - 3

ER -