title: Increase of ADAM10 level in coronary artery in-stent restenosis segments in diabetic minipigs: high ADAM10 expression promoting growth and migration in human vascular smooth muscle cells via Notch 1 and 3.
abstract: BACKGROUND
  This study aimed to identify major proteins in the pathogenesis of coronary artery in-stent restenosis (ISR) in diabetic minipigs with sirolimus-eluting stenting, and to investigate the roles of key candidate molecules, particularly ADAM10, in human arterial smooth muscle cells (HASMCs).
  
  
  RESULTS
  The stents were implanted in the coronary arteries of 15 diabetic and 26 non-diabetic minipigs, and angiography was repeated at six months. The intima of one vascular segment with significant ISR and one with non-ISR in diabetic minipigs were isolated and cultured in conditioned medium (CM). The CM was analyzed by LC-MS/MS to uncover proteins whose levels were significantly increased (≥ 1.5-fold) in ISR than in non-ISR tissues. After literature searching, we focused on the identified proteins, whose biological functions were most potentially related to ISR pathophysiology. Among them, ADAM10 was significantly increased in diabetic and non-diabetic ISR tissues as compared with non-ISR controls. In cell experiments, retrovirus-mediated overexpression of ADAM10 promoted growth and migration of HASMCs. The effects of ADAM10 were more remarkable in high-glucose culture than in low-glucose culture. Using shRNA and an inhibitor of γ-secretase (GSI), we found that the influences of ADAM10 were in part mediated by Notch1 and notch 3 pathway, which up-regulated Notch downstream genes and enhanced nuclear translocation of the small intracellular component of Notch1 and Notch3.
  
  
  CONCLUSIONS
  This study has identified significantly increased expression of ADAM10 in the ISR versus non-ISR segment in diabetic minipigs and implicates ADAM10 in the enhanced neointimal formation observed in diabetes after vascular injury.
authors.affiliation: Institute of Cardiovascular Diseases, Medical School of Jiaotong University, Shanghai, People's Republic of China.
authors.forename: Ke
authors.initials: K
authors.lastname: Yang
authors.affiliation: Institute of Cardiovascular Diseases, Medical School of Jiaotong University, Shanghai, People's Republic of China ; Department of Cardiology, Rui Jin Hospital, Medical School of Jiaotong University, Shanghai, People's Republic of China.
authors.forename: Lin
authors.initials: L
authors.lastname: Lu
authors.affiliation: Institute of Cardiovascular Diseases, Medical School of Jiaotong University, Shanghai, People's Republic of China.
authors.forename: Yan
authors.initials: Y
authors.lastname: Liu
authors.affiliation: Institute of Cardiovascular Diseases, Medical School of Jiaotong University, Shanghai, People's Republic of China ; Department of Cardiology, Rui Jin Hospital, Medical School of Jiaotong University, Shanghai, People's Republic of China.
authors.forename: Qi
authors.initials: Q
authors.lastname: Zhang
authors.affiliation: Institute of Cardiovascular Diseases, Medical School of Jiaotong University, Shanghai, People's Republic of China.
authors.forename: Li Jin
authors.initials: LJ
authors.lastname: Pu
authors.affiliation: Institute of Cardiovascular Diseases, Medical School of Jiaotong University, Shanghai, People's Republic of China ; Department of Cardiology, Rui Jin Hospital, Medical School of Jiaotong University, Shanghai, People's Republic of China.
authors.forename: Lin Jie
authors.initials: LJ
authors.lastname: Wang
authors.affiliation: Institute of Cardiovascular Diseases, Medical School of Jiaotong University, Shanghai, People's Republic of China ; Department of Cardiology, Rui Jin Hospital, Medical School of Jiaotong University, Shanghai, People's Republic of China.
authors.forename: Zhen Bing
authors.initials: ZB
authors.lastname: Zhu
authors.affiliation: Institute of Cardiovascular Diseases, Medical School of Jiaotong University, Shanghai, People's Republic of China.
authors.forename: Ya Nan
authors.initials: YN
authors.lastname: Wang
authors.affiliation: Institute of Cardiovascular Diseases, Medical School of Jiaotong University, Shanghai, People's Republic of China.
authors.forename: Hua
authors.initials: H
authors.lastname: Meng
authors.affiliation: Institute of Cardiovascular Diseases, Medical School of Jiaotong University, Shanghai, People's Republic of China.
authors.forename: Xiao Jie
authors.initials: XJ
authors.lastname: Zhang
authors.affiliation: Department of Cardiology, Rui Jin Hospital, Medical School of Jiaotong University, Shanghai, People's Republic of China.
authors.forename: Run
authors.initials: R
authors.lastname: Du
authors.affiliation: Institute of Cardiovascular Diseases, Medical School of Jiaotong University, Shanghai, People's Republic of China.
authors.forename: Qiu Jing
authors.initials: QJ
authors.lastname: Chen
authors.affiliation: Institute of Cardiovascular Diseases, Medical School of Jiaotong University, Shanghai, People's Republic of China ; Department of Cardiology, Rui Jin Hospital, Medical School of Jiaotong University, Shanghai, People's Republic of China.
authors.forename: Wei Feng
authors.initials: WF
authors.lastname: Shen
mesh_terms: D051722:ADAM Proteins / Q000235:genetics / Q000378:metabolism*; D000072197:ADAM10 Protein; D053829:Amyloid Precursor Protein Secretases / Q000235:genetics / Q000378:metabolism*; D000818:Animals; D002465:Cell Movement; D049109:Cell Proliferation; D023903:Coronary Restenosis / Q000378:metabolism* / Q000473:pathology; D003331:Coronary Vessels / Q000378:metabolism* / Q000473:pathology; D048909:Diabetes Complications / Q000378:metabolism / Q000473:pathology; D015870:Gene Expression; D006801:Humans; D006965:Hyperplasia; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism*; D009131:Muscle, Smooth, Vascular / Q000166:cytology*; D058426:Neointima / Q000473:pathology; D040901:Proteomics; D051881:Receptor, Notch1 / Q000378:metabolism*; D015398:Signal Transduction; D015607:Stents; D013552:Swine; D013556:Swine, Miniature
chemical_list: D008565:Membrane Proteins; D051881:Receptor, Notch1; D053829:Amyloid Precursor Protein Secretases; D051722:ADAM Proteins; D000072197:ADAM10 Protein; C491236:ADAM10 protein, human