A perlecan-inducing compound significantly inhibits smooth muscle cell function and in-stent intimal hyperplasia: novel insights into the diverse biological effects of perlecan.
Perlecan is the major heparan sulfate proteoglycan in the arterial wall. Previous studies have suggested that perlecan is a potent inhibitor of smooth muscle cell (SMC) activity. Therefore, perlecan overexpression may serve as a therapeutic modality to prevent in-stent restenosis (ISR). We have investigated a novel compound (RUS3108), identified in a SMC based screen to induce perlecan synthesis in SMC. The aims of this study were to assess the in vitro effects of RUS3108 and the effects of RUS3108-eluting stents in preventing ISR.



Rabbit aortic SMC and bovine aortic endothelial cells (EC) were used in this study. Immunohistochemistry showed that RUS3108-treated SMC over-expressed perlecan indicating the drug effects. Furthermore, RUS3108 induced a SMC differentiated phenotype by SMembryonic staining. RUS3108 (1 microM) inhibited 3H-thymidine incorporation by >50%, which was completely reversed by a perlecan antibody. RUS3108 also inhibited SMC migration (Boyden chamber) and MMP-9 activity. In contrast, RUS3108 (100nM) modestly stimulated EC 3H-thymidine incorporation by 22% (p<0.02). In vivo, a total of 30 stents were deployed in rabbit iliac arteries as follows: 1) bare metal stents (n=10), 2) polymer onlycoated stents (n=10), and 3) polymer-coated stents containing RUS3108 (n=10). Rabbits were sacrificed at four weeks and stented segments were subjected to morphometric analysis. Intimal cross sectional area was significantly lower in the RUS3108-eluting stent group (0.31+ or -0.27 mm(2) versus 1.0 + or - 0.31 and 1.25 + or - 0.51 in the bare metal stents and polymer only coated stents groups, respectively, p<0.0001).



RUS3108 is a novel perlecan-inducing compound, which is a potent inhibitor of SMC activity and a modest stimulator of EC proliferation. RUS3108-eluting stents may serve as an excellent modality for the prevention of ISR.
Segev
Amit
A
The Roy and Ann Foss Cardiovascular Research Program, Terrence Donnelly Heart Center, St. Michael's Hospital, University of Toronto, Toronto, Ontario, Canada.
Nili
Nafiseh
N
Osherov
Azriel B
AB
Qiang
Beiping
B
Wong
Amy J
AJ
Pillarisetti
Sivaram
S
Strauss
Bradley H
BH
D000800:Angioplasty, Balloon / Q000009:adverse effects / Q000295:instrumentation*; D000818:Animals; D002317:Cardiovascular Agents / Q000008:administration & dosage*; D002417:Cattle; D002454:Cell Differentiation / Q000187:drug effects; D002465:Cell Movement / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects*; D002478:Cells, Cultured; D004305:Dose-Response Relationship, Drug; D054855:Drug-Eluting Stents; D042783:Endothelial Cells / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology; D019812:Heparan Sulfate Proteoglycans / Q000378:metabolism*; D006965:Hyperplasia; D007083:Iliac Artery / Q000187:drug effects / Q000378:metabolism / Q000473:pathology; D020778:Matrix Metalloproteinase 2 / Q000378:metabolism; D020780:Matrix Metalloproteinase 9 / Q000378:metabolism; D009131:Muscle, Smooth, Vascular / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology; D032389:Myocytes, Smooth Muscle / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology; D011817:Rabbits; D013997:Time Factors; D017539:Tunica Intima / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology; D015854:Up-Regulation
D002317:Cardiovascular Agents; D019812:Heparan Sulfate Proteoglycans; C071927:perlecan; D020778:Matrix Metalloproteinase 2; D020780:Matrix Metalloproteinase 9