Characterization of primary and restenotic atherosclerotic plaque from the superficial femoral artery: Potential role of Smad3 in regulation of SMC proliferation

ObjectiveTo characterize and compare primary and restenotic lesions of the superficial femoral artery and analyze the contribution of TGF-β/Smad3 signaling to the pathophysiology of peripheral artery occlusive disease.Methods and ResultsImmunohistochemical studies were performed on specimens retrieved from the superficial femoral artery of patients undergoing either atherectomy for primary atherosclerotic or recurrent disease after stenting and/or prior angioplasty. Immunohistochemical analysis revealed a significantly higher smooth muscle cell (SMC) content (α-actin+) and expression of Smad3 in restenotic lesions while primary lesions contained significantly more leukocytes (CD45+) and macrophages (CD68+). Further studies demonstrated colocalization of Smad3 with α-actin and PCNA, suggesting a role for Smad3 in the proliferation observed in restenotic lesions. To confirm a role for Smad3 in SMC proliferation, we both upregulated Smad3 via adenoviral mediated gene transfer (AdSmad3) and inhibited Smad3 through transfection with siRNA in human aortic SMCs, then assessed cell proliferation with tritiated thymidine. Overexpression of Smad3 enhanced whereas inhibition of Smad3 decreased cell proliferation.ConclusionDifferences in cellular composition and cell proliferation in conjunction with the finding that Smad3 is expressed exclusively in restenotic disease suggest that TGF-β, through Smad3 signaling, may play an essential role in SMC proliferation and the pathophysiology of restenosis in humans.

Clinical RelevanceIntimal hyperplasia leading to restenosis significantly limits the long-term success of percutaneous interventions for lower extremity vascular occlusive disease. Because the mechanisms leading to the development of restenosis are likely different from those that produce primary atherosclerosis, a comparison of the pathophysiology of restenotic vs primary atherosclerotic plaque will enable us to better define the cellular events that lead to recurrent disease. This will enable us to develop targeted therapies for the prevention of restenosis following vascular intervention for the treatment of peripheral arterial disease.