Circulation Research. 2001
Published online before print November 1, 2001, doi: 10.1161/hh2401.101070
Published online before print November 1, 2001, doi: 10.1161/hh2401.101070
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Submitted on March 20, 2000
Revised on October 19, 2001
Accepted on October 19, 2001
Smooth Muscle Cell Phenotypic Transition Associated With Calcification. Upregulation of Cbfa1 and Downregulation of Smooth Muscle Lineage Markers
From the Departments of Bioengineering (S.A.S., M.Y.S., H.-Y.Y., C.M.G.), Pathology (G.C.), and Molecular Biotechnology (P.H., R.A.), University of Washington, Seattle, Wash; and the Department of Molecular and Human Genetics (T.S., G.K.), Baylor College of Medicine, Houston, Tex. Present address for P.H. is Novartis Agricultural Discovery Institute, San Diego, Calif.
* To whom correspondence should be addressed. E-mail: ceci{at}u.washington.edu.
Bovine aortic smooth muscle cell (BASMC) cultures undergo mineralization on addition of the organic phosphate donor, ß-glycerophosphate (ßGP). Mineralization is characterized by apatite deposition on collagen fibrils and the presence of matrix vesicles, as has been described in calcified vascular lesions in vivo as well as in bone and teeth. In the present study, we used this model to investigate the molecular mechanisms driving vascular calcification. We found that BASMCs lost their lineage markers, SM22
and smooth muscle -actin, within 10 days of being placed under calcifying conditions. Conversely, the cells gained an osteogenic phenotype as indicated by an increase in expression and DNA-binding activity of the transcription factor, core binding factor 1 (Cbfa1). Moreover, genes containing the Cbfa1 binding site, OSE2, including osteopontin, osteocalcin, and alkaline phosphatase were elevated. The relevance of these in vitro findings to vascular calcification in vivo was further studied in matrix GLA protein null (MGP-/-) mice whose arteries spontaneously calcify. We found that arterial calcification was associated with a similar loss in smooth muscle markers and a gain of osteopontin and Cbfa1 expression. These data demonstrate a novel association of vascular calcification with smooth muscle cell phenotypic transition, in which several osteogenic proteins including osteopontin, osteocalcin, and the bone determining factor Cbfa1 are gained. The findings suggest a positive role for SMCs in promoting vascular calcification.
Key words: vascular calcification • smooth muscle cells • phenotype • core binding factor
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