Formation of Hyaluronan- and Versican-Rich Pericellular Matrix Is Required for Proliferation and Migration of Vascular Smooth Muscle Cells

Abstract

The accumulation of hyaluronan (HA) and the HA-binding proteoglycan versican around smooth muscle cells in lesions of atherosclerosis suggests that together these molecules play an important role in the events of atherogenesis. In this study we have examined the formation of HA- and versican-rich pericellular matrices by human aortic smooth muscle cells in vitro, using a particle-exclusion assay, and the role of the pericellular matrix in cell proliferation and migration. The structural dependence of the pericellular matrix on HA can be demonstrated by the complete removal of the matrix with Streptomyces hyaluronidase. The presence of versican in the pericellular matrix was confirmed immunocytochemically. By electron microscopy, the cell coat was seen as a tangled network of hyaluronidase-sensitive filaments decorated with ruthenium red-positive proteoglycan granules. Ninety percent of migrating cells in wounded cultures, and virtually all mitotic cells, displayed abundant HA- and versican-rich coats. Time-lapse video imaging revealed that HA- and versican-rich pericellular matrix formation is dynamic and rapid, and coordinated specifically with cell detachment and mitotic cell rounding. HA oligosaccharides, which inhibit the binding of HA to the cell surface and prevent pericellular matrix formation, significantly reduced proliferation and migration in response to platelet-derived growth factor, whereas larger HA fragments and high molecular weight HA had no effect. Treatment with HA oligosaccharides also led to changes in cell shape from a typical fusiform morphology to a more spread and flattened appearance. These data suggest that organization of HA- and versican-rich pericellular matrices may facilitate migration and mitosis by diminishing cell surface adhesivity and affecting cell shape through steric exclusion and the viscous properties of HA proteoglycan gels.

Versican Degradation and Vascular Disease

“…The converse experiments using antisense or siRNA also indicate that endogenous versican V1 increases proliferation, including that of SMCs , Huang et al 2006) (unpublished data, Rahmani M, Wong B, Allahverdian S, Cheung C, Carthy J, Keire P, Wight T, McManus B). These data are consistent with evidence that formation of versican-hyaluronan pericellular matrix is required for SMC proliferation and migration (Evanko et al 1999). Versican V1 may enhance proliferation via increased levels of the EGF receptor and ERK activation coupled with loss of the cell-cycle inhibitor p27 (unpublished data, Rahmani M, Wong B, Allahverdian S, Cheung C, Carthy J, Keire P, Wight T, McManus B) (Sheng et al 2005).…”

Antisense inhibition of hyaluronan synthase-2 in human osteosarcoma cells inhibits hyaluronan retention and tumorigenicity

“…Assembly of HA-rich, hydrated matrices around the cells stimulates migration [32] and tumor cell-HA interactions mediated through CD44 may play roles in facilitating migration through the tumor associated HA-rich matrix [3]. Formation of hydrated, HA-rich matrix was disrupted in antisense treated cells, as visualized by the particle exclusion assay, resulting in reduced motility and invasiveness in this study.…”

Proliferative Effect of Lipoprotein Lipase on Human Vascular Smooth Muscle Cells

“…[41][42][43] Our finding that treatment of VSMCs with heparinase suppresses LPL-induced VSMC proliferation suggests that LPL binding to HSPGs expressed on the VSMC surface is essential for its effect on cell proliferation. However, because the formation of a proteoglycan-rich pericellular matrix may be involved in VSMC proliferation, 44 inhibition of VSMC growth by heparinase treatment of the cells could also be due to alteration of the extracellular matrix integrity. Recently, Silver et al 45 showed that locally delivered heparinase reduced medial VSMC proliferation induced by balloon catheter injury in rat carotid arteries.…”