Caspases from apoptotic myocytes degrade extracellular matrix: a novel remodeling paradigm.

Induction of smooth muscle cell apoptosis is critical to the reversal of severe structural remodeling in hypertensive pulmonary arteries during disease regression. This process involves coordinated resorption of pathologically deposited extracellular matrix, including elastin, and occurs in the presence of serine elastase and matrix metalloproteinase inhibitors. Here, we show that apoptotic smooth muscle cells exhibit extensive degradation of elastin coincident with cell surface immunolocalization and release of caspases. We further document that recombinant caspase-2, -3, and -7 are potently elastolytic. These enzymes are present in an active form on apoptotic cell surfaces and caspase inhibitors attenuate their elastolytic activity. Our results reveal a previously undescribed function for apoptotic cells and a novel paradigm whereby removal of cells is coordinated with degradation of excess extracellular matrix during remodeling in development and disease.

Apolipoprotein D and platelet-derived growth factor-BB synergism mediates vascular smooth muscle cell migration.

We identified apolipoprotein (apo)D in a search for proteins upregulated in a posttranscriptional manner similar to fibronectin in motile smooth muscle cells (SMCs). To address the function of apoD in SMCs, we cloned a partial apoD cDNA from ovine aortic (Ao) SMCs using RT-PCR. We documented a 2.5-fold increase in apoD protein but no increase in apoD mRNA in Ao SMCs 48 hours after a multiwound migration assay (P<0.01). Confocal microscopy revealed prominent perinuclear and trailing edge expression of apoD in migrating SMCs but not in the confluent monolayer. Stimulation of Ao SMCs with 10 ng/mL platelet-derived growth factor (PDGF)-BB increased apoD protein expression (P<0.05). Moreover, PDGF-BB-stimulated migration of human pulmonary artery SMCs was suppressed by knock-down of apoD using RNAi. Stable overexpression of apoD in Ao SMCs cultured in 10% fetal bovine serum promoted random migration by 62% compared with vector-transfected cells (P<0.01). Overexpression of apoD or addition of exogenous apoD to a rat aortic SMC line (A10) stimulated their migration in response to a subthreshold dose of PDGF-BB (P<0.05). This was unrelated to increased phosphorylation of ERK1/2 or of phospholipase C-gamma1, but correlated with enhanced Rac1 activation. This study shows that apoD can be expressed or taken up by SMCs and can regulate their motility in response to growth factors.