Lentivirus production

Atherosclerosis is the most common underlying cause of cardiovascular morbidity and mortality worldwide. c-Kit (CD117) is a member of the receptor tyrosine kinase family, which regulates differentiation, proliferation, and survival of multiple cell types. Recent studies have shown that c-Kit and its ligand stem cell factor (SCF) are present in arterial endothelial cells and smooth muscle cells (SMCs). The role of c-Kit in cardiovascular disease remains unclear. The aim of the current study is to determine the role of c-Kit in atherogenesis. For this purpose, atherosclerotic plaques were quantified in c-Kit deficient mice (Kit^Mut) after feeding them a high fat diet (HFD) for 16 weeks. Kit^Mutmicedemonstrated substantially greater atherosclerosis compared with control (Kit^WT) littermates (p<0.01). Transplantation of c-Kit positive bone marrow into Kit^Mutmice failed to rescue the atherogenic phenotype: an indication that increased atherosclerosis was associated with reduced arterial c-Kit. To investigate the mechanism, SMC organization and morphology were analyzed in the aorta by histopathology and electron microscopy. SMCs were more abundant, disorganized, and vacuolated in aortas of c-Kit mutant mice compared with controls (p<0.05). Markers of the "contractile" SMC phenotype (calponin, SM22α) were downregulated with pharmacologic and genetic c-Kit inhibition (p<0.05). The absence of c-Kit increased lipid accumulation, and significantly reduced the expression of the ATP-binding cassette transporter G1 (ABCG1) necessary for lipid efflux in SMCs. Reconstitution of c-Kit in cultured Kit^MutSMCs resulted in increased spindle-shaped morphology, reduced proliferation, and elevated levels of contractile markers; all indicators of their restored contractile phenotype (p<0.05).