title: Endothelial nitric oxide synthase induces heat shock protein HSPA6 (HSP70B') in human arterial smooth muscle cells.
abstract: Endothelial nitric oxide synthase (eNOS) is the major source of nitric oxide (NO) production in blood vessels. One of the pleitropic functions of eNOS derived NO is to inhibit vascular smooth muscle cell proliferation in the blood vessel wall, and whose dysfunction is a primary cause of atherosclerosis and restenosis. In this study there was an interest in examining the gene profile of eNOS adenoviral (Ad-eNOS) transduced human coronary artery smooth muscle cells (HCASMC) to further understand the eNOS inhibitory effect on smooth muscle cell proliferation. To this aim a whole genome wide analysis of eNOS transduced HCASMCs was performed. A total of 19 genes were up regulated, and 31 genes down regulated in Ad-eNOS transduced HCASMCs compared to cells treated with an empty adenovirus. Noticeably, a cluster of HSP70 gene family members was amongst the genes up regulated. Quantitative PCR confirmed that transcripts for HSPA1A (HSP70A), HSPA1B (HSP70B) and HSPA6 (HSP70B') were elevated 2, 1.7 and 14-fold respectively in Ad-eNOS treated cells. The novel gene HSPA6 was further explored as a potential mediator of eNOS signaling in HCASMC. Immunoblotting showed that HSPA6 protein was induced by Ade-NOS. To functionally examine the effect of HSPA6 on SMCs, an adenovirus harboring the HSPA6 gene under the control of a constitutive promoter was generated. Transduction of HCASMCs with Ad-HSPA6 inhibited SMC proliferation at 3 and 6 days post serum growth stimulation, and paralleled the Ad-eNOS inhibition of SMC growth. The identification in this study that HSPA6 overexpression inhibits SMC proliferation coupled with the recent finding that inhibition of HSP90 has a similar effect, progresses the field of targeting HSPs for vascular repair.
authors.affiliation: Regenerative Medicine Institute, National University of Ireland, Galway, Ireland; Department of Physiology, School of Medicine, University Road, National University of Ireland, Galway, Ireland. Electronic address: karl.mccullagh@nuigalway.ie.
authors.forename: K J A
authors.initials: KJ
authors.lastname: McCullagh
authors.affiliation: Regenerative Medicine Institute, National University of Ireland, Galway, Ireland.
authors.forename: R
authors.initials: R
authors.lastname: Cooney
authors.affiliation: Regenerative Medicine Institute, National University of Ireland, Galway, Ireland.
authors.forename: T
authors.initials: T
authors.lastname: O'Brien
mesh_terms: D001158:Arteries / Q000166:cytology*; D002478:Cells, Cultured; D018840:HSP70 Heat-Shock Proteins / Q000096:biosynthesis* / Q000235:genetics; D006801:Humans; D032389:Myocytes, Smooth Muscle / Q000378:metabolism*; D052250:Nitric Oxide Synthase Type III / Q000235:genetics / Q000378:metabolism*
chemical_list: D018840:HSP70 Heat-Shock Proteins; C544598:HSPA6 protein, human; D052250:Nitric Oxide Synthase Type III
keywords: Adenoviral gene transfer; Heat shock protein 70; Human vascular smooth muscle; Microarray; Nitric oxide