title: Increased endogenous H2S generation by CBS, CSE, and 3MST gene therapy improves ex vivo renovascular relaxation in hyperhomocysteinemia.
abstract: Hydrogen sulfide (H(2)S) has recently been identified as a regulator of various physiological events, including vasodilation, angiogenesis, antiapoptotic, and cellular signaling. Endogenously, H(2)S is produced as a metabolite of homocysteine (Hcy) by cystathionine β-synthase (CBS), cystathionine γ-lyase (CSE), and 3-mercaptopyruvate sulfurtransferase (3MST). Although Hcy is recognized as vascular risk factor at an elevated level [hyperhomocysteinemia (HHcy)] and contributes to vascular injury leading to renovascular dysfunction, the exact mechanism is unclear. The goal of the current study was to investigate whether conversion of Hcy to H(2)S improves renovascular function. Ex vivo renal artery culture with CBS, CSE, and 3MST triple gene therapy generated more H(2)S in the presence of Hcy, and these arteries were more responsive to endothelial-dependent vasodilation compared with nontransfected arteries treated with high Hcy. Cross section of triple gene-delivered renal arteries immunostaining suggested increased expression of CD31 and VEGF and diminished expression of the antiangiogenic factor endostatin. In vitro endothelial cell culture demonstrated increased mitophagy during high levels of Hcy and was mitigated by triple gene delivery. Also, dephosphorylated Akt and phosphorylated FoxO3 in HHcy were reversed by H(2)S or triple gene delivery. Upregulated matrix metalloproteinases-13 and downregulated tissue inhibitor of metalloproteinase-1 in HHcy were normalized by overexpression of triple genes. Together, these results suggest that H(2)S plays a key role in renovasculopathy during HHcy and is mediated through Akt/FoxO3 pathways. We conclude that conversion of Hcy to H(2)S by CBS, CSE, or 3MST triple gene therapy improves renovascular function in HHcy.
authors.affiliation: Department of Physiology and Biophysics, University of Louisville School of Medicine, Kentucky, USA. u0sen001@louisville.edu
authors.forename: Utpal
authors.initials: U
authors.lastname: Sen
authors.forename: Pushpakumar B
authors.initials: PB
authors.lastname: Sathnur
authors.forename: Sourav
authors.initials: S
authors.lastname: Kundu
authors.forename: Srikanth
authors.initials: S
authors.lastname: Givvimani
authors.forename: Denise M
authors.initials: DM
authors.lastname: Coley
authors.forename: Paras K
authors.initials: PK
authors.lastname: Mishra
authors.forename: Natia
authors.initials: N
authors.lastname: Qipshidze
authors.forename: Neetu
authors.initials: N
authors.lastname: Tyagi
authors.forename: Naira
authors.initials: N
authors.lastname: Metreveli
authors.forename: Suresh C
authors.initials: SC
authors.lastname: Tyagi
mesh_terms: D000818:Animals; D002478:Cells, Cultured; D003541:Cystathionine beta-Synthase / Q000235:genetics* / Q000378:metabolism; D003542:Cystathionine gamma-Lyase / Q000235:genetics* / Q000378:metabolism; D043169:Endostatins / Q000096:biosynthesis; D000071316:Forkhead Box Protein O3; D051858:Forkhead Transcription Factors / Q000378:metabolism; D015316:Genetic Therapy; D006710:Homocysteine / Q000378:metabolism; D006862:Hydrogen Sulfide / Q000378:metabolism*; D020138:Hyperhomocysteinemia / Q000235:genetics / Q000378:metabolism / Q000628:therapy*; D006978:Hypertension, Renovascular / Q000235:genetics / Q000628:therapy; D053509:Matrix Metalloproteinase 13 / Q000378:metabolism; D051379:Mice; D008810:Mice, Inbred C57BL; D009924:Organ Culture Techniques; D019408:Platelet Endothelial Cell Adhesion Molecule-1 / Q000096:biosynthesis; D051057:Proto-Oncogene Proteins c-akt / Q000378:metabolism; D017382:Reactive Oxygen Species / Q000378:metabolism; D012077:Renal Artery / Q000378:metabolism; D013466:Sulfurtransferases / Q000235:genetics* / Q000378:metabolism; D019715:Tissue Inhibitor of Metalloproteinase-1 / Q000378:metabolism; D042461:Vascular Endothelial Growth Factor A / Q000096:biosynthesis; D057772:Vascular System Injuries
chemical_list: D043169:Endostatins; D000071316:Forkhead Box Protein O3; D051858:Forkhead Transcription Factors; C474033:FoxO3 protein, mouse; D019408:Platelet Endothelial Cell Adhesion Molecule-1; D017382:Reactive Oxygen Species; D019715:Tissue Inhibitor of Metalloproteinase-1; D042461:Vascular Endothelial Growth Factor A; D006710:Homocysteine; D051057:Proto-Oncogene Proteins c-akt; D013466:Sulfurtransferases; C021550:3-mercaptopyruvate sulphurtransferase; D053509:Matrix Metalloproteinase 13; D003541:Cystathionine beta-Synthase; D003542:Cystathionine gamma-Lyase; D006862:Hydrogen Sulfide