Macro and microvascular diseases are the principal causes of morbidity and mortality in patients with type 1 and type 2 diabetes. Endothelial dysfunction, as evidenced by the increased release of free radicals, increases adhesion molecules and apoptosis. Previous work by us and others has shown that a decrease in HO-1 gene expression enhances apoptosis and vascular injury which can be prevented by pharmacological mediated upregulation of HO-1. Our preliminary results indicate that upregulation of HO-1 prevents endothelial apoptosis by decreasing cellular heme content, increasing CO and bilirubin levels and decreasing EC-SOD and DNA degradation. The goal of this proposal is to elucidate the mechanism by which HO-1 prevents diabetes-mediated endothelial dysfunction and to explore the use of genetic and/or pharmacological approaches to achieve long-term vascular protection. Our hypothesis is that HO-1 gene transfer will provide powerful vascular protection by increasing heme degradation and subsequently increasing CO and bilirubin synthesis. The increased levels of CO and bilirubin will result in an increase in EC-SOD, and eNOS, a decrease in O2- and improved vascular response. In addition, increase in mitochondrial HO-1 levels will increase mitochondrial transport carrier, decrease mitochondrial ROS and prevent release of cytochrome c and activation of caspase 9. We plan to test this hypothesis using genetic probes (retroviral LXSN lentiviral, and adenoviral) and in genetically spontaneously diabetic mice, (NOD) mice. We will use the loss-of-function HO-2 (-/-) and gain-of-function, HO-1 and HO-2 gene transfer to HO-2 (-/-) to decipher the role of each gene in vascular protection. The following specific aims will test the hypothesis. (1)To determine whether genetic intervention, using retroviral vectors, to selectively increases HO-1 provides vascular protection in diabetic rats and NOD mice, and whether this effect is due to CO, bilirubin or both. We will examine the effect HO-1 derived CO and bilirubin on EC-SOD, eNOS and O2- levels. (2) HO-2 (-/-) and NOD mice will be used to test the hypothesis that HO-1 gene transfer can provide vascular protection and that CO, bilirubin or both are obligatory for the vascular protection via increase in EC-SOD, eNOS, decrease in O2- and iNOS. (3)To determine the mechanism whereby HO-1 gene expression (CO and bilirubin) act on the extrinsic and intrinsic pathway of pro-apoptosis and anti-apoptotic proteins. This will examine the role of CO and bilirubin produced by HO-1 in both the mitochondria and cytoplasm. (4) To evaluate whether lentiviral vectors targeting endothelial cells using cell specific promoter (VE-CAD) to overexpress HO-1 gene is sufficient to offset diabetes-induced vascular injury. This proposal is novel in its approach. It will allow for the first time, an in-depth analysis of the function of HO-1 in vascular protection and for the development of innovative gene-targeting therapies for the treatment of type I diabetes.

Agency
National Institute of Health (NIH)
Institute
National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK)
Type
Research Project (R01)
Project #
3R01DK068134-05S1
Application #
8031600
Study Section
Hypertension and Microcirculation Study Section (HM)
Program Officer
Jones, Teresa L Z
Project Start
2010-03-15
Project End
2010-07-31
Budget Start
2010-03-15
Budget End
2010-07-31
Support Year
5
Fiscal Year
2010
Total Cost
$52,645
Indirect Cost
Name
University of Toledo
Department
Physiology
Type
Schools of Medicine
DUNS #
807418939
City
Toledo
State
OH
Country
United States
Zip Code
43614
Puri, Nitin; Zhang, Fan; Monu, Sumit R et al. (2013) Antioxidants condition pleiotropic vascular responses to exogenous H(2)O(2): role of modulation of vascular TP receptors and the heme oxygenase system. Antioxid Redox Signal 18:471-80
Kim, Dong Hyun; Liu, Jiayong; Bhat, Samerna et al. (2013) Peroxisome proliferator-activated receptor delta agonist attenuates nicotine suppression effect on human mesenchymal stem cell-derived osteogenesis and involves increased expression of heme oxygenase-1. J Bone Miner Metab 31:44-52
Burgess, Angela P H; Vanella, Luca; Bellner, Lars et al. (2012) Heme oxygenase (HO-1) rescue of adipocyte dysfunction in HO-2 deficient mice via recruitment of epoxyeicosatrienoic acids (EETs) and adiponectin. Cell Physiol Biochem 29:99-110
Sodhi, Komal; Puri, Nitin; Inoue, Kazuyoshi et al. (2012) EET agonist prevents adiposity and vascular dysfunction in rats fed a high fat diet via a decrease in Bach 1 and an increase in HO-1 levels. Prostaglandins Other Lipid Mediat 98:133-42
Vanella, Luca; Li, Ming; Kim, DongHyun et al. (2012) ApoA1: mimetic peptide reverses adipocyte dysfunction in vivo and in vitro via an increase in heme oxygenase (HO-1) and Wnt10b. Cell Cycle 11:706-14
Burgess, Angela; Vanella, Luca; Bellner, Lars et al. (2012) Epoxyeicosatrienoic acids and heme oxygenase-1 interaction attenuates diabetes and metabolic syndrome complications. Prostaglandins Other Lipid Mediat 97:1-16
Cao, Jian; Peterson, Stephen J; Sodhi, Komal et al. (2012) Heme oxygenase gene targeting to adipocytes attenuates adiposity and vascular dysfunction in mice fed a high-fat diet. Hypertension 60:467-75
Cao, Jian; Inoue, Kazuyoshi; Sodhi, Komal et al. (2012) High-fat diet exacerbates renal dysfunction in SHR: reversal by induction of HO-1-adiponectin axis. Obesity (Silver Spring) 20:945-53
Puri, Nitin; Sodhi, Komal; Haarstad, Michael et al. (2012) Heme induced oxidative stress attenuates sirtuin1 and enhances adipogenesis in mesenchymal stem cells and mouse pre-adipocytes. J Cell Biochem 113:1926-35
Abraham, Nader G; Kappas, Attallah (2011) Mechanism of heme-heme oxygenase system impairment of endothelium contraction in the spontaneously hypertensive rat. Hypertension 58:772-3

Showing the most recent 10 out of 50 publications