This project addresses neurovascular injury during ischemic retinopathy. While this condition is associated with early neurovascular dysfunction, conventional therapies target clinically significant macula edema or neovascularization, which occur much later. Therapy to prevent/reverse ischemic retinal injury is a critical unmet need. The project goal is to delineate mechanisms of vascular and neuronal injury during retinopathy and identify novel therapeutic strategies. The investigators'studies in models of ischemic retinopathy have revealed that the urea cycle enzyme arginase is critically involved in both vascular and neuronal injury. Arginase metabolizes L-arginine to form proline, polyamines and glutamate. Excessive arginase activity reduces the L-arginine supply for nitric oxide synthase (NOS), causing it to become uncoupled and produce superoxide and less NO. Superoxide and NO react rapidly and form the toxic oxidant peroxynitrite. Glutamate and the catabolic products of polyamine oxidation can induce more oxidative stress and DNA damage, both of which can cause mitochondrial injury and premature senescence. Preliminary data show that neurovascular injury during retinopathy is associated with increased arginase expression/activity, decreased NO, polyamine oxidation, increased formation of superoxide and peroxynitrite, mitochondrial injury and premature senescence. Furthermore, the cytosolic isoform arginase 1 (A1) is implicated in premature senescence and dysfunction of vascular endothelial cells (EC), whereas the mitochondrial isoform arginase 2 (A2) appears to be involved in neuronal dysfunction/injury. Thus, it is hypothesized that activation of the arginase pathway causes neurovascular injury by uncoupling NOS and inducing polyamine oxidation and glutamate formation, thereby reducing NO and increasing oxidative stress, leading to mitochondrial dysfunction, EC senescence and vascular and neuronal dysfunction.
Aim 1 will use animal and tissue culture models to test whether (A) limiting A1 expression will prevent vascular dysfunction by blocking NOS uncoupling, reducing oxidative stress and preventing mitochondrial dysfunction and senescence of ECs;(B) limiting A2 expression will prevent neuronal injury by blocking polyamine oxidation and glutamate formation, reducing oxidative stress and preventing mitochondrial and neuronal dysfunction.
Aim 2 will determine the effects on neurovascular dysfunction and injury of novel therapies designed to limit arginase activity, restore NO availability and reduce oxidative stress. Innovation: This application will, for the firt time, investigate the role of arginase in retinal neurovascular injury. The studies will use molecular approaches to manipulate A1 and A2 expression in combination with real-time vascular imaging, electroretinography and morphometric analyses of neuronal and vascular injury. Therapeutic effects of limiting arginase activity and increasing NO will also be tested. Th research is expected to significantly advance the mechanistic understanding of retinal neurovascular injury and facilitate development of novel strategies for prevention and treatment of ischemic retinopathy.

Public Health Relevance

Lack of blood flow to the retina in diabetic retinopathy and other forms of ischemic retinopathy often leads to blindness. Strong evidence is presented to show that ischemia-induced activity of a particular enzyme pathway initiates a series of events resulting in vascular and neural damage and retinal dysfunction, thereby impairing vision. The proposed studies will define critical steps in the pathological process and test whether novel agents can block this pathway to maintain retinal blood flow and prevent retinal damage and vision loss.

Agency
National Institute of Health (NIH)
Institute
National Eye Institute (NEI)
Type
Research Project (R01)
Project #
2R01EY011766-14A1
Application #
8530914
Study Section
Special Emphasis Panel (ZRG1-BDCN-H (02))
Program Officer
Shen, Grace L
Project Start
1998-03-01
Project End
2018-02-28
Budget Start
2013-03-01
Budget End
2014-02-28
Support Year
14
Fiscal Year
2013
Total Cost
$414,785
Indirect Cost
$138,262
Name
Georgia Regents University
Department
Biology
Type
Schools of Medicine
DUNS #
966668691
City
Augusta
State
GA
Country
United States
Zip Code
30912
Wang, Lin; Bhatta, Anil; Toque, Haroldo A et al. (2015) Arginase inhibition enhances angiogenesis in endothelial cells exposed to hypoxia. Microvasc Res 98:8-Jan
Patel, Chintan; Narayanan, S Priya; Zhang, Wenbo et al. (2014) Activation of the endothelin system mediates pathological angiogenesis during ischemic retinopathy. Am J Pathol 184:3040-51
Narayanan, S P; Xu, Z; Putluri, N et al. (2014) Arginase 2 deficiency reduces hyperoxia-mediated retinal neurodegeneration through the regulation of polyamine metabolism. Cell Death Dis 5:e1075
Suwanpradid, Jutamas; Rojas, Modesto; Behzadian, M Ali et al. (2014) Arginase 2 deficiency prevents oxidative stress and limits hyperoxia-induced retinal vascular degeneration. PLoS One 9:e110604
Jittiporn, Kanjana; Suwanpradid, Jutamas; Patel, Chintan et al. (2014) Anti-angiogenic actions of the mangosteen polyphenolic xanthone derivative ?-mangostin. Microvasc Res 93:72-9
Liu, Hua; Zhang, Wenbo; Xu, Zhimin et al. (2013) Hyperoxia causes regression of vitreous neovascularization by downregulating VEGF/VEGFR2 pathway. Invest Ophthalmol Vis Sci 54:918-31
Yao, Lin; Chandra, Surabhi; Toque, Haroldo A et al. (2013) Prevention of diabetes-induced arginase activation and vascular dysfunction by Rho kinase (ROCK) knockout. Cardiovasc Res 97:509-19
Narayanan, S Priya; Rojas, Modesto; Suwanpradid, Jutamas et al. (2013) Arginase in retinopathy. Prog Retin Eye Res 36:260-80
El-Remessy, Azza B; Franklin, Telina; Ghaley, Nagla et al. (2013) Diabetes-induced superoxide anion and breakdown of the blood-retinal barrier: role of the VEGF/uPAR pathway. PLoS One 8:e71868
Toque, Haroldo A; Nunes, Kenia P; Yao, Lin et al. (2013) Akita spontaneously type 1 diabetic mice exhibit elevated vascular arginase and impaired vascular endothelial and nitrergic function. PLoS One 8:e72277

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