The overall goal of this project is to test the hypothesis that increased lysyl oxidase (LOX) expression and activation triggered by high glucose/diabetes compromises basement membrane (BM) function and thereby induces excess permeability and vascular cell death in diabetic retinopathy (DR). The hypothesis is based on findings that diabetes increases expression of LOX, which is required for the formation and maturation of the BM, and that HG-induced LOX upregulation and over-activation compromises endothelial barrier function. Importantly, inhibition of HG-induced LOX overexpression results in reduced cell monolayer permeability. While LOX is known for its cross-linking activity, LOX propeptide (LOX-PP), released during LOX biogenesis, is known for regulating cell proliferation and survival through modulation of Akt and Erk pathways. Our preliminary data indicates that HG-induced LOX overexpression promotes apoptosis in retinal endothelial cells, and that the process may be exacerbated by LOX-PP, which is upregulated by HG. Furthermore, preliminary data indicates that LOX binds with high affinity to extracellular fibronectin (FN) under HG condition, and interferes with LOX internalization into the cell, a process unique to LOX. Thus, the rationale for the proposed studies is that (i) HG-induced LOX overexpression contributes to capillary leakage, and together with increased LOX-PP promotes apoptosis by modulating Akt and Erk pathways, and (ii) HG-induced increased binding of LOX with ECM proteins, decreases LOX internalization, attenuates negative feedback signaling and thereby promotes LOX overexpression. The proposed studies will be performed using biochemical, molecular, and genetic approaches utilizing cell culture models, diabetic rats, and LOX heterozygous KO mice (LOX+/-). Towards this goal, we have generated LOX+/- mice colonies at our animal facility that are now available for the proposed studies.
The specific aims of this proposal are to determine (i) whether inhibition of LOX overexpression and activation prevents apoptosis in retinal endothelial cells in vitro, and reduces vascular cell loss and permeability in diabetic rat retinas (ii) whether the decreased LOX level in LOX+/- mice is protective against diabetes-induced retinal vascular lesions, and (iii) mechanism(s) by which HG induces LOX overexpression and promotes retinal vascular cell loss. Findings from these studies are expected to identify a novel therapeutic strategy for preventing retinal vascular cell loss and capillary leakage, and provide mechanistic insights underlying HG-induced LOX overexpression in DR.

Public Health Relevance

The rising incidence and prevalence of diabetes worldwide has resulted in a dramatic increase in the occurrence of diabetic retinopathy, a major cause of blindness in the working age population. Current treatment modalities do not cure diabetic retinopathy; therefore, it remains a significant public health issue. Retinal capillary leakage is serious clinical issue in patients with DR. The proposed research aims at preventing vascular permeability by reducing abnormal LOX overexpression in the diabetic retina. If successful, this could lead to the development of a new and effective strategy to treat patients with diabetic retinopathy.

Agency
National Institute of Health (NIH)
Institute
National Eye Institute (NEI)
Type
Research Project (R01)
Project #
5R01EY025528-03
Application #
9248395
Study Section
Diseases and Pathophysiology of the Visual System Study Section (DPVS)
Program Officer
Shen, Grace L
Project Start
2015-04-01
Project End
2020-03-31
Budget Start
2017-04-01
Budget End
2018-03-31
Support Year
3
Fiscal Year
2017
Total Cost
$391,500
Indirect Cost
$166,500
Name
Boston Medical Center
Department
Type
Independent Hospitals
DUNS #
005492160
City
Boston
State
MA
Country
United States
Zip Code
02118
Song, Brian; Kim, Dongjoon; Nguyen, Ngan-Ha et al. (2018) Inhibition of Diabetes-Induced Lysyl Oxidase Overexpression Prevents Retinal Vascular Lesions Associated With Diabetic Retinopathy. Invest Ophthalmol Vis Sci 59:5965-5972
Roy, Sayon; Jiang, Jean X; Li, An-Fei et al. (2017) Connexin channel and its role in diabetic retinopathy. Prog Retin Eye Res 61:35-59
Roy, Sayon; Kim, Dongjoon; Lim, Remington (2017) Cell-cell communication in diabetic retinopathy. Vision Res 139:115-122
Roy, Sayon; Kern, Timothy S; Song, Brian et al. (2017) Mechanistic Insights into Pathological Changes in the Diabetic Retina: Implications for Targeting Diabetic Retinopathy. Am J Pathol 187:9-19
Roy, Sumon; Kim, Dongjoon; Hernández, Cristina et al. (2015) Beneficial effects of fenofibric acid on overexpression of extracellular matrix components, COX-2, and impairment of endothelial permeability associated with diabetic retinopathy. Exp Eye Res 140:124-129