Our recent studies have demonstrated that abnormal activation of the Wnt pathway plays key roles in pathogenesis of retinal inflammation, NV and fibrosis in both AMD and diabetic retinopathy. A genetic study has reported that variants of very low-density lipoprotein receptor (VLDLR) are associated with AMD. We have recently reported that VLDLR functions as a negative regulator of the Wnt pathway, as VLDLR knockout resulted in Wnt pathway over-activation, leading to AMD-like pathologies, such as retinal inflammation, vascular leakage and sub-retinal NV. Toward the mechanism by which VLDLR regulates the Wnt pathway, we have recently obtained the following preliminary data: 1) Expression of VLDLR and its soluble extracellular domain (VLDLRN) inhibits Wnt signaling. 2) The VLDLRN peptide decreases total LRP6 protein levels. 3) Co-immunoprecipitation assay showed that VLDLRN binds with LRP6, forming a VLDLR-LRP6 heterodimer. 4) VLDLRN peptide blocks Fz-LRP6 dimerization induced by Wnt ligand. Based on these observations, we hypothesize that VLDLR forms a heterodimer with LRP6, which blocks the Wnt ligand-induced LRP6 aggregation and LRP6 signalosome formation. This binding may represent a mechanism by which VLDLR inhibits Wnt signaling. To test this hypothesis, we will determine if binding of VLDLR to LRP6 blocks the Wnt ligand-induced LRP6 aggregation and formation of signalosomes. We will also determine if binding of VLDLR to LRP6 affects LRP6 endocytosis and stability. Further, we will define the sequence domains responsible for the interaction between VLDLR and LRP6 using deletion mutants and co-immunoprecipitation assays. The therapeutic potential of the sequence domain of VLDLR binding to LRP6 and inhibiting Wnt signaling will be explored by evaluating its efficacy on retinal inflammation, vascular leakage and NV. Recently, two independent large clinical trials reported that fenofibrate, a PPAR? agonist which lowers VLDL levels in the circulation, has therapeutic effects on retinal vascular leakage and NV in type 2 diabetic patients. Our preliminary studies found that fenofibrate inhibits Wnt signaling and up- regulates VLDLR expression and its promoter activity. Therefore, we will test the hypothesis that inhibition of Wnt signaling through up-regulation of VLDLR expression by fenofibrate represents a mechanism for its beneficial effects on retinal inflammation, vascular leakage and NV. We will use VLDLR KO mice, PPAR? KO mice and primary RPE and endothelial cells from these KO mice to determine if VLDLR and PPAR? are essential for mediating the Wnt-inhibiting effect of fenofibrate. These studies will elucidate the mechanism by which VLDLR regulates the Wnt pathway and identify a novel, endogenous regulatory mechanism for this important pathway. The proposed studies will reveal the interactions between PPAR? and the canonical Wnt pathway and contribute to the development of new treatment for AMD.

Public Health Relevance

Choroidal neovascularization is a severe complication of age-related macular degeneration (AMD) and a major cause of blindness in aged population. This project aims to explore a novel pathogenic mechanism, i.e, activation of the wnt signal pathway, for choroidal neovascularization. This project may also identify a clinical drug for the treatment of AMD.

Agency
National Institute of Health (NIH)
Institute
National Eye Institute (NEI)
Type
Research Project (R01)
Project #
5R01EY018659-05
Application #
8517122
Study Section
Biology and Diseases of the Posterior Eye Study Section (BDPE)
Program Officer
Shen, Grace L
Project Start
2007-12-01
Project End
2014-07-31
Budget Start
2013-08-01
Budget End
2014-07-31
Support Year
5
Fiscal Year
2013
Total Cost
$448,496
Indirect Cost
$145,458
Name
University of Oklahoma Health Sciences Center
Department
Physiology
Type
Schools of Medicine
DUNS #
878648294
City
Oklahoma City
State
OK
Country
United States
Zip Code
73117
Hu, Shuqun; Liu, Hua; Ha, Yonju et al. (2015) Posttranslational modification of Sirt6 activity by peroxynitrite. Free Radic Biol Med 79:176-85
McBride, Jeffrey D; Jenkins, Alicia J; Liu, Xiaochen et al. (2014) Elevated circulation levels of an antiangiogenic SERPIN in patients with diabetic microvascular complications impair wound healing through suppression of Wnt signaling. J Invest Dermatol 134:1725-34
Zhou, Kelu Kevin; Benyajati, Siribhinya; Le, Yun et al. (2014) Interruption of Wnt signaling in Müller cells ameliorates ischemia-induced retinal neovascularization. PLoS One 9:e108454
Moran, Elizabeth; Ding, Lexi; Wang, Zhongxiao et al. (2014) Protective and antioxidant effects of PPAR? in the ischemic retina. Invest Ophthalmol Vis Sci 55:4568-76
Takahashi, Yusuke; Moiseyev, Gennadiy; Ma, Jian-xing (2014) Identification of key residues determining isomerohydrolase activity of human RPE65. J Biol Chem 289:26743-51
Lee, Kyungwon; Shin, Younghwa; Cheng, Rui et al. (2014) Receptor heterodimerization as a novel mechanism for the regulation of Wnt/?-catenin signaling. J Cell Sci 127:4857-69
Qiu, F; He, J; Zhou, Y et al. (2014) Plasma and vitreous fluid levels of Dickkopf-1 in patients with diabetic retinopathy. Eye (Lond) 28:402-9
Ding, Lexi; Cheng, Rui; Hu, Yang et al. (2014) Peroxisome proliferator-activated receptor ? protects capillary pericytes in the retina. Am J Pathol 184:2709-20
Wang, Zhongxiao; Moran, Elizabeth; Ding, Lexi et al. (2014) PPAR? regulates mobilization and homing of endothelial progenitor cells through the HIF-1?/SDF-1 pathway. Invest Ophthalmol Vis Sci 55:3820-32
Hu, Yang; Chen, Ying; Lin, Mingkai et al. (2013) Pathogenic role of the Wnt signaling pathway activation in laser-induced choroidal neovascularization. Invest Ophthalmol Vis Sci 54:141-54

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