Nanoparticle-mediated expression of a Wnt pathway inhibitor ameliorates ocular neovascularization

Zhongxiao Wang; Rui Cheng; Kyungwon Lee; Puneet Tyagi; Lexi Ding; Uday B Kompella; Jing Chen; Xun Xu; Jian-Xing Ma

doi:10.1161/ATVBAHA.114.304627

Nanoparticle-mediated expression of a Wnt pathway inhibitor ameliorates ocular neovascularization

Arterioscler Thromb Vasc Biol. 2015 Apr;35(4):855-64. doi: 10.1161/ATVBAHA.114.304627. Epub 2015 Feb 5.

Authors

Zhongxiao Wang¹, Rui Cheng¹, Kyungwon Lee¹, Puneet Tyagi¹, Lexi Ding¹, Uday B Kompella¹, Jing Chen¹, Xun Xu², Jian-Xing Ma²

Affiliations

¹ From the Department of Ophthalmology, Shanghai First People's Hospital Affiliated to Shanghai Jiaotong University, Shanghai, China (Z.W., X.X.); Department of Physiology, University of Oklahoma Health Sciences Center, Oklahoma City (Z.W., R.C., K.L., L.D., J.-x.M.); Department of Pharmaceutical Sciences, Skaggs School of Pharmacy and Pharmaceutical Sciences, University of Colorado Denver, Aurora (P.T., U.B.K.); Department of Ophthalmology, Xiangya Hospital, Central South University, Changsha, China (L.D.); and Department of Ophthalmology, Boston Children's Hospital, Harvard Medical School, MA (J.C.).
² From the Department of Ophthalmology, Shanghai First People's Hospital Affiliated to Shanghai Jiaotong University, Shanghai, China (Z.W., X.X.); Department of Physiology, University of Oklahoma Health Sciences Center, Oklahoma City (Z.W., R.C., K.L., L.D., J.-x.M.); Department of Pharmaceutical Sciences, Skaggs School of Pharmacy and Pharmaceutical Sciences, University of Colorado Denver, Aurora (P.T., U.B.K.); Department of Ophthalmology, Xiangya Hospital, Central South University, Changsha, China (L.D.); and Department of Ophthalmology, Boston Children's Hospital, Harvard Medical School, MA (J.C.). jian-xing-ma@ouhsc.edu drxuxun@sjtu.edu.cn.

Abstract

Objective: The deficiency of very low-density lipoprotein receptor resulted in Wnt signaling activation and neovascularization in the retina. The present study sought to determine whether the very low-density lipoprotein receptor extracellular domain (VLN) is responsible for the inhibition of Wnt signaling in ocular tissues.

Approach and results: A plasmid expressing the soluble VLN was encapsulated with poly(lactide-co-glycolide acid) to form VLN nanoparticles (VLN-NP). Nanoparticles containing a plasmid expressing the low-density lipoprotein receptor extracellular domain nanoparticle were used as negative control. MTT, modified Boyden chamber, and Matrigel (™) assays were used to evaluate the inhibitory effect of VLN-NP on Wnt3a-stimulated endothelial cell proliferation, migration, and tube formation. Vldlr(-/-) mice, oxygen-induced retinopathy, and alkali burn-induced corneal neovascularization models were used to evaluate the effect of VLN-NP on ocular neovascularization. Wnt reporter mice (BAT-gal), Western blotting, and luciferase assay were used to evaluate Wnt pathway activity. Our results showed that VLN-NP specifically inhibited Wnt3a-induced endothelial cell proliferation, migration, and tube formation. Intravitreal injection of VLN-NP inhibited abnormal neovascularization in Vldlr(-/-), oxygen-induced retinopathy, and alkali burn-induced corneal neovascularization models, compared with low-density lipoprotein receptor extracellular domain nanoparticle. VLN-NP significantly inhibited the phosphorylation of low-density lipoprotein receptor-related protein 6, the accumulation of β-catenin, and the expression of vascular endothelial growth factor in vivo and in vitro.

Conclusions: Taken together, these results suggest that the soluble VLN is a negative regulator of the Wnt pathway and has antiangiogenic activities. Nanoparticle-mediated expression of VLN may thus represent a novel therapeutic approach to treat pathological ocular angiogenesis and potentially other vascular diseases affected by Wnt signaling.

Keywords: VLDLR; Wnt; eye; nanoparticle; neovascularization.

Publication types

Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't

MeSH terms

Animals
Cell Movement
Cell Proliferation
Cells, Cultured
Cornea / blood supply*
Corneal Neovascularization / genetics
Corneal Neovascularization / metabolism
Corneal Neovascularization / physiopathology
Corneal Neovascularization / prevention & control*
Disease Models, Animal
Humans
Intravitreal Injections
Lactic Acid / chemistry*
Low Density Lipoprotein Receptor-Related Protein-6 / metabolism
Mice, Inbred C57BL
Mice, Knockout
Nanoparticles*
Phosphorylation
Polyglycolic Acid / chemistry*
Polylactic Acid-Polyglycolic Acid Copolymer
RNA Interference
Rats, Sprague-Dawley
Receptors, LDL / genetics
Receptors, LDL / metabolism*
Retinal Neovascularization / genetics
Retinal Neovascularization / metabolism
Retinal Neovascularization / physiopathology
Retinal Neovascularization / prevention & control*
Retinal Vessels / metabolism*
Retinal Vessels / physiopathology
Time Factors
Transfection / methods*
Vascular Endothelial Growth Factor A / metabolism
Wnt Signaling Pathway*
Wnt3A Protein / genetics
Wnt3A Protein / metabolism*
beta Catenin / metabolism

Substances

Low Density Lipoprotein Receptor-Related Protein-6
Receptors, LDL
VLDL receptor
Vascular Endothelial Growth Factor A
Wnt3A Protein
beta Catenin
Polylactic Acid-Polyglycolic Acid Copolymer
Polyglycolic Acid
Lactic Acid

Abstract

Publication types

MeSH terms

Substances

Grants and funding