纳米颗粒介导的Wnt信号通路抑制剂表达可改善眼部新生血管形成。

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

作者信息

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

机构信息

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.).

出版信息

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

DOI:10.1161/ATVBAHA.114.304627

PMID:25657312

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC4376607/

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.

摘要

目的

极低密度脂蛋白受体的缺乏导致视网膜中Wnt信号激活和新生血管形成。本研究旨在确定极低密度脂蛋白受体细胞外结构域（VLN）是否负责抑制眼组织中的Wnt信号。

方法与结果

将表达可溶性VLN的质粒用聚（丙交酯-共-乙交酯酸）包裹以形成VLN纳米颗粒（VLN-NP）。含有表达低密度脂蛋白受体细胞外结构域纳米颗粒的质粒的纳米颗粒用作阴性对照。采用MTT法、改良博伊登小室法和基质胶（™）实验来评估VLN-NP对Wnt3a刺激的内皮细胞增殖、迁移和管腔形成的抑制作用。利用Vldlr（-/-）小鼠、氧诱导性视网膜病变和碱烧伤诱导的角膜新生血管形成模型来评估VLN-NP对眼新生血管形成的作用。采用Wnt报告基因小鼠（BAT-gal）、蛋白质免疫印迹法和荧光素酶实验来评估Wnt信号通路活性。我们的结果表明，VLN-NP特异性抑制Wnt3a诱导的内皮细胞增殖、迁移和管腔形成。与低密度脂蛋白受体细胞外结构域纳米颗粒相比，玻璃体内注射VLN-NP可抑制Vldlr（-/-）、氧诱导性视网膜病变和碱烧伤诱导的角膜新生血管形成模型中的异常新生血管形成。VLN-NP在体内和体外均显著抑制低密度脂蛋白受体相关蛋白6的磷酸化、β-连环蛋白的积累以及血管内皮生长因子的表达。