Gene therapy in the cornea

角膜基因治疗

基本信息

批准号：
7882845
负责人：
Rajiv Ravindra Mohan
金额：
$ 10.04万
依托单位：
UNIVERSITY OF MISSOURI-COLUMBIA
依托单位国家：
美国
项目类别：
财政年份：
2007
资助国家：
美国
起止时间：
2007-04-01 至 2012-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7882845
关键词：
Actins Adult Adverse effects Affect Age American Animal Disease Models Animal Model Appearance Blindness Cells Chickens Clinical Competence Cornea Corneal Diseases Corneal Neovascularization Corneal dystrophy Cytomegalovirus Defect Disease Experimental Models Fibrosis Functional disorder Gene Delivery Gene Transfer Genes Genetic Goals Human Hybrids In Vitro Infection Injury Kidney Knockout Mice Length Leucine Ligaments Location Lung Methods Modeling Mus Myofibroblast Organ Oryctolagus cuniculus Pathogenesis Plasmid Cloning Vector Proteoglycan Reporting Role Serotyping Site Subfamily lentivirinae Surgical Flaps TGFBI gene Techniques Testing Time Tissues Transgenes Transgenic Animals Transgenic Organisms angiogenesis decorin dosage gene function gene therapy in vivo mouse model neovascularization novel novel strategies prevent promoter research study selective expression targeted delivery therapeutic gene transgene expression vector

项目摘要

DESCRIPTION: Corneal haze and neovascularization affect over 1.5 million Americans every year and are among the leading causes of blindness worldwide. Gene therapy is an attractive and novel approach to prevent/treat these corneal disorders. However, clinical utility of gene therapy is severely limited due to unavailability of tissue-targeted gene transfer methods. To develop tissue-targeted selective gene therapy approaches for the cornea, we hypothesized that foreign genes can be selectively expressed in the cornea at the desired site for selected duration using appropriate vectors and vector-delivery techniques. Preliminary in vivo experiments performed with AAV serotype 2 or 5, lentivirus or plasmid vectors containing CMV or hybrid CMV+chicken-¿-actin promoter, and defined vector-delivery techniques demonstrated that transgene can be precisely expressed in keratocytes of normal/damaged corneas in vivo for the desired time period. Decorin (a small leucine-rich proteoglycan) gene therapy has been shown to prevent fibrosis and angiogenesis in various disease animal models. These reports led us to hypothesize that selective expression of decorin in keratocytes can inhibit/prevent corneal haze and neovascularization with minimal side effects. The in vitro studies performed to test this hypothesis demonstrated competence of decorin to inhibit keratocyte transformation to myofibroblasts. This transformation is known to cause corneal haze in vivo. We further hypothesize that selective tissue-targeted gene transfer approaches can be used to develop animal models for studying the specific function of disease-causing genes such as TGF¿ and BIGH3 in the adult cornea in vivo without altering their expression in vital organs. The function of such genes cannot be studied using conventional transgenic approaches because TGF¿-deficient transgenic animals suffer lethal defects and die by 4 weeks of age. The specific aims to test the hypotheses are 1) vector and vector-delivery techniques regulate level, duration, and location of transgene expression in keratocytes in vivo 2) decorin gene therapy can control corneal haze and 3) decorin gene therapy can inhibit corneal neovascularization. Using a mouse model, tested vectors, and optimized vector delivery-techniques, we will test Specific Aim 1 and will define short- and long-term selective gene transfer approaches for the cornea. Rabbit models will be used to test Specific Aims 2 and 3 by delivering decorin into keratocytes with optimal selective gene transfer methods.

描述：角膜阴霾和新血管化每年影响超过150万美国人，并且是全球失明的主要原因之一。基因疗法是预防/治疗这些角膜疾病的一种有吸引力且新颖的方法。然而，由于靶向组织靶向基因转移方法，基因治疗的临床实用性受到严重限制。为了开发有关角膜的靶向组织的选择性基因治疗方法，我们假设可以使用适当的矢量和矢量分解技术在所需的持续时间在所需位置的角膜中选择外源基因。使用AAV血清型2或5，含有CMV或杂化CMV+CMV+Chicken-actin启动子的慢病毒或质粒载体进行的初步体内实验，并定义了载体 - 分发技术，证明可以在正常/损坏的Corneras desired Times vivo中准确表达转换。 Decorin（一种小亮氨酸蛋白聚糖）的基因疗法已被证明可预防各种疾病动物模型中的纤维化和血管生成。这些报道使我们假设在角膜细胞中的选择性表达可以抑制/防止角膜雾和新生血管形成，而副作用最少。对该假设进行检验的体外研究表明，装饰蛋白抑制角膜细胞转化为肌纤维细胞的能力。已知这种转化在体内引起角膜阴霾。我们进一步假设，可以使用选择性组织为靶向组织的基因转移方法来开发动物模型，以研究引起疾病的基因的特定功能，例如TGF？和BIGH3在体内成人角膜中，而不会改变其在重要器官中的表达。这种基因的功能无法使用常规转基因方法研究，因为TGF¿-缺乏的转基因动物患有致命的缺陷，并在4周龄时死亡。测试假设的具体目的是1）载体，载体分娩技术调节了体内角化核细胞中转化表达的水平，持续时间和位置2）DecorIn基因疗法可以控制角膜雾性，3）Decorin Gene疗法可以抑制角膜新的血管性新血管生成。使用小鼠模型，测试的向量和优化的矢量输送技术，我们将测试特定的目标1，并将定义角膜的短期和长期选择性基因转移方法。兔模型将通过使用最佳选择性基因转移方法将Decorin传递到角膜细胞中来测试特定目标2和3。