Suprachoroidal nonviral gene transfer of engineered VEGF antagonists

工程 VEGF 拮抗剂的脉络膜上非病毒基因转移

基本信息

批准号：
10557124
负责人：
Peter A Campochiaro
金额：
$ 59.89万
依托单位：
JOHNS HOPKINS UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-02-01 至 2025-01-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10557124
关键词：
Address Adult Affinity Age related macular degeneration Ambulatory Care Facilities Angiogenesis Inhibitors Angiogenic Proteins Animal Model Binding Binding Proteins Biocompatible Materials Biomedical Engineering Biotechnology Blindness Carrying Capacities Cataract Cell Nucleus Cells Characteristics Chronic Complementary DNA DNA DNA Binding DNA cassette Development Disadvantaged Disease Disease model Dose Endocytosis Endosomes Engineering Enzyme-Linked Immunosorbent Assay Esters Extravasation Eye diseases Family Family member Formulation Gene Delivery Gene Transfer Genes Genetic Growth Factor Growth Factor Gene Immune response In Vitro Inflammation Injections Innate Immune Response Intervention Lasers Ligands Liquid substance Measures Medical Modeling Molecular Mutation Nanotechnology Nuclear Nucleic Acids Operating Rooms Ophthalmology Oryctolagus cuniculus Outcome Patients Performance Photoreceptors Plasmids Polymer Chemistry Polymers Positioning Attribute Protein Engineering Proteins Rattus Retina Retinal Degeneration Retinal Detachment Retinal Diseases Risk Route Safety Sclera Site Specificity Structure Structure of retinal pigment epithelium Technology Testing Therapeutic Time Transfection Transgenic Mice Treatment Efficacy United States VEGF165 VEGFA gene Vascular Diseases Vascular Endothelial Growth Factors Vertebral column Viral Vector Vision Vitrectomy adaptive immune response angiogenesis antagonist antiangiogenesis therapy biodegradable polymer cell type clinical care comparative efficacy design diabetic experience experimental study gene replacement gene therapy high risk immunogenic improved in vivo inherited retinal degeneration macular edema manufacture mutant nanoparticle nanopolymer next generation non-viral gene delivery novel novel strategies novel therapeutics promoter protein expression retinal damage standard of care subretinal injection success tertiary amine therapeutic protein therapeutic transgene tool transduction efficiency transgene expression uptake vector vector induced

项目摘要

Project Summary Ocular gene therapy has great potential for sustained expression of anti-angiogenic proteins for retinal/choroidal vascular diseases and also for replacement of mutant genes in patients with inherited retinal degenerations. Viral vectors provide efficient transduction of photoreceptors and retinal pigmented epithelial (RPE) cells when injected into the subretinal space, but not when injected into the vitreous cavity. Subretinal injections require vitrectomy, which carries small but significant risks of vision-threatening complications. Viral vectors induce an immune response which can cause bystander damage to retinal cells and potentially reduce the level and duration of transgene expression. Nonviral gene transfer is less immunogenic and also enables a large carrying capacity for genetic cargos, but has been limited by inefficient transfection. We have developed a new approach, suprachoroidal nonviral gene transfer, which provides widespread, prolonged, robust transgene expression in photoreceptors and RPE cells after suprachoroidal injection of engineered biodegradable polymeric nanoparticles. We propose to optimize the polymer structure and nanoparticle formulation, expression plasmid characteristics, and promoter to maximize the level, topographical extent, and duration of transgene expression in the retina and RPE cells. Suprachoroidal injections are performed in an outpatient clinic setting and can be safer than subretinal injections. The nanoparticles will be used for nonviral suprachoroidal gene transfer of vascular endothelial growth factor (VEGF)165, to determine if effects are comparable to those seen with high level expression of VEGF165 in transgenic mice and to establish new models of retinal/choroidal vascular disease in rats and rabbits. In parallel, we will create a new bioengineered protein that blocks angiogenesis by antagonizing a broad range of VEGF family ligands with high affinity and that is expressed at high levels by the newly engineered vector. After suprachoroidal injection of our new nonviral anti-VEGF vector, we will measure protein expression level, duration of expression, and therapeutic efficacy in established models of retinal/choroidal vascular disease and in the new disease models that we develop. These studies address a major unmet need in highly prevalent eye diseases by developing a noninvasive approach to achieve robust, long term suppression of VEGF family members. Further, the creation of an enabling nonviral gene delivery technology and a novel therapeutic anti-angiogenesis protein could be applicable to many other diseases as well.

项目摘要眼基因治疗具有持续表达抗血管生成蛋白的巨大潜力视网膜/脉络膜血管疾病以及用于遗传性视网膜患者的突变基因的替代退化。病毒载体可有效地转导光感受器和视网膜色素上皮（RPE）将细胞注入视网膜下空间，但在注射到玻璃体腔中时不会。视网膜下注射需要玻璃体切除术，这带来了威胁性并发症的小但重大风险。病毒性的向量引起免疫反应，该反应可能导致旁观者对视网膜细胞的损害，并有可能减少转基因表达的水平和持续时间。非病毒基因转移的免疫原性较少，也可以实现遗传性陈代的较大承载能力，但受到效率低下的转染的限制。我们已经发展了一种新的方法，即螺节非病毒基因转移，可提供广泛，延长，稳定的在跨齿状体注射后，光感受器和RPE细胞中的转基因表达可生物降解的聚合纳米颗粒。我们建议优化聚合物结构和纳米颗粒配方，表达质粒特征和启动子，以最大程度地提高水平，地形范围和视网膜和RPE细胞中转基因表达的持续时间。在门诊诊所设置，并且可以比下视网膜下注射更安全。纳米颗粒将用于非病毒血管内皮生长因子（VEGF）165的上齿状基因转移，以确定效果是否为与转基因小鼠中VEGF165高水平表达的人相媲美，并建立新的大鼠和兔子中视网膜/脉络膜血管疾病的模型。同时，我们将创建一个新的生物工程通过拮抗具有高亲和力和这是新工程矢量在高水平上表达的。在注射我们的新的非病毒抗VEGF载体，我们将测量蛋白质表达水平，表达持续时间和治疗性在视网膜/脉络膜血管疾病的既定模型以及我们的新疾病模型中的功效发展。这些研究通过开发一个高度普遍的眼科疾病的主要需求无创的方法可以实现对VEGF家庭成员的坚固，长期抑制。此外，创建启用非病毒基因递送技术和新型的治疗性抗血管生成蛋白也可能适用于许多其他疾病。