Targeted Gene Therapy and Nanomedicine Approaches to Treat Corneal Diseases

靶向基因疗法和纳米医学方法治疗角膜疾病

• 批准号: 9280757
• 负责人: Rajiv Ravindra Mohan
• 金额: --
• 依托单位: HARRY S. TRUMAN MEMORIAL VA HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-04-01 至 2017-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9280757
• 关键词: Adult; Adverse effects; Affect; Afghanistan; American; Animals; Apoptosis; Apoptotic; Award; Basic Science; Binding; Biological; Blindness; Caring; Cells; Characteristics; Cicatrix; Clinical; Collagen; Collagen Fibril; Combined Modality Therapy; Confocal Microscopy; Cornea; Corneal Diseases; Corneal Injury; Data; Development; Drug Delivery Systems; Electron Microscopy; Endothelium; Epigenetic Process; Excision; Extracellular Matrix; Eye; Eye Injuries; FDA approved; Fibrosis; Funding; Gene Expression; Gene Targeting; Gene Transfer; Genes; Goals; Goggles; Gold; HGF gene; Health; Histology; Human; Hybrids; Image; Impotence; In Vitro; Infection; Inhibitor of Differentiation Proteins; Injury; Innovative Therapy; Iraq; Keratoplasty; Knowledge; Lead; Manuscripts; Mediating; Methods; Military Personnel; Mission; Mitotic; Modality; Modeling; Molecular; Molecular Biology Techniques; Myofibroblast; Nanotechnology; Oryctolagus cuniculus; Pathologic; Pathway interactions; Pharmaceutical Preparations; Phenotype; Pilot Projects; Polyethyleneimine; Production; Property; Protocols documentation; Publications; Publishing; Regulation; Reporting; Research; Research Design; Resolution; Role; Signal Transduction; Smad7 protein; System; Techniques; Testing; Toxic effect; Transcription Repressor/Corepressor; Transforming Growth Factors; Translational Research; Trauma; Veterans; Vision research; Vorinostat; War; Western Blotting; Wound Healing; base; bone morphogenic protein; combat; combination gene therapy; corneal scar; density; experience; extracellular; fighting; gene therapy; healing; health administration; improved; in vivo; injured; innovation; knock-down; light scattering; nanomedicine; nanoparticle; non-viral gene therapy; novel; novel strategies; novel therapeutics; pre-clinical research; prevent; public health relevance; receptor; residence; transcription factor; transgene expression; vector

DESCRIPTION (provided by applicant): Corneal scarring due to trauma or injury is a major cause of blindness in our veterans and troops. It affects 1.5 million Americans each year and is the 3rd leading cause of global blindness. Despite using combat goggles, eye injuries occurred in 16% of wounded troops fighting in Afghanistan and Iraq. A lack of efficient therapies caused eye removal in 20% of injured troops and corneal transplantation in >45,000 American each year. This project focuses on developing nanotechnology-based gene therapy approaches that offer long-term relief without significant side effects, which is a novel modality to treat corneal blindness. During last funding period, we made remarkable progress to this end as evident from our publications and awards. We successfully identified a potent nanoparticle vector, delivery techniques, and targeted gene therapy approaches, and we tested the potential of anti-transforming growth factor-� (TGF�) genes to treat corneal blindness in vivo using a rabbit model. Our immediate goal is to test 4 novel hypotheses formulated into 4 specific aims: 1) test that the residence of polyethyleneimine conjugated gold nanoparticles (PEI2-GNPs) in the eye does not affect corneal keratocyte and endothelial density or fibril uniformity in the cornea in vivo, 2) test that bone morphogenic protein-7 (BMP7) delivery in the cornea via PEI2-GNP regulates corneal healing by mitigating fibrotic Smad signaling, increasing inhibitors of differentiation proteins, ad blocking corneal keratocyte differentiation to myofibroblasts, a mechanism of scar development in vivo, 3) test that 2-gene combination therapy consisting of hepatocyte growth factor (HGF) and BMP7 or soluble TGF� receptor II (sTGF�RII) and BMP7 genes is an innovative approach to achieve maximum resolution of fixed- and healing- corneal scars in vivo, and 4) test that a broad-based epigenetic knockdown of TGF�-induced profibrotic genes by a topical vorinostat (twice a day for 3 days) application on the eye is a novel method to treat corneal scarring in vivo without major side effects. Our published and ongoing studies strongly support these hypotheses. For corneal clarity and normal function in the adult human eye, 22,000-24000 keratocytes/mm3, 2400-3200 endothelial/mm2 and characteristic collagen fibril organization are necessary. Our ongoing studies suggest that PEI2-GNPs are a safe and potent vector for ocular drug delivery, sTGF�RII gene transfer inhibits TGF�, BMP7 modulates the Smad pathway, and HGF selectively induces apoptosis in corneal myofibroblasts. Our lab has novel in vivo rabbit and in vitro human corneal fibrosis models and substantial experience in performing animal studies, clinical eye imaging, nanoparticle characterization, histology, electron microscopy, western blotting and molecular biology techniques. Successful completion of the proposed research will significantly improve our understanding of the molecular mechanisms of corneal wound healing, continue/progress the development of a novel ocular delivery system and newer therapies for corneal blindness, and advance the vision research field.

描述（由申请人提供）： 外伤或受伤造成的角膜疤痕是退伍军人和军队失明的主要原因，它每年影响 150 万美国人，是全球失明的第三大原因，尽管使用了战斗护目镜，但仍有 16% 的受伤军人发生眼部受伤。在阿富汗和伊拉克的战斗中，由于缺乏有效的治疗方法，每年有 20% 的受伤士兵被摘除眼睛，并导致超过 45,000 名美国人接受角膜移植。该项目致力于开发基于纳米技术的基因治疗方法。长期缓解且无明显副作用，是治疗角膜失明的新方法。 在资助期间，我们在这方面取得了显着进展，从我们的出版物和奖项中可以看出，我们成功地确定了一种有效的纳米颗粒载体、递送技术和靶向基因治疗方法，并且我们测试了抗转化生长因子-�（TGF）的潜力。 �) 使用兔子模型在体内治疗角膜失明的基因，我们的直接目标是测试 4 个新假设，这些假设被制定为 4 个具体目标：1) 测试聚乙烯亚胺缀合金纳米粒子 (PEI2-GNP) 的驻留。眼内不影响角膜体内角膜细胞和内皮细胞密度或原纤维均匀性，2) 测试 通过 PEI2-GNP 在角膜中递送骨形态发生蛋白 7 (BMP7)，通过减轻纤维化 Smad 信号传导、增加分化蛋白抑制剂、阻止角膜角质细胞分化为肌成纤维细胞（体内疤痕形成的机制）来调节角膜愈合，3) 测试由肝细胞生长因子 (HGF) 和 BMP7 或可溶性 TGFα 受体 II (sTGFαRII) 组成的 2 基因联合疗法，以及BMP7 基因是一种创新方法，可在体内实现固定和愈合角膜疤痕的最大分辨率，4) 测试局部伏立诺他（每天两次，持续 3 天）对 TGFβ 诱导的促纤维化基因的广泛表观遗传敲低）应用于眼睛是一种体内治疗角膜疤痕的新方法 我们已发表和正在进行的研究强烈支持这些假设，对于成年人眼的角膜清晰度和正常功能来说，22,000-24000 个角膜细胞/mm3、2400-3200 个内皮细胞/mm2 和特征性胶原纤维组织是必要的。研究表明 PEI2-GNP 是一种安全有效的眼部药物递送载体，sTGF�RII 基因转移抑制 TGF�， BMP7 调节 Smad 通路，HGF 选择性诱导角膜肌成纤维细胞凋亡。我们的实验室拥有新颖的体内兔和体外人角膜纤维化模型，并在动物研究、临床眼成像、纳米颗粒表征、组织学、电子显微镜、蛋白质印迹方面拥有丰富的经验。印迹和分子生物学技术的成功完成将显着提高我们对角膜伤口愈合分子机制的理解，继续/推进新型眼部输送系统的开发。以及针对角膜失明的新疗法，并推进视力研究领域。