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)： 由于创伤或受伤造成的角膜疤痕是我们退伍军人和士兵失明的主要原因。它每年影响150万美国人，是全球第三大致盲原因。尽管使用了战斗护目镜，但在阿富汗和伊拉克的战斗中，16%的伤员眼睛受伤。由于缺乏有效的治疗方法，每年在45000名美国人中，20%的伤员被摘除眼球，接受角膜移植。该项目专注于开发基于纳米技术的基因治疗方法，提供长期缓解而没有明显副作用，这是一种治疗角膜失明的新方法。在上一次 在筹资期间，我们在这方面取得了显著进展，这从我们的出版物和奖项中可见一斑。我们成功地确定了一种有效的纳米载体、递送技术和靶向基因治疗方法，并使用兔模型测试了抗转化生长因子�(转化生长因子�)基因在体内治疗角膜失明的潜力。我们的直接目标是测试4个新的假设，形成4个特定的目标：1)测试聚乙烯亚胺结合金纳米颗粒(PEI2-GNPs)在眼睛中的滞留不影响体内角膜的角膜基质细胞和内皮细胞密度或纤维均匀，2)测试 骨形态发生蛋白-7(BMP7)通过PEI2-GNP在角膜中传递，通过减轻纤维化的SmAD信号，增加分化蛋白的抑制物，阻断角膜角质细胞向肌成纤维细胞的分化，调节角膜愈合，这是体内瘢痕形成的机制，3)试验表明，由肝细胞生长因子和骨形态发生蛋白7或可溶性转化生长因子�受体II(sTGF�RII)和骨形态发生蛋白7基因组成的2基因联合治疗是一种创新的方法，可以最大限度地解决体内固定和愈合的角膜瘢痕，4)实验表明，通过眼部局部应用涡流调节剂(每天两次，连续3天)，对转化生长因子�诱导的促纤维化基因进行广泛的表观遗传学敲除是治疗体内角膜瘢痕形成的一种新方法。 没有严重的副作用。我们已发表和正在进行的研究有力地支持了这些假设。成人眼的角膜透明和正常功能需要22,000-24000个角膜细胞/毫米~3，2,400-3,200个内皮细胞/毫米~2和特征性的胶原纤维组织。我们正在进行的研究表明，pEI2-GNPs是一种安全有效的眼部给药载体，转导转化生长因子�RII基因抑制转化生长因子�，骨形态发生蛋白7调控SmAD途径，肝细胞生长因子选择性地诱导角膜肌成纤维细胞凋亡。我们的实验室拥有新颖的活体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、Western blotting和分子生物学技术方面拥有丰富的经验。这项研究的成功完成将大大提高我们对角膜创伤愈合分子机制的理解，继续/推进新型眼部给药系统和新的角膜失明治疗方法的开发，并推动视力研究领域的发展。