DNA Delivery for Treatment of Proliferative Vitreoretinopathy and Ocular Scarring

DNA 递送治疗增殖性玻璃体视网膜病变和眼部疤痕

• 批准号: 7761662
• 负责人: Horst A. von Recum
• 金额: $ 19.43万
• 依托单位: CASE WESTERN RESERVE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-02-01 至 2012-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7761662
• 关键词: Address; Age related macular degeneration; Antisense Oligonucleotides; Apoptosis; Area; Biological; Biological Assay; Cataract; Cataract Extraction; Cell Culture Techniques; Cell Line; Cell Survival; Cell model; Cells; Cicatrix; Clinic; Clinical; Code; Cornea; DNA; DNA delivery; Data; Dendrimers; Diabetic Retinopathy; Disease; Drug Delivery Systems; Drug Formulations; Encapsulated; Environment; Eye; Eye diseases; Familiarity; Fiber; Fibroblasts; Fluorescence; Fluorescence Microscopy; Future; Gene Expression; Gene Silencing; Glaucoma; Goals; Green Fluorescent Proteins; Half-Life; Human; Implant; Investigation; Kinetics; Laboratories; Lead; Liposomes; Liquid substance; Macular degeneration; Micelles; Modeling; Nucleic Acids; Oligonucleotides; Operative Surgical Procedures; Outcome; Patient Care; Peptides; Pharmaceutical Preparations; Phase; Polymers; Predisposition; Process; Proliferative Vitreoretinopathy; Public Health; Research; Research Personnel; Simulate; Small Interfering RNA; Solutions; Staining method; Stains; Surface; System; Testing; Therapeutic; Therapeutic Effect; Time; Tissues; Transfection; Transforming Growth Factor beta; Translations; Up-Regulation; Vascular Endothelial Growth Factors; Western Blotting; Work; annexin A5; aptamer; aqueous; base; biodegradable polymer; compliance behavior; controlled release; corneal scar; cost; effective therapy; glaucoma surgery; in vivo; innovation; knock-down; nanofiber; nanoscale; nanosized; novel; novel therapeutics; overexpression; pressure; protein expression; public health relevance; scaffold; small molecule; success; vector

DESCRIPTION (provided by applicant): While many new therapeutic molecules for treatment of ocular diseases have become available over the past decade, one of the major problems facing their effective application in clinic has been drug delivery. Often bioactive molecules have a short half-life in vivo and require repeated administration. In the case of intraocular administration this often means poor patient compliance and high cost. As such, controlled release of therapeutic molecules over a period of months to years is expected to have a significant impact in patient care and in the treatment of ocular disease. The focus of this proposal is to investigate micro and nanoscale polymer fibers for the intraocular delivery of therapeutic molecules. Fibers have an advantage over other nano and microscale delivery vectors in that they can be moved or removed if complications arise. One of the primary targets implicated in several ocular diseases is Transforming Growth Factor - beta (TGF-¿). These diseases include corneal scarring, scarring following cataract and glaucoma surgery, as well as proliferative vitreoretinopathy. Decreasing TGF-¿ expression and its effect through use of antisense oligonucleotides, aptamers and other small molecule drugs has shown some success in treatment of these diseases. While delivery of siRNA against TGF-¿ has also been demonstrated to be effective, siRNA is limited as a long term therapy due to its susceptibility to ubiquitous RNAses as well as the need to have a large number of molecules for effective knockdown in vivo. In this study we propose to examine delivery of DNA which codes for the siRNA to knockdown TGF-¿ expression. Delivery of DNA addresses issues of stability since DNA is much more stable than RNA, and addresses issues of drug amount since a single DNA molecule can be transcribed into many siRNAs. The objective of the proposed research is to develop a microscale drug delivery platform which can easily be implanted to treat a wide range of ocular diseases. Specifically the hypothesis of this study is that the proposed therapeutic DNA can be incorporated and released from polymer nanofibers, and that released DNA can result in expression knockdown in ocular cell models. The hypothesis will be tested in three specific aims: 1) Evaluate the bioactivity of DNA released from nanofibers formed by electrospinning, 2) In a destabilized GFP model evaluate effect of released DNA in knocking down expression, and 3) In a human corneal fibroblast model evaluate the effect of released DNA in knockdown of TGF-¿ expression. This ocular model was chosen due to the investigators' familiarity with it. However, a drug delivery platform which can successfully deliver therapeutic DNA against one disease could rapidly be applied toward treatment of other diseases, such as VEGF knockdown in wet-type macular degeneration, or in diabetic retinopathy. PUBLIC HEALTH RELEVANCE: This proposal is critically concerned with the issue of public health in that its long term goal is to provide a drug delivery platform for providing sustained release of molecules to treat intraocular diseases. The short term goal is whether polymer nanofibers can deliver DNA to knock down expression of TGF-¿ in cell culture models of intraocular scarring disorders including corneal scarring, post-surgical scarring, and proliferative vitreoretinopathy.

描述（由申请人提供）：虽然在过去十年中已经出现了许多用于治疗眼部疾病的新治疗分子，但它们在临床中有效应用所面临的主要问题之一是药物递送通常具有较短的半衰期。在眼内给药的情况下，这通常意味着患者依从性差且成本高，因此，在数月至数年内控制释放治疗分子预计会对患者护理和治疗产生重大影响。眼部疾病的治疗。该提案的重点是研究用于眼内递送治疗分子的微米级和纳米级聚合物纤维。纤维比其他纳米级和微米级递送载体具有优势，因为如果出现并发症，它们可以移动或移除。 与多种眼部疾病相关的主要目标之一是转化生长因子-β (TGF-¿)，这些疾病包括角膜疤痕、白内障和青光眼手术后的疤痕，以及增殖性玻璃体视网膜病变。通过使用反义寡核苷酸、适体和其他小分子药物的表达及其效果，在针对 TGF-¿ 的 siRNA 治疗中取得了一些成功。虽然 siRNA 也已被证明是有效的，但由于其对普遍存在的 RNA 酶的敏感性以及需要大量分子才能在体内有效敲除，因此 siRNA 作为长期疗法受到限制。编码 siRNA 以敲除 TGF-¿ 的 DNA DNA 的表达解决了稳定性问题，因为 DNA 比 RNA 稳定得多，并且解决了药物量的问题，因为单个 DNA 分子可以转录成许多 siRNA。 本研究的目的是开发一种微型药物输送平台，该平台可以轻松植入以治疗多种眼部疾病，具体而言，本研究的假设是所提出的治疗性 DNA 可以从聚合物纳米纤维中掺入和释放。释放的 DNA 会导致眼细胞模型中的表达下调，该假设将在三个具体目标中进行测试：1) 评估静电纺丝形成的纳米纤维释放的 DNA 的生物活性，2) 在不稳定的 GFP 模型中评估。释放的 DNA 在敲低表达中的作用，以及 3) 在人角膜成纤维细胞模型中评估释放的 DNA 在敲低 TGF-¿选择这种眼部模型是因为研究人员对其很熟悉，但是，能够成功递送针对一种疾病的治疗性 DNA 的药物递送平台可以快速应用于其他疾病的治疗，例如湿型黄斑中的 VEGF 敲低。变性，或糖尿病性视网膜病变。 公共健康相关性：该提案非常关注公共健康问题，因为其长期目标是提供持续释放分子以治疗眼内疾病的药物输送平台，短期目标是聚合物纳米纤维是否可以输送DNA。抑制 TGF-¿ 的表达用于眼内疤痕疾病的细胞培养模型，包括角膜疤痕、术后疤痕和增殖性玻璃体视网膜病变。