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（由应用程序提供）：虽然过去十年来，许多用于治疗眼部疾病的新治疗分子已获得，但其在诊所中有效应用面临的主要问题之一是药物递送。通常，生物活性分子的体内半衰期短，需要重复给药。在眼内给药的情况下，这通常意味着患者的依从性差和成本高。因此，预计在几个月至几年的情况下，受控的治疗分子释放将对患者护理和眼科疾病的治疗产生重大影响。该建议的重点是研究微观和纳米级聚合物纤维，用于治疗分子的眼内递送。纤维比其他纳米和显微镜输送向量具有优势，因为如果出现并发症，它们可以移动或去除。 在几种眼部疾病中实施的主要目标之一是改变生长因子-Beta（TGF - - ）。这些疾病包括角膜疤痕，白内障后疤痕和青光眼手术以及增殖的玻璃体肾上腺病。通过使用反义寡核苷酸，适体和其他小分子药物来降低TGF-表达及其作用，在治疗这些疾病方面有些成功。尽管也证明针对TGF- - 的siRNA的递送是有效的，但由于其对无处不在的RNass的敏感性，因此siRNA被限制为长期治疗，并且需要大量分子在体内有效敲低。在这项研究中，我们建议检查DNA的传递，该DNA代码为siRNA敲低TGF-€表达。 DNA的递送解决了稳定性问题，因为DNA比RNA稳定得多，并且解决了药物量问题，因为单个DNA分子可以转录为许多siRNA。 拟议的研究的目的是开发一个微观药物输送平台，该平台很容易植入以治疗多种眼部疾病。该研究的特定假设是，可以从聚合物纳米纤维中掺入并释放提出的治疗性DNA，并且释放的DNA可以导致眼细胞模型中的表达敲低。该假设将在三个具体目的中进行检验：1）评估由静电纺丝形成的纳米纤维释放的DNA的生物活性，2）在释放DNA在敲低表达中的稳定的GFP模型评估效应中，在敲低表达中的效果，3）在人的角膜成纤维细胞模型中评估DNA在TGF-ENSPERAMENT中释放的DNA的效应。由于研究人员对其的熟悉，因此选择了这种眼模型。但是，可以迅速将可以成功地针对一种疾病提供治疗性DNA的药物输送平台用于治疗其他疾病，例如湿型黄斑变性中的VEGF敲低或糖尿病性视网膜病。 公共卫生相关性：该提案与公共卫生问题非常关注，因为它的长期目标是提供一个药物输送平台，以提供持续释放分子以治疗眼内疾病。短期目标是聚合物纳米纤维是否可以输送DNA以击倒TGF- - 在包括角膜疤痕，外科手术后疤痕和增殖的玻璃体蛋白质疗法的细胞培养模型中的TGF- - 表达。