Topical delivery of nanoencapsulated plasmid DNA to posterior ocular targets

将纳米封装的质粒 DNA 局部递送至眼后部目标

• 批准号: 8252690
• 负责人: John M Nickerson
• 金额: $ 15.9万
• 依托单位: GENESEGUES, INC.
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-03-01 至 2014-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8252690
• 关键词: Adverse effects; Age related macular degeneration; Animals; Anterior eyeball segment structure; Apoptosis; Attenuated; Binding; Blindness; Carbohydrates; Cell Culture Techniques; Cell Nucleus; Cell Surface Receptors; Cells; Cessation of life; Clinic; Clinical Treatment; Clinical Trials; Complex; DNA; DNA delivery; Data; Dependovirus; Disease; Dose; Effectiveness; Electroporation; Eye; Eyedrops; Film; Gene Delivery; Gene Expression; Gene Therapy Agent; Genes; Glaucoma; Hereditary Disease; Hyaluronan; Immune response; Inflammation; Inflammatory Response; Injection of therapeutic agent; Laboratories; Life; Ligands; Longevity; Measures; Modeling; Molecular Weight; Morphology; Mus; Outcome Measure; Pathology; Patients; Permeability; Pharmaceutical Preparations; Phase; Plasmids; Prevention; Procedures; Property; Proteins; Protocols documentation; Public Health; RPE65 protein; Rattus; Regimen; Reporter; Reporter Genes; Research; Retina; Retinal Degeneration; Retinal Dystrophy; Safety; Shapes; Small Business Innovation Research Grant; Structure of retinal pigment epithelium; Technology; Testing; Therapeutic; Tissues; Topical application; Viral Vector; Vision; Work; absorption; capsule; design; dosage; effective therapy; expression vector; gene function; gene replacement; gene therapy; in vivo; inflammatory marker; mouse model; nanocapsule; nanoencapsulated; nanometer; novel; phase 1 study; phase 2 study; plasmid DNA; prevent; standard of care; subretinal injection; trafficking; vector

DESCRIPTION (provided by applicant): Diseases such as age related macular degeneration (AMD), other retinal dystrophies, and glaucoma cause a large proportion of blindness worldwide. A critical barrier in treating these diseases is the inability to efficaciously, economically, and safely deliver drugs and gene therapy DNA constructs to the retina and the retinal pigment epithelium (RPE). Topical delivery by eye drops has been ineffective because of permeability and absorption barriers of the anterior segment of the eye. Thus, current delivery approaches for these diseases entail costly injections of the eye using viral vectors that in some clinical trials have demonstrated dangerous side effects. The broad, long-term objective of this project is to develop a safe, economical, and effective gene delivery approach using novel, nonviral sub-50 nanometer (s50) capsule technology that is cell-specific and traffics to the nucleus without endosomal entrapment. Our preliminary data indicate that these nanocapsules administered as topical eye drops onto the tear film of living rats result in delivery and expression of gene vectors in cells of the retina and RPE. Topical application resulting in DNA delivery to posterior ocular targets is surprising and potentially transformative. If brought to the clinic, it should provide safe and economical treatment for many blinding diseases. In this Phase 1 study, we propose to optimize the topical s50-capsule dosing regimen in terms of safety and efficacy using easily-assessed fluorescent plasmid DNA expression vectors. We then will compare expression levels and inflammatory markers of the optimized topical s50-capsule dosing regimen, with an ocular injection protocol that we have previously optimized. We will use the LCA2 mouse model of retinal degeneration. LCA2 mouse models are useful because they have severe vision deficits due to simple lack of a single gene product (RPE65), yet they have delayed morphological degeneration. Thus, successful delivery of replacement gene vector results in creation of visual function and prevention of degeneration. If the results of this Phase I study are encouraging, we will propose Phase II studies using the novel s50 capsule technology to deliver therapeutic RPE65 expression vectors to mice deficient in RPE65, and assess target gene expression and function, visual function, and morphology. PUBLIC HEALTH RELEVANCE: The relevance of this research to public health is potentially very high. A large proportion of blindness is caused by retina and retinal pigment epithelium (RPE) pathologies. Several therapeutic DNA constructs and drugs are effective treatments for these diseases in animal and cell culture models, but are of limited use in the clinic because they cannot be delivered to target tissue and cells efficiently and safely. The aim of this project is to develop a safe and effective way to deliver these treatments.

描述(申请人提供)：老年性黄斑变性(AMD)、其他视网膜营养不良和青光眼等疾病在全球范围内导致很大比例的失明。治疗这些疾病的一个关键障碍是无法有效、经济和安全地将药物和基因治疗DNA结构输送到视网膜和视网膜色素上皮(RPE)。由于眼球前段的渗透性和吸收障碍，滴眼液局部给药无效。因此，目前治疗这些疾病的方法需要使用病毒载体进行昂贵的眼部注射，而在一些临床试验中，病毒载体已显示出危险的副作用。该项目广泛的、长期的目标是开发一种安全、经济和有效的基因传递方法，使用新型的非病毒亚50纳米(S50)胶囊技术，这种技术是细胞特异性的，可以在没有内体包裹的情况下输送到细胞核。我们的初步数据表明，这些纳米胶囊作为局部滴眼液滴到活着的大鼠泪膜上，导致基因载体在视网膜和RPE细胞中的传递和表达。局部应用导致将DNA输送到眼后靶点是令人惊讶的，并具有潜在的变革性。如果推向临床，它将为许多致盲疾病提供安全、经济的治疗。在这项第一阶段的研究中，我们建议使用易于评估的荧光质粒DNA表达载体在安全性和有效性方面优化S50胶囊的局部给药方案。然后，我们将比较优化的S50胶囊局部给药方案和我们之前优化的眼部注射方案的表达水平和炎症标志物。我们将使用LCA2小鼠的视网膜变性模型。LCA2小鼠模型是有用的，因为它们由于简单地缺乏单一基因产物(RPE65)而具有严重的视力障碍，但它们延缓了形态退化。因此，成功地传递替换基因载体会导致视觉功能的产生和防止退化。如果这项第一阶段研究的结果令人鼓舞，我们将提出第二阶段研究，使用新型S50胶囊技术将治疗性RPE65表达载体运送到RPE65缺陷小鼠，并评估目标基因的表达和功能、视觉功能和形态。 公共卫生相关性：这项研究与公共卫生的相关性可能非常高。很大一部分失明是由视网膜和视网膜色素上皮(RPE)病理引起的。在动物和细胞培养模型中，几种治疗性DNA构建物和药物是治疗这些疾病的有效方法，但由于它们不能有效和安全地输送到靶组织和细胞，因此在临床上的应用有限。该项目的目的是开发一种安全有效的方法来提供这些治疗。