DNA nanoparticle formulations for optimal ocular gene delivery

用于最佳眼部基因传递的 DNA 纳米颗粒配方

• 批准号: 8545860
• 负责人: Muna I. Naash
• 金额: $ 42.41万
• 依托单位: UNIVERSITY OF OKLAHOMA HLTH SCIENCES CTR
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-09-30 至 2015-09-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8545860
• 关键词: Accounting; Affect; Animals; Automobile Driving; Biodistribution; Biologic Characteristic; Biomedical Engineering; Blindness; Brain; Bypass; Caliber; Cell Nucleus; Cell membrane; Cells; Characteristics; Chemicals; Chemistry; Clinic; Clinical; Coupled; DNA; Data; Development; Diabetic Retinopathy; Disease; Dose; Drug Formulations; Elements; Engineering; Enhancers; Enzyme-Linked Immunosorbent Assay; Epithelial; Evaluation; Excipients; Exhibits; Eye; Face; Fluorescence; Foundations; Future; GRB10 gene; Gene Delivery; Gene Expression; Gene Transfer; Generations; Genes; Goals; Human; Injection of therapeutic agent; Interphase Cell; Ions; Length; Longevity; Luciferases; Lung; Lysine; Matrix Attachment Regions; Mediating; Methodology; Methods; Mitotic; Modeling; Molecular; Mus; Nanotechnology; Papio; Parkinson Disease; Pathway interactions; Penetration; Peptides; Performance; Photoreceptors; Physics; Plasmids; Polyethylene Glycols; Polylysine; Primates; Procedures; Proteins; Research; Retina; Retinal; Retinal Cone; Retinal Degeneration; Retinal Diseases; Retinal Ganglion Cells; Retinal Pigments; Retinitis Pigmentosa; Retinol Binding Proteins; Rodent; Route; Safety; Shapes; Solvents; Structure of retinal pigment epithelium; System; Technology; Testing; Therapeutic; Time; Tissues; Toxic effect; Toxicology; Transfection; Treatment Efficacy; Vertebral column; Viral Vector; Visual Pathways; Vitelliform macular dystrophy; clinical application; cystic fibrosis patients; density; design; effective therapy; gene delivery system; gene therapy; improved; intravitreal injection; mRNA Expression; macula; macular dystrophy; minimally invasive; mouse model; nanoparticle; non-viral gene therapy; nonhuman primate; novel; particle; polycation; postnatal; preclinical study; prevent; programs; promoter; protein expression; ranpirnase; response; retinal rods; single molecule; therapeutic gene; trafficking; transgene expression; uptake; vector; vision science

DESCRIPTION (provided by applicant): The goal of this program is to advance current DNA nanoparticle (NP) delivery and expression technologies to develop safe and effective therapies for important ocular disorders affecting the photoreceptor (PR) and retinal pigment epithelial (RPE) cells. The program will merge experts with molecular bioengineering, vision science, physics, and chemistry to accelerate essential steps for the generation of effective ocular non-viral gene therapy. The DNA NPs consist of single molecules of DNA compacted with lysine-PEG polycations and have a minimum diameter of 8-11 nm. Their small size, coupled with a specific uptake mechanism that efficiently traffics the NPs to the nucleus and bypasses the lysosomal degradation system, likely accounts for the ability of the NPs to robustly transfect post-mitotic, differentiated cells. The counter-ion used at the time of formulation determines the NP shape and both rod-like and ellipsoidal NPs show robust transfection of ocular cells, including RPE, PRs and retinal ganglion cells (RGCs). Moreover, murine RDS NPs have demonstrated partial phenotypic correction in a retinitis pigmentosa mouse model of RDS haploinsufficiency. Having previously shown proof-of-principle for the effective use of these NPs in the eye, in this application we will take the necessary steps to optimize the particles for clinical ocular use. First, we will optimize the NP formulation (NP shape, size, and chemical composition) for PR- and RPE-specific gene transfer (Aim #1). Second, we will engineer clinically-appropriate DNA vectors (Aim #2) to generate persistent, high levels of transgene expression. Thirds, we will test the ability of these NPs to target the macula in a non-human primate model (baboon) (Aim #3) including an assessment of whether non-invasive intravitreal delivery of NPs can transfect macular/foveal cones. This is a novel and critical development for clinical application of this technology, as many retinal degenerations target the macula. Furthermore, it is a step that cannot be modeled by a mouse system due to the absence of a macula in the rodent retina. We will also conduct toxicology and DNA biodistribution studies in baboons, including a detailed evaluation of brain visual pathways. Building on the documented ability of the current NP formulation to penetrate deep retinal layers, we hypothesize that this efficiency can be improved by the NP formulation optimization program detailed in Aim #1, enabling robust foveal cone gene transfer. In summary, results from this application will facilitate preclinical trial evaluations of DNA NPs for ocular gene delivery.

描述（由申请人提供）：该项目的目标是推进当前的DNA纳米颗粒（NP）传递和表达技术，以开发安全有效的治疗影响光感受器（PR）和视网膜色素上皮（RPE）细胞的重要眼部疾病。该项目将整合分子生物工程、视觉科学、物理和化学方面的专家，以加速产生有效的眼部非病毒基因治疗的关键步骤。DNA NPs由DNA单分子与赖氨酸-聚乙二醇聚合组成，最小直径为8- 11nm。它们的小尺寸，加上特定的摄取机制，有效地将NPs运输到细胞核，绕过溶酶体降解系统，可能解释了NPs强大转染有丝分裂后分化细胞的能力。配制时使用的反离子决定了