Targeted Nano-enhanced Optical Delivery of opsin for dry-AMD therapy

用于干性 AMD 治疗的视蛋白靶向纳米增强光传递

• 批准号: 10011324
• 负责人: Samarendra Kumar Mohanty
• 金额: $ 77.99万
• 依托单位: NANOSCOPE TECHNOLOGIES, LLC
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-06-01 至 2023-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10011324
• 关键词: Address; Age related macular degeneration; Age-Years; Animals; Area; Atrophic; Behavior; Behavioral; Biodistribution; Biology; Bioluminescence; Biometry; Blindness; Capital; Cell Death; Cell membrane; Cells; Characteristics; Clinical; Clinical Pathology; Clinical Research; Comparative Study; DNA; DNA analysis; Disease; Dose; Dose-Limiting; Elderly; Electrophysiology (science); Endotoxins; FDA approved; Feedback; Fluorescence Microscopy; Gene Delivery; Genes; Goals; Gold; Head Movements; Histopathology; Image; Immunohistochemistry; Individual; Lasers; Light; Methods; Methylnitrosourea; Modeling; Molecular Biology; Mus; Mycoplasma; Neurosciences; Nonexudative age-related macular degeneration; Ophthalmology; Opsin; Optics; Organ; Patients; Persons; Pharmacotherapy; Phase; Photoreceptors; Photosensitization; Physiologic pulse; Plasmids; Production; Proteins; Rattus; Residual state; Retina; Retinal Degeneration; Retinal Ganglion Cells; Safety; Sampling; Signal Transduction; Small Business Innovation Research Grant; Structure; Surface Plasmon Resonance; Therapeutic; Therapeutic Effect; Toxic effect; Toxicology; Transfection; Transgenes; Vaccine Therapy; Viral; Vision; Visual Cortex; base; clinical translation; commercialization; efficacy study; gene therapy; geographic atrophy; immunogenicity; in vivo; instrument; instrumentation; intravitreal injection; mouse model; nano; nanomaterials; nanorod; nanosecond; nonhuman primate; optogenetics; patient population; photoreceptor degeneration; product development; retinal stimulation; small molecule; success; systemic toxicity; targeted delivery; therapeutic gene; vision rehabilitation

Geographic atrophies (GA) in dry-age related macular degeneration (AMD) is characterized by degeneration of photoreceptors, and is the leading cause of new vision loss in ~15 million persons. There is neither a cure that can stop the degeneration nor a therapy to restore vision loss. We have developed ambient-light activatable multi-characteristic opsin (MCO-II) to allow stimulation of retinal ganglion cells (RGCs) for vision rehabilitation. However, clinical translation of such gene therapy to patients with GA will require targeted delivery of opsin- encoding genes into the atrophic regions without perturbing remaining functional retina. Therefore, we have developed a near-infrared laser based efficient method for in-vivo targeted gene delivery into retina. In this Nano-enhanced Optical Delivery (NOD) method, we utilize surface plasmon resonance based field enhancement by functionalized gold nanorods (fGNRs) to transiently perforate cell membrane to deliver the molecules. In the Phase I, we demonstrated targeted in-vivo optical delivery of MCO-II to degenerated retina in mice using NOD at multiple wavelengths. Further, we made comparative study of continuous wave (cw) and nanosecond pulsed laser based NOD of MCO-II plasmids and determined optimized laser parameters for efficient transfection of retina. No detectable ocular damage was observed due to NOD. Further, the immunostaining of retina after in-vivo NOD of MCO-II plasmids showed no noticeable cell death. Electrophysiology studies demonstrate that MCO-II sensitized cells are activatable by light, allowing visually evoked cortical activities. The overall goal of this Phase-II proposal is to develop the combination NOD product for photosensitizing RGCs in the degenerated retina in a safe manner and stimulating photosensitized RGCs by ambient light for vision rehabilitation. Towards this goal we have following aims: (1) Quantify long-term stability and safety of NOD in mice model lacking photoreceptors; (2) Evaluate functioning of targeted retinal regions after re-photosensitization of RGCs using NOD assisted MCO-II delivery in mice and rat models; and (3) GLP study of toxicity, biodistribution and efficacy of NOD-delivered MCO-II plasmids in non-human primates (NHPs). This collaborative proposal brings together complementary expertise in optical delivery, optogenetics, ophthalmology, instrument, molecular biology, nanomaterials, retina biology and function, neuroscience/behavior, electrophysiology, biostatistics, and toxicology to address the challenge in retinal degeneration. The safety/efficacy study in NHPs will be performed at CRO facility. Upon completion of the Phase II we envision to advance: (i) NOD product development for clinical studies, (ii) IND application to FDA, and (iii) partnering with venture capital and Pharma company for commercialization. Success of this proposal will lead to a new clinical approach for treating patients with GA by conventional intravitreal injection of fGNRs and MCO-II. The NOD based targeted delivery of impermeable exogenous materials (small molecules, proteins and genes) will benefit drug, vaccine and gene therapy.

干性年龄相关性黄斑变性（AMD）中的地图状萎缩（GA）的特征在于 光感受器，并且是约1500万人新视力丧失的主要原因。没有治愈的方法， 既不能阻止退化，也没有恢复视力的治疗方法。我们开发了环境光激活 多特征视蛋白（MCO-II），以允许刺激视网膜神经节细胞（RGC）用于视力康复。 然而，这种基因疗法对GA患者的临床转化将需要靶向递送视蛋白- 编码基因进入萎缩区域，而不干扰剩余的功能性视网膜。所以我们 开发了一种基于近红外激光的有效方法，用于体内靶向基因递送到视网膜中。在这 纳米增强光传输（NOD）方法，我们利用表面等离子体共振为基础的领域 通过功能化的金纳米棒（fGNRs）增强瞬时包裹细胞膜以递送 分子。在第一阶段，我们证明了MCO-II的靶向体内光学传递到退化的视网膜， 小鼠使用NOD在多个波长。并对连续波（cw）和 基于纳秒脉冲激光的MCO-II质粒的NOD，并确定了优化的激光参数， 视网膜的高效转染。未观察到NOD导致的可检测的眼损伤。此夕h MCO-II质粒的体内NOD后视网膜的免疫染色显示没有明显的细胞死亡。 电生理学研究表明，MCO-II致敏细胞可被光激活，从而使视觉 诱发皮层活动。本第二阶段提案的总体目标是开发组合NOD产品 用于以安全的方式使变性视网膜中的RGC光敏化并刺激光敏化RGC 通过环境光进行视力康复。为了实现这一目标，我们有以下目标：（1）量化长期 NOD在缺乏光感受器小鼠模型中的稳定性和安全性;（2）评价靶向视网膜的功能 在小鼠和大鼠模型中使用NOD辅助的MCO-II递送对RGC进行再光敏化后的区域;以及 (3)NOD递送的MCO-II质粒在非人中的毒性、生物分布和功效的GLP研究 灵长类动物（NHP）。这项合作提案汇集了光学传输方面的互补专业知识， 光遗传学，眼科学，仪器，分子生物学，纳米材料，视网膜生物学和功能， 神经科学/行为，电生理学，生物统计学和毒理学，以解决视网膜病变的挑战 退化NHP的安全性/疗效研究将在CRO机构进行。完成后 第二阶段，我们设想推进：（i）NOD产品的临床研究开发，（ii）IND申请FDA， 及（iii）与风险投资及医药公司合作进行商业化。该提案的成功 将导致通过常规玻璃体内注射fGNRs治疗GA患者的新临床方法 MCO-II。基于NOD的非渗透性外源物质（小分子， 蛋白质和基因）将有利于药物、疫苗和基因治疗。