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.

在干法相关的黄斑变性（AMD）中的地理萎缩（GA）的特征是变性 感光体，是约1500万人的新视力丧失的主要原因。没有治愈 可以停止变性或恢复视力丧失的疗法。我们已经开发了可激活的环境光 多特征蛋白（MCO-II）允许刺激视网膜神经节细胞（RGC）进行视力康复。 但是，这种基因治疗向具有GA患者的临床翻译将需要针对性地递送Opsin- 将基因编码到萎缩区域，而不会扰动剩余的功能性视网膜。因此，我们有 开发了一种基于近红外激光的有效方法，用于靶向基因递送到视网膜中。在这个 纳米增强光学输送（NOD）方法，我们利用基于表面等离子体共振的场 功能化的金纳米棒（FGNR）的增强以瞬时打孔膜以提供 分子。在第一阶段，我们证明了MCO-II的靶向体内光学递送到退化的视网膜中 小鼠在多个波长下使用点头。此外，我们对连续波（CW）和 纳米脉冲激光基于MCO-II质粒的点头，并确定优化的激光参数 视网膜的有效转染。由于点头，未观察到可检测到的眼损伤。此外， 体内MCO-II质粒点点后，视网膜免疫染色没有明显的细胞死亡。 电生理研究表明，MCO-II敏化细胞可通过光激活，可以视觉 诱发皮质活动。该阶段II提案的总体目标是开发组合点头产品 用于以安全的方式使退化视网膜中的RGC光敏性RGC并刺激光敏的RGC 通过环境光恢复。为了实现这一目标，我们实现了以下目的：（1）量化长期 小鼠模型中缺乏光感受器的稳定性和安全性； （2）评估目标视网膜的功能 使用NOD辅助MCO-II在小鼠和大鼠模型中对RGC进行重新启动后的区域；和 （3）毒性，生物分布和效力的GLP研究，在非人类中点头MCO-II质粒 灵长类动物（NHP）。该协作提案汇集了光学交付方面的互补专业知识， 光遗传学，眼科，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，以应对视网膜的挑战 退化。 NHP的安全/功效研究将在CRO设施进行。完成后 第二阶段，我们设想：（i）临床研究的点头开发，（ii）IND应用于FDA， （iii）与风险投资和制药公司合作进行商业化。该提议的成功 将导致一种新的临床方法，用于通过常规玻璃体内注射FGNR治疗GA患者 和MCO-II。基于点头的不可渗透外源物质的靶向递送（小分子， 蛋白质和基因）将使药物，疫苗和基因疗法受益。