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

基本信息

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
项目类别：
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

项目摘要

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) 中的地理萎缩 (GA) 的特征是黄斑变性光感受器，是约 1500 万人新视力丧失的主要原因。没有治愈方法不能阻止退化，也不能治疗恢复视力丧失。我们开发了环境光可激活多特征视蛋白（MCO-II）可刺激视网膜神经节细胞（RGC）以实现视力康复。然而，将此类基因疗法临床转化为 GA 患者需要靶向递送视蛋白将基因编码到萎缩区域而不干扰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，用于将体内靶向基因递送至视网膜。在这个纳米增强光传输（NOD）方法，我们利用基于表面等离子体共振的场通过功能化金纳米棒（fGNR）增强，瞬时穿孔细胞膜以传递分子。在第一阶段，我们展示了 MCO-II 定向体内光学递送至退化视网膜小鼠在多个波长下使用 NOD。此外，我们对连续波（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有效转染。没有观察到由 NOD 引起的可检测到的眼部损伤。此外， MCO-II 质粒体内 NOD 后的视网膜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，从而使视觉诱发皮质活动。该二期提案的总体目标是开发组合NOD产品以安全的方式刺激退化视网膜中的光敏 RGC 并刺激光敏 RGC 通过环境光进行视力康复。为实现这一目标，我们有以下目标：（1）量化长期目标 NOD在缺乏光感受器的小鼠模型中的稳定性和安全性； (2) 评估目标视网膜的功能在小鼠和大鼠模型中使用 NOD 辅助 MCO-II 递送对 RGC 进行重新光敏化后的区域；和 (3) NOD 递送的 MCO-II 质粒在非人类体内的毒性、生物分布和功效的 GLP 研究灵长类动物（NHP）。该合作提案汇集了光学传输领域的互补专业知识，光遗传学、眼科、仪器、分子生物学、纳米材料、视网膜生物学与功能、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 的安全性/有效性研究将在 CRO 设施中进行。完成后我们设想推进第二阶段：(i) 用于临床研究的 NOD 产品开发，(ii) 向 FDA 申请 IND， (iii) 与风险投资和制药公司合作进行商业化。本提案的成功将通过传统玻璃体内注射 fGNR 为治疗 GA 患者带来一种新的临床方法和MCO-II。基于 NOD 的不可渗透外源材料（小分子、蛋白质和基因）将有利于药物、疫苗和基因治疗。