Mechanosensitive Channel based Pressure-Modulating Gene Therapy for Glaucoma Treatment

基于机械敏感通道的压力调节基因疗法治疗青光眼

• 批准号: 10384600
• 负责人: ADNAN Ismail DIBAS
• 金额: $ 71.69万
• 依托单位: NANOSCOPE TECHNOLOGIES, LLC
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-30 至 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10384600
• 关键词: Address; African American population; Aftercare; Age; American; Americas; Anti-Inflammatory Agents; Antibodies; Aqueous Humor; Biodistribution; Biology; Biomedical Engineering; Biometry; Blindness; Capsid; Cell Size; Cell Volumes; Cells; Clinical; Clinical Pathology; Closure by clamp; DNA; DNA analysis; Development; Disease; Dose; Dose-Limiting; Drainage procedure; Elderly; Electrophysiology (science); Energy-Generating Resources; Engineering; Enzyme-Linked Immunosorbent Assay; Equipment Malfunction; Extracellular Space; Eye; Eye Infections; FDA approved; Family; Genes; Glaucoma; Goals; Government; Guidelines; Healthcare; Hispanic Populations; Histopathology; Homo; Immune; Immunohistochemistry; Impairment; Inflammatory; Injections; Legal patent; Life; Lipid Bilayers; Liquid substance; Measurement; Measures; Membrane; Molecular; Mus; Ocular Hypertension; Operative Surgical Procedures; Ophthalmology; Optic Nerve; Organ; Outcome; Pathway interactions; Patient Noncompliance; Peripheral Blood Mononuclear Cell; Persons; Pharmaceutical Preparations; Pharmacology; Phase; Physiologic Intraocular Pressure; Physiological; Plasma; Primary Open Angle Glaucoma; Production; Proteins; Quality of life; Rattus; Reaction; Recording of previous events; Research; Residual state; Retina; Rodent Model; Safety; Sampling; Serum; Signal Transduction; Small Business Innovation Research Grant; Stretching; Suction; Taxes; Technology; Therapeutic; Therapeutic Effect; Toxic effect; Transgenes; Transgenic Organisms; Variant; Viral; Visual Cortex; aqueous humor flow; base; cytokine; demographics; diabetic; gene therapy; improved; in vivo; mechanical force; mouse model; nerve damage; novel; pressure; response; sensor; sex; side effect; success; systemic toxicity; ultrasound; vector

Glaucoma is recalcitrant to treatment and the resulting optic nerve damage is irreversible. All demographics are susceptible to glaucoma, especially the elderly, African-Americans, Hispanics, diabetics and families with a history of glaucoma. About three million Americans have glaucoma and the US government loses an estimated 3 billion USD each year in lost tax revenue, benefit payouts and healthcare subsidies needed to support these citizens. There is great need for a long-lasting treatment for glaucoma, because vision loss due to glaucoma is the second leading cause of blindness in America. The currently available pharmacological and surgical treatments for glaucoma have significant limitations and side-effects, which include, systemic reactions to medications, patient non-compliance, eye infections, surgical device failure, and damage to the eye. Primary Open Angle Glaucoma (POAG) is common and is characterized by poor drainage of aqueous humor through the conventional outflow pathway. Here, we put forth a novel commission for virally delivered Engineered Mechanosensitive Channel (EMC) as a pressure modulator in the impaired TM of glaucomatous eyes. Using this patent pending technology, we have demonstrated that EMC is: (i) functional in cultured TM cells, (ii) successfully transduced in vivo in TM cells; and (iii) effective in lowering the IOP in a mouse model of ocular hypertension without (i) non-targeted expression (measured by QPCR); (ii) ocular damage (assessed by OCT); (iii) inflammatory cytokines in plasma (detected by ELISA); or (iv) immune cell response and (v) loss of viability of targeted cells expressing EMC over long period. EMC is an engineered version of the bacterial stretch- activated channel, which directly senses tension in the membrane lipid bilayer of cells and in response, transiently opens its large non-specific pore for trans/paracytosis. At its pressure threshold, EMC can release fluid from TM cells to be cleared through SC. Additionally, the resulting decrease in cell volume and size will widen intercellular spaces and facilitate the paracellular flow of aqueous humor. EMC acts as an ideal drainage valve for TM cells, because it is a relatively small homo-oligomeric channel and does not need any associated proteins or energy sources to assemble and function. We have identified optimal EMC variant that is activated at physiologically relevant pressures using pressure clamp. The goal of the proposed research is to further develop the product based on the self-complementary AAV carried engineered mechanosensitive EMC channel (scEMC) for gene therapy of POAG. The objective of this phase-II SBIR project will be accomplished through three specific aims. Aim 1: Quantify long-term stability of scEMC and safety of scEMC in wild type rats; Aim 2: Evaluate scEMC-enabled long-term IOP lowering Efficacy in rodent models of POAG and Aim 3: GLP Study of toxicity and Biodistribution of intracamerally-injected scEMC in wild-type NHPs. The development of a safe effective single-dose long-lasting treatment for glaucoma via the success of this proposal would improve the lives of many Americans.

青光眼是顽固性的治疗，由此导致的视神经损害是不可逆转的。所有人口统计数据 易患青光眼，尤其是老年人、非裔美国人、西班牙裔、糖尿病患者和患有青光眼的家庭 青光眼病史。约300万美国人患有青光眼，美国政府估计 每年损失30亿美元的税收、福利支出和医疗补贴 公民们。青光眼的长期治疗非常需要，因为青光眼导致的视力丧失是 在美国，失明的第二大原因。目前可用的药理学和外科手术 青光眼的治疗有很大的局限性和副作用，包括全身反应 药物、患者不依从、眼睛感染、手术器械故障和眼睛损伤。主要 开角型青光眼(POAG)是一种常见的青光眼，其特点是房水排出不良。 传统的流出路径。在这里，我们提出了一个新的委托病毒交付工程 机械敏感通道(EMC)在青光眼TM受损中的压力调节作用vbl.使用 这项正在申请专利的技术，我们已经证明了EMC是：(I)在培养的TM细胞中具有功能，(Ii) 在TM细胞中成功地体内转导；以及(Iii)在小鼠眼部模型中有效地降低眼压 高血压无(I)非靶向表达(用定量聚合酶链式反应测定)；(Ii)眼损伤(用OCT评估)； (Iii)血浆中的炎性细胞因子(用酶联免疫吸附试验检测)；或(Iv)免疫细胞反应和(V)丧失活力 长时间表达EMC的靶细胞的数量。EMC是细菌拉伸的工程化版本- 激活的通道，它直接感受细胞膜脂双层的张力并做出反应， 瞬间打开其大的非特异性毛孔，用于反式/旁向细胞凋亡。在压力临界点，EMC可以释放 TM细胞中的液体要通过SC进行清理。此外，由此导致的细胞体积和大小的减少将 拓宽细胞间隙，促进房水的细胞旁流动。EMC充当理想的引流设备 TM细胞的瓣膜，因为它是一个相对较小的同源低聚通道，不需要任何相关的 蛋白质或能量来源来组装和发挥作用。我们已经确定了激活的最佳EMC变体 在生理相关压力下使用压力钳。拟议研究的目标是进一步 基于自补型AAV搭载工程化机械敏感EMC的产品开发 通道(ScEMC)用于POAG的基因治疗。二期SBIR项目的目标将会实现 通过三个具体目标。目的1：定量scEMC在野生型大鼠体内的长期稳定性和安全性； 目的2：在POAG和AIM 3：GLP啮齿动物模型上评价scEMC启用的长期降低眼压的效果 SCEMC在野生型NHP内注射的毒性及生物分布研究开发一种新的 通过这项提议的成功，安全有效的单剂长效青光眼治疗将会改善 许多美国人的生活。