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.

青光眼是治疗的顽固性，由此产生的视神经损伤是不可逆的。所有人口统计 容易受到青光眼的影响，尤其是老年人，非裔美国人，西班牙裔，糖尿病患者和家庭 青光眼史。约有300万美国人患有青光眼，美国政府失去了估计 每年30亿美元的税收收入，福利支出和支持这些补贴所需的医疗补贴 公民。非常需要对青光眼进行长期治疗，因为由于青光眼引起的视力丧失是 美国失明的第二大主要原因。当前可用的药理和外科手术 青光眼治疗具有显着的局限性和副作用，其中包括全身反应 药物，患者不合规，眼感染，手术装置故障以及眼睛损伤。基本的 开角青光眼（POAG）很常见，其特征是水性幽默的排水不良。 传统的流出路径。在这里，我们提出了一个新颖的病毒交付工程委员会 机械敏感通道（EMC）作为青光眼的TM受损中的压力调节器。使用 这项申请专利的技术，我们已经证明了EMC是：（i）在培养的TM细胞中功能，（ii） 在TM细胞中成功转导的体内转导； （iii）有效降低眼部小鼠模型中的IOP 没有（i）非靶向表达的高血压（由qPCR测量）； （ii）眼损伤（通过OCT评估）； （iii）血浆中的炎性细胞因子（由ELISA检测）；或（iv）免疫细胞反应和（v）生存力的丧失 长期表达EMC的靶向细胞。 EMC是细菌拉伸的工程版本 - 激活的通道，该通道直接在细胞的膜脂质双层中感受张力，并在响应中， 瞬时打开其大型非特异性孔，用于反式/脊柱抗性。在压力阈值下，EMC可以释放 通过SC清除TM细胞的流体。另外，细胞体积和大小的减少将 扩大细胞间空间，并促进幽默的旁细胞流动。 EMC充当理想的排水 TM电池的阀门，因为它是一个相对较小的同源频道，不需要任何相关的 蛋白质或能源组装和功能。我们已经确定了激活的最佳EMC变体 使用压力夹在生理上相关的压力下。拟议研究的目的是进一步 基于自我融合的AAV携带工程机械敏感EMC开发产品 用于POAG基因治疗的通道（SCEMC）。该阶段II SBIR项目的目的将完成 通过三个特定目标。 AIM 1：量化SCEMC的长期稳定性和SCEMC在野生型大鼠中的安全性； AIM 2：评估启用SCEMC的长期IOP降低POAG模型的功效和AIM 3：GLP 野生型NHP中注射体内注射SCEMC的毒性和生物分布的研究。一个发展 通过该提案的成功，安全有效的单剂量长期治疗对青光眼的治疗将有所改善 许多美国人的生活。