Activation of Soluble Adenylyl Cyclase as a Novel Therapeutic Strategy for the Treatment of Age-Related Neurodegenerative Disorders

激活可溶性腺苷酸环化酶作为治疗年龄相关神经退行性疾病的新治疗策略

• 批准号: 10443843
• 负责人: Thomas Rossetti
• 金额: $ 3.74万
• 依托单位: WEILL MEDICAL COLL OF CORNELL UNIV
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-08-11 至 2023-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10443843
• 关键词: Acids; Adenosine Triphosphate; Adenylate Cyclase; Affect; Alzheimer' s Disease; Alzheimer' s disease model; Alzheimer' s disease pathology; Alzheimer' s disease patient; Amyloid; Autophagocytosis; Biological; Biological Assay; Biological Sciences; Biology; Cell Aging; Cell model; Cells; Chemicals; Cyclic AMP; Cyclization; Defect; Degradation Pathway; Disease; Disease Progression; Environment; Enzyme-Linked Immunosorbent Assay; Exhibits; FDA approved; Fibroblasts; Functional disorder; Genetic; Hydrolase; Immune; In Vitro; Individual; Knock-out; Label; Length; Libraries; Life Expectancy; LysoTracker; Lysosomes; Mammalian Cell; Measures; Methods; Microglia; Modeling; Neurodegenerative Disorders; Organelles; Parkinson Disease; Pathogenesis; Pathologic; Patients; Pharmacology; Phenotype; Physiological; Physiological Processes; Play; Population; Protein Isoforms; Public Health; Research; Role; Second Messenger Systems; Signal Transduction; Signal Transduction Pathway; Specificity; Testing; Western Blotting; age related; age related neurodegeneration; aging population; alkalinity; base; brain cell; cell age; cell type; cellular engineering; effective therapy; experimental study; familial Alzheimer disease; high throughput screening; human old age (65+); in vitro Assay; inhibitor; misfolded protein; mutant; novel; novel strategies; novel therapeutic intervention; overexpression; presenilin-1; protein aggregation; protein degradation; small molecule; therapeutically effective; tool; treatment strategy; Acids; Adenosine Triphosphate; Adenylate Cyclase; Affect; Alzheimer' s Disease; Alzheimer' s disease model; Alzheimer' s disease pathology; Alzheimer' s disease patient; Amyloid; Autophagocytosis; Biological; Biological Assay; Biological Sciences; Biology; Cell Aging; Cell model; Cells; Chemicals; Cyclic AMP; Cyclization; Defect; Degradation Pathway; Disease; Disease Progression; Environment; Enzyme-Linked Immunosorbent Assay; Exhibits; FDA approved; Fibroblasts; Functional disorder; Genetic; Hydrolase; Immune; In Vitro; Individual; Knock-out; Label; Length; Libraries; Life Expectancy; LysoTracker; Lysosomes; Mammalian Cell; Measures; Methods; Microglia; Modeling; Neurodegenerative Disorders; Organelles; Parkinson Disease; Pathogenesis; Pathologic; Patients; Pharmacology; Phenotype; Physiological; Physiological Processes; Play; Population; Protein Isoforms; Public Health; Research; Role; Second Messenger Systems; Signal Transduction; Signal Transduction Pathway; Specificity; Testing; Western Blotting; age related; age related neurodegeneration; aging population; alkalinity; base; brain cell; cell age; cell type; cellular engineering; effective therapy; experimental study; familial Alzheimer disease; high throughput screening; human old age (65+); in vitro Assay; inhibitor; misfolded protein; mutant; novel; novel strategies; novel therapeutic intervention; overexpression; presenilin-1; protein aggregation; protein degradation; small molecule; therapeutically effective; tool; treatment strategy

Project Summary/Abstract The worldwide population of individuals over the age of 65 is continuing to grow and is expected to double over the next 30 years. Due to this increase, an associated rise in age-related neurodegenerative diseases (NDs), such as Alzheimer’s (AD) and Parkinson’s disease, has been observed. Despite decades of research, there are currently no FDA-approved therapies that can stop or reverse disease progression. The accumulation of misfolded protein aggregates is a common feature of age-related NDs and is thought to be heavily involved in the pathophysiology of the diseases. Proteins aggregates can be cleared from the cell through autophagy, one of the major biological degradation pathways. Protein aggregates are degraded in the final step of autophagy, where they are delivered to cellular organelles known as lysosomes. Lysosomes maintain an acidic pH between 4-5 to maintain an optimal environment for acid hydrolases. In aging cells and in a cellular model of AD, lysosomes become less acidic, their hydrolases become less active and, as a result, there is a decrease in degradation through autophagy (i.e., autophagic flux) and an accumulation of undigested materials. In models of AD pathology, re-acidification of lysosomal pH, via addition of exogenous cAMP, reversed this phenotype. In line with these findings, we have shown that cAMP generated from a cytosolic adenylyl cyclase isoform, known as Soluble Adenylyl Cyclase (sAC), promotes lysosomal acidification. In addition, cells that do not express sAC show decreased autophagic flux. This, and other, physiological roles of sAC have been determined via the use of inhibitors and various genetic tools that were developed to study sAC biology. However, the “toolbox” that is currently used to study sAC is lacking a key component: a pharmacological activator of sAC. To identify a sAC activator, we conducted a high-throughput screen of 400,000 chemically diverse compounds. From this library we discovered 13 presumptive sAC activators. In this proposal, I describe the in vitro and cell-based assays that I will use to confirm, characterize, and further develop these 13 compounds as “first-of-their kind” small molecule activators selective for sAC. Using the newly discovered sAC activators, I will test the hypothesis that stimulating sAC can enhance lysosomal acidification and stimulate autophagy, and as a result, decrease accumulation of protein aggregates. If successful, these studies will validate small molecule sAC activators as a potential novel therapeutic strategy to treat neurodegenerative disorders.

项目摘要/摘要 65岁以上的全球个人人口正在继续增长，预计将两倍 在接下来的30年中。由于这种增加，与年龄有关的神经退行性疾病的增加 已经观察到（NDS），例如阿尔茨海默氏症（AD）和帕金森氏病。尽管进行了数十年的研究， 目前没有FDA批准的疗法可以阻止或逆转疾病的进展。积累 错误折叠的蛋白质聚集体是与年龄相关的NDS的共同特征，被认为涉及大量 在疾病的病理生理中。蛋白质聚集体可以通过自噬从细胞清除， 主要的生物降解途径之一。蛋白质聚集体在最后一步 自噬，将它们传递到称为溶酶体的细胞细胞器中。溶酶体保持酸性 pH值在4-5之间，以维持酸水解酶的最佳环境。在衰老细胞和在细胞模型中 AD，溶酶体变得较少酸性，其水溶液的活性降低，因此，降低了 通过自噬（即自噬通量）和无灾的材料的积累来降解。在模型中 AD病理学，通过添加外源营地的溶酶体pH的重新二化，逆转了这种表型。在 与这些发现相符，我们表明cAMP是从胞质腺苷酸环化酶同工型产生的，已知 作为可溶性腺苷酸环化酶（SAC），促进溶酶体酸化。另外，不表达囊的细胞 显示自噬的减少。 SAC的此和其他物理角色已通过使用确定 开发用于研究囊生物学的抑制剂和各种遗传工具。但是，“工具箱”就是 目前用于研究SAC的目前缺乏关键组成部分：SAC的药物激活剂。识别囊 激活剂，我们进行了400,000种化学化合物的高通量屏幕。来自这个库 我们发现了13个推定的SAC激活剂。在此提案中，我描述了基于细胞和细胞的测定 我将用来确认，表征和进一步开发这13种化合物作为“首先类型”的小分子 激活剂选择SAC。使用新发现的SAC激活剂，我将测试刺激的假设 SAC可以增强溶酶体酸化并刺激自噬，因此，降低了 蛋白质聚集体。如果成功，这些研究将验证小分子囊活化剂作为潜在的新颖 治疗神经退行性疾病的治疗策略。