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: 10292932
• 负责人: Thomas Rossetti
• 金额: $ 4.6万
• 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 2023-08-10
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10292932
• 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; Immunoblotting; 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; 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/antagonist; 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.

项目总结/摘要 全世界65岁以上的人口正在继续增长，预计将翻一番。 在接下来的30年里。由于这种增加，与年龄相关的神经退行性疾病的相关增加 (NDs)如阿尔茨海默氏病（AD）和帕金森氏病。尽管经过几十年的研究， 目前还没有FDA批准的可以阻止或逆转疾病进展的疗法。积累 错误折叠的蛋白质聚集体是年龄相关性ND的共同特征， 疾病的病理生理学。蛋白质聚集体可以通过自噬从细胞中清除， 主要的生物降解途径之一。蛋白质聚集体在最后一步降解， 自噬，在那里它们被递送到称为溶酶体的细胞器。溶酶体保持酸性 pH值在4-5之间，以保持酸性水解酶的最佳环境。在衰老细胞和衰老细胞模型中， AD时，溶酶体的酸性降低，其水解酶活性降低，结果， 通过自噬降解（即，自噬流）和未消化物质的积累。模型中 在AD病理学中，通过添加外源性cAMP使溶酶体pH重新酸化，逆转了这种表型。在 与这些发现一致，我们已经证明cAMP产生于胞浆腺苷酸环化酶同种型，已知 作为可溶性腺苷酸环化酶（sAC），促进溶酶体酸化。此外，不表达sAC的细胞 显示出自噬流减少。sAC的这种和其他生理作用已经通过使用 抑制剂和各种遗传工具，这些工具是为了研究sAC生物学而开发的。然而，“工具箱”， 目前用于研究sAC的药物缺乏一种关键成分：sAC的药理学激活剂。识别sAC 活化剂，我们对40万种化学上不同的化合物进行了高通量筛选。从库中 我们发现了13种可能的sAC激活剂在这个提议中，我描述了体外和基于细胞的测定， 我将用它来确认、表征和进一步开发这13种化合物作为“首个同类”小分子 对sAC具有选择性的活化剂。使用新发现的sAC激活剂，我将测试刺激 sAC可以增强溶酶体酸化并刺激自噬，从而减少 蛋白质聚集体。如果成功，这些研究将验证小分子sAC激活剂作为一种潜在的新型药物。 治疗神经退行性疾病的治疗策略。