Evaluation of the lysosomal protease tripeptidyl peptidase 1 as a potential therapeutic for Alzheimer Disease

溶酶体蛋白酶三肽基肽酶 1 作为阿尔茨海默病潜在治疗剂的评估

• 批准号: 9808153
• 负责人: PETER LOBEL
• 金额: $ 19.88万
• 依托单位: RBHS-ROBERT WOOD JOHNSON MEDICAL SCHOOL
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-08-15 至 2021-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9808153
• 关键词: Abeta clearance; Acids; Affect; Age; Alzheimer' s Disease; Alzheimer' s disease model; Amyloid beta-Protein; Amyloid beta-Protein Precursor; Animals; Attenuated; Axon; Behavioral; Benchmarking; Biologic Development; Biological Assay; Biological Response Modifier Therapy; Blood; Blood - brain barrier anatomy; Blood Circulation; Brain; CASP1 gene; Caspase; Cathepsins B; Cerebrospinal Fluid; Child; Clinical; Cognitive deficits; Collaborations; Complement; Deposition; Development; Disease; Disease Progression; Drug Targeting; Enzyme-Linked Immunosorbent Assay; European; Evaluation; Excision; Exhibits; Female; Future; Genetic; Goals; Hippocampus (Brain); Human; Human Amyloid Precursor Protein; Immunohistochemistry; Impairment; Inherited; Investigation; Investments; J20 mouse; Jansky-Bielschowsky Disease; Laboratories; Laboratory Animals; Learning; Lymph; Lysosomal Storage Diseases; Lysosomes; Measures; Mediating; Medicine; Memory; Methods; Microglia; Modeling; Mus; Mutation; Nerve Degeneration; Outcome; Pathogenicity; Pathway interactions; Peptide Hydrolases; Peptides; Pharmaceutical Preparations; Phenotype; Play; Process; Production; Proteins; PubMed; Recombinant Proteins; Recombinants; Reporting; Role; Route; Severity of illness; Study models; System; Testing; Therapeutic; Transgenes; Transgenic Mice; V717F; abeta accumulation; age related; base; behavior measurement; behavioral study; brain cell; cell type; conditioned fear; disease phenotype; effective therapy; experience; extracellular; familial Alzheimer disease; improved; lysosomal proteins; male; medical schools; morris water maze; mouse model; nervous system disorder; neuron loss; novel; novel strategies; object recognition; overexpression; prevent; therapeutic protein; therapeutic target; trafficking; transgenic model of alzheimer disease; treatment group; tripeptidyl aminopeptidase

PROJECT SUMMARY/ABSTRACT Despite extensive efforts and investment, there is no cure for Alzheimer disease (AD) or effective treatment to slow progression of this devastating disorder. One potential therapeutic strategy is to promote degradation of amyloid beta (Aβ), whose accumulation in the brain is associated with and may be integral to the disease process. We have recently obtained strong evidence that the lysosomal protease tripeptidyl peptidase 1 (TPP1) plays an important role in degradation of Aβ fibrils. We hypothesize that increasing activity of TPP1 will promote degradation of Aβ, slowing or preventing its accumulation in the brain with subsequent therapeutic benefits for AD. We will test this hypothesis using two complementary approaches in the J20 mouse, a transgenic AD model that overproduces human Aβ and exhibits age-dependent plaque accumulation and cognitive deficits. First, in a genetic proof-of-principle study, we will cross the J20 mouse with our newly created transgenic mouse that constitutively overexpresses mouse TPP1 (~10-fold higher activity than endogenous levels). Second, we will use a peptide-mediated method to deliver recombinant human TPP1 protein from the bloodstream across the blood-brain barrier into the brain of the J20 mouse. Treatment groups and controls will each contain 30 animals with equal numbers of males and females. For both Aims, we will employ identical approaches to evaluate the effect of elevated TPP1 activity on the AD phenotype. At the age of seven months, we will analyze treated mice and controls using the Morris water maze, novel object recognition and fear conditioning assays. At eight months, the mice will be euthanized and levels of soluble and insoluble Aβ measured in brain extracts by ELISA and plaque measured in cortex and hippocampus by immunohistochemistry. If TPP1 augmentation has a positive effect on AD phenotype in the J20 mouse, this would provide a strong rationale to explore this further with the long-term goal of developing an effective therapy for AD. Additional studies in laboratory animals would be required prior to initiation of trials in humans. Looking forward, it is worth noting that targeting protein-based drugs across the blood-brain and/or cerebrospinal fluid-brain barrier continues to be a major obstacle for the development of biologic therapeutics for AD and other neurological disorders. However, in the case of TPP1, delivery of recombinant protein to the brain by intracerebroventricular administration has been approved by both the Federal Drug Administration and the European Medicines Agency for treatment of a neurodegenerative lysosomal storage disease. A similar delivery method should be feasible for AD and may allow repurposing of an existing drug.

项目摘要/摘要 尽管进行了广泛的努力和投资，但仍无法治愈阿尔茨海默氏病（AD）或有效治疗 这种毁灭性疾病的进展缓慢。一种潜在的理论策略是促进退化 淀粉样蛋白β（Aβ），其在大脑中的积累与疾病相关，并且可能是不可或缺的 过程。我们最近获得了有力的证据，表明溶酶体蛋白酶三肽肽1（TPP1） 在Aβ原纤维降解中起重要作用。我们假设TPP1的活动增加将促进 Aβ的降解，减慢或防止其在大脑中积累，并随后的治疗益处 广告。我们将使用J20小鼠（转基因AD模型）中的两种完整方法检验该假设 这过度产生了人Aβ，并表现出年龄依赖性的斑块积累和认知缺陷。 首先，在遗传学证明研究中，我们将与新创建的转基因小鼠越过J20小鼠 组成型过表达小鼠TPP1（比内源性水平高约10倍）。第二，我们 将使用肽介导的方法从血液中传递重组人TPP1蛋白 血脑屏障进入J20小鼠的大脑。治疗组和对照组将包含30个 雄性和女性数量相等的动物。对于这两个目标，我们将采用相同的方法来 评估TPP1活性升高对AD表型的影响。在七个月的时候，我们将分析 使用莫里斯水迷宫，新颖的对象识别和恐惧调节测定法对小鼠和对照进行了治疗。 在八个月时，小鼠将被安乐死，并在脑提取物中测得的固体和不溶性Aβ水平 通过免疫组织化学在皮质和海马中测量的ELISA和斑块。 如果TPP1增强对J20小鼠中的AD表型有积极影响，则可以提供强大的 基本原理以开发有效的AD疗法的长期目标，以进一步探讨这一点。额外的 在开始人类试验之前，需要进行实验动物的研究。展望未来，值得 注意到靶向基于蛋白质的药物在血脑和/或脑脊液 - 脑屏障中的靶向 仍然是开发AD和其他神经系统生物疗法的主要障碍 疾病。然而，在TPP1的情况下，脑室内脑室递送重组蛋白向大脑递送 政府已获得联邦药物管理局和欧洲药品局的批准 用于治疗神经退行性溶酶体储存疾病。类似的交货方法应该是可行的 对于AD，可能允许重新使用现有药物。