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

基本信息

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
项目类别：
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&apos s Disease Alzheimer&apos 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

项目摘要

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.

项目概要/摘要尽管付出了大量努力和投资，但阿尔茨海默病 (AD) 仍无法治愈，也无法有效治疗一种潜在的治疗策略是促进这种破坏性疾病的缓慢进展。 β 淀粉样蛋白 (Aβ)，其在大脑中的积累与该疾病有关，并且可能是该疾病不可或缺的一部分我们最近获得了强有力的证据表明溶酶体蛋白酶三肽基肽酶 1 (TPP1) 我们发现TPP1活性的增加会促进Aβ原纤维的降解。 Aβ 的降解，减缓或阻止其在大脑中的积累，从而产生后续的治疗效果我们将在 J20 小鼠（转基因 AD 模型）中使用两种互补的方法来检验这一假设。过量产生人类 Aβ，并表现出年龄依赖性斑块积累和认知缺陷。首先，在基因原理验证研究中，我们将 J20 小鼠与我们新创建的转基因小鼠杂交其次，我们持续过度表达小鼠 TPP1（活性比内源水平高约 10 倍）。将使用肽介导的方法将重组人 TPP1 蛋白从血液中输送到整个组织治疗组和对照组各包含 30 个进入 J20 小鼠大脑的血脑屏障。对于这两个目标，我们将采用相同的方法来实现雄性和雌性数量相等的动物。评估 TPP1 活性升高对 AD 表型的影响在七个月大时，我们将进行分析。小鼠和对照组使用莫里斯水迷宫、新物体识别和恐惧调节试验进行治疗。八个月时，小鼠将被安乐死，并测量大脑提取物中可溶性和不溶性 Aβ 的水平通过 ELISA 测定，并通过免疫组织化学测定皮质和海马中的斑块。如果 TPP1 增强对 J20 小鼠的 AD 表型有积极影响，这将提供强大的进一步探索这一点的理由是为了开发一种有效的 AD 疗法的长期目标。在开始人体试验之前需要对实验动物进行研究，展望未来，这是值得的。注意到靶向蛋白质药物穿过血脑和/或脑脊液脑屏障仍然是 AD 和其他神经系统疾病生物疗法开发的主要障碍然而，在 TPP1 的情况下，重组蛋白通过脑室内递送至大脑。管理已获得联邦药物管理局和欧洲药品管理局的批准用于治疗神经退行性溶酶体贮积症的类似递送方法应该是可行的。用于 AD 并可能允许重新利用现有药物。