Controlled expression of a cystatin C-peptide as therapy for Alzheimer's disease

半胱氨酸蛋白酶抑制剂 C 肽的受控表达作为阿尔茨海默病的治疗方法

• 批准号: 8323235
• 负责人: PHYLLIS Ann SHAW
• 金额: $ 18.01万
• 依托单位: ICAHN SCHOOL OF MEDICINE AT MOUNT SINAI
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-09-01 至 2014-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8323235
• 关键词: 1 year old; Affect; Alzheimer' s Disease; Alzheimer' s disease model; Amyloid; Area; Binding; Brain; C-Peptide; Cathepsins B; Chronic; Cleaved cell; Clinical Trials; Complex; Cysteine Protease; Data; Degenerative Disorder; Deposition; Development; Disease; Disease Progression; Dose; Encapsulated; Enzymes; Etiology; Family; Fluorescein-5-isothiocyanate; Fluorescence Microscopy; Functional disorder; Future; Gene Expression; Gene Mutation; Hemagglutinin; Hippocampus (Brain); Hormonal; Hormones; Human; Immunohistochemistry; Impaired cognition; Impairment; In Vitro; Incubated; Individual; Intervention; Learning; Life; Ligands; Link; Long-Term Potentiation; Mediating; Memory; Memory Loss; Methods; Mind; Mus; Mutation; Neocortex; Nerve Degeneration; Neurodegenerative Disorders; Neurofibrillary Tangles; Onset of illness; Pathology; Peptide Hydrolases; Peptides; Performance; Plasmids; Prevalence; Prevention; Preventive; Protein Precursors; Regulation; Relative (related person); Role; Senile Plaques; Slice; Synapses; Testing; Therapeutic; Transgenic Mice; Translating; Western Blotting; beta-site APP cleaving enzyme 1; cerebrovascular; citrate carrier; cognitive function; design; disease phenotype; disease-causing mutation; extracellular; familial Alzheimer disease; improved; in vivo; inhibitor/antagonist; morris water maze; mouse model; neuron loss; neurotoxicity; novel; particle; post gamma-globulins; presenilin-1; prevent; promoter; research study; response; secretase; treatment effect

DESCRIPTION (provided by applicant): Alzheimer's disease (AD), a fatal neurodegenerative disorder, afflicts close to 26.6 million people worldwide. It is characterized by accumulation in the brain of insoluble, extracellular amyloid plaques and intracellular neurofibrillary tangles considered to be, until recently, the main cause of brain dysfunction. This view has been recently reformulated to focus on soluble oligomeric Ab (oA?) aggregates, with emphasis on the role of A?42 oligomers, which are believed to cause synaptic dysfunction and eventual neuronal loss in the vulnerable regions of AD brains. Cathepsin B, a cysteine protease present in the brain, can degrade A? peptides, however, blocking its enzymatic activity in vivo with its specific inhibitor, cystatin C (CysC), rather than increasing, lowered A? peptide brain deposition in AD models. Because a portion of the CysC protein (aa101- 117), distinct from its enzyme inhibitory sequence, can bind to A? and inhibit fibril formation in vitro, we propose that generating sufficiently high levels of this peptide in vivo, might reduce A? oligomerization and neurotoxicity. We showed a) that synthetic CysC-A?-Binding Peptide (CysC-A?BP) binds specifically and preferentially to purified A?42, b) that in b AD mouse models, one with amyloid plaques and oA? (TgCRND8) and one with oA? only (APPE693Q), it co-localizes with the A? deposits in neocortex and hippocampus when incubated with brain sections in vitro, c) that similar co-localization is found 4 days after a single intranasal delivery of the peptide and d) that intranasal delivery of a CysS-GFP-expressing construct under a promoter that we have shown to be highly upregulated in the brain by several hormones produces detectable expression levels. We propose: 1. To identify conditions that generate optimal levels of CysC-A?BP in the brain. This will be done by comparing intranasal to intrathecal delivery of purified peptide and peptide-expressing construct, uncoated or encapsulated in microparticles, and to test which positive hormonal regulators of the CysS promoter optimally transactivate the CysC-A?BP construct. 2. To test whether high levels of CysC-A?BP will reduce A?-oligomer deposits in established AD and whether chronic treatment of mice before AD onset will prevent its development. Experimental conditions deemed most effective in A? oligomer reduction in response to CysC-A?BP will be utilized to test the effect this treatment has on improvement of learning and memory. By proposing to use an A?BP-fibril blocking CysC peptide, rather than the complete CysC protein, with its detrimental enzyme inhibitory activity, and by finding novel ways to deliver regulate and maximize CysC-A?BP expression in the brain, we expect to achieve beneficial effects that might be translated into therapeutic and preventive approaches to AD in humans.

描述（由申请人提供）：阿尔茨海默病（AD）是一种致命的神经退行性疾病，全世界有近2660万人患有此病。它的特点是在大脑中积累不溶性的细胞外淀粉样斑块和细胞内神经原纤维缠结，直到最近才被认为是脑功能障碍的主要原因。这一观点最近被重新表述为关注可溶性低聚Ab （oA?）聚集体，并强调A?的作用。42个低聚物，被认为在阿尔茨海默病大脑的脆弱区域导致突触功能障碍和最终的神经元丢失。组织蛋白酶B，一种存在于大脑中的半胱氨酸蛋白酶，可以降解a ？然而，用其特异性抑制剂胱抑素C （CysC）阻断其体内酶活性，而不是增加，降低了A？肽脑沉积在AD模型中。因为CysC蛋白（aa101- 117）的一部分，不同于它的酶抑制序列，可以结合a ？并在体外抑制原纤维的形成，我们提出在体内产生足够高水平的这种肽可能会降低A？寡聚化与神经毒性。我们展示了a)合成CysC-A？结合肽（CysC-A?）BP)特异性和优先结合纯化的A？42, b)在2个AD小鼠模型中，一个有淀粉样斑块和oA？（TgCRND8）和oA？只有（APPE693Q），它与A？c)在单次经鼻给药4天后发现了类似的共定位，d)在启动子下的cyss - gfp表达构建体经鼻给药，我们已经证明该启动子在大脑中被几种激素高度上调，产生了可检测的表达水平。我们建议：1。确定产生最佳CysC-A水平的条件？大脑中的血压。这将通过比较鼻内和鞘内递送纯化肽和表达肽的构建物，未包被或包被在微粒中，并测试哪种CysS启动子的阳性激素调节剂最能反激活CysC-A来完成。英国石油公司建造。2. 检测高水平的CysC-A？BP会降低A？-寡聚物沉积在已确诊的阿尔茨海默病中，以及在阿尔茨海默病发病前对小鼠进行慢性治疗是否会阻止其发展。被认为在A？低聚物对CysC-A？BP将用于测试这种治疗对改善学习和记忆的效果。提议用A吗？bp -纤维阻断CysC肽，而不是完整的CysC蛋白，其有害的酶抑制活性，并通过寻找新的方法来传递调节和最大化cyc - a ？BP在大脑中的表达，我们期望获得有益的效果，可能转化为人类AD的治疗和预防方法。