Generation of a Transgenic Mouse Model for Functional Study of a Novel Gene FG01

描述（由申请人提供）：阿尔茨海默病（AD）是最常见的神经退行性疾病，影响10%的65岁以上人群和50%的85岁以上人群。神经元外斑块由？-淀粉样蛋白（A？）肽和由微管相关tau蛋白的过度磷酸化扭曲丝组成的细胞内神经元缠结（NFT）是AD的两个主要病理学标志。多线的证据表明，生产过剩/聚集的A？脑中的tau蛋白是AD发病机制的罪魁祸首，而在多种神经退行性疾病中发现了过度磷酸化的tau蛋白。因此，识别的因素参与调制的A？产生和tau蛋白过度磷酸化可能对疾病干预至关重要。使用随机纯合基因扰动（RHGP）技术，我们已经确定了一个新的基因，fg 01，并发现FG 01蛋白的过表达可以抑制A？生成和tau磷酸化。此外，我们已经证明了潜在的机制：即，FG 01可以与腺苷酸环化酶相互作用，促进cAMP合成，上调PKA活性磷酸化和磷酸化GSK-3，这反过来又影响A？生成和tau磷酸化。我们现在正在培育大脑中特异性过表达FG 01的转基因小鼠。为了进一步研究FG 01在体内的功能，我们提出了以下目标：（1）表征脑特异性FG 01转基因小鼠;（2）将FG 01转基因小鼠与3XTg-AD小鼠杂交，检测FG 01降低A？在体内，本发明的组合物还可用于抑制AD的产生、tau磷酸化和PKA和GSK-3的活性，以及减轻其它AD样表型/异常。这项工作将提供主要的洞察新基因FG 01参与AD致病事件的时间和空间进展，并可能阐明新的治疗靶点。公共卫生相关性：AD病理学的两个主要标志是细胞外淀粉样斑块和神经元内神经纤维缠结（NFT），其主要成分是A？和过度磷酸化的tau蛋白。鉴定和表征的基因参与A？产生和tau磷酸化对于理解AD发病机制至关重要。我们最近发现了一个新的基因，fg 01，其过表达已被发现减少A2的生产，tau蛋白磷酸化，GSK-3的活性，通过调节腺苷酸环化酶和PKA活性在体外。PKA和GSK-3除了在AD发病机制中起重要作用外，还对多种病理/生理过程如记忆/认知、糖尿病和肿瘤发生至关重要。因此，开发脑内过表达FG 01的转基因小鼠，并在体内研究FG 01的生理功能，将为AD的病理过程提供重要的见解。此外，FG 01转基因小鼠模型将有潜力开发新的诊断和治疗方法，用于多种疾病，包括AD，糖尿病和癌症。

DESCRIPTION (provided by applicant): Alzheimer's disease (AD) is the most common neurodegenerative disorder, affecting 10% of people over the age of 65 and 50% of the population over the age of 85. Extraneuronal plaques consisting of ?-amyloid (A?) peptides and intracellular neurofibrillary tangles (NFTs) composed of hyperphosphorylated twisted filaments of the microtubule-associated tau protein are two major pathological hallmarks of AD. Multiple lines of evidence suggest that overproduction/aggregation of A? in the brain is the prime culprit for AD pathogenesis, while hyperphosphorylated tau has been found in multiple neurodegenerative diseases. Therefore, identification of factors involved in modulation of A? generation and tau hyperphosphorylation may be crucial for disease intervention. Using the Random Homozygous Gene Perturbation (RHGP) technology, we have identified a novel gene, fg01, and found that overexpression of the FG01 protein can inhibit both A? generation and tau phosphorylation. Furthermore, we have demonstrated the underlying mechanism: i.e., FG01 can interact with adenylate cyclases to facilitate cAMP synthesis, upregulating PKA activity to phosphorylate and inactivate GSK-3, which in turn affects A? generation and tau phosphorylation. We are now generating transgenic mice that specifically overexpress FG01 in the brain. To further investigate the functions of FG01 in vivo, we propose the following aims: (1) to characterize brain-specific FG01 transgenic mice; and (2) to cross FG01 transgenic mice with 3XTg-AD mice and examine the effects of FG01 on reducing A? generation, tau phosphorylation and the activities of PKA and GSK-3 in vivo, as well as on alleviating other AD- like phenotypes/abnormalities. This work will provide major insight into the involvement of the novel gene FG01 in the temporal and spatial progression of AD pathogenic events and may potentially elucidate new therapeutic targets. PUBLIC HEALTH RELEVANCE: The two major hallmarks of AD pathologies are extracellular amyloid plaques and intraneuronal neurofibrillary tangles (NFTs), whose major components are A? and hyperphosphorylated tau, respectively. Identification and characterization of genes involved in A? generation and tau phosphorylation are critical for understanding AD pathogenesis. We have recently identified a novel gene, fg01, whose overexpression has been found to reduce A2 production, tau phosphorylation, and GSK-3 activity through modulating adenylate cyclase and PKA activity in vitro. In addition to being important to AD pathogenesis, both PKA and GSK-3 are critical for multiple patho/physiological processes such as memory/cognition, diabetes and tumorigenesis. Therefore, developing transgenic mice that overexpress FG01 in the brain and studying the physiological functions of FG01 in vivo will provide important insights into AD pathological processes. Moreover, the FG01 transgenic mouse model will have potential in developing novel diagnostic and therapeutic methods for multiple diseases including AD, diabetes and cancers.