Novel Stem Cell and Mouse Models to Study Frontotemporal Dementia

描述（由申请人提供）：几种脑部疾病是由蛋白质聚集引起的;仅额颞叶痴呆症（FTD）和阿尔茨海默病就影响了近500万美国人。尽管小鼠可以模拟疾病的各个方面，但它们可能几个月甚至几年都不会生病。相反，来自这些小鼠的细胞培养系统可以在几天或几周内提供有关疾病机制的信息，从而加速疾病治疗的发现。总体假设是，产生携带导致疾病的基因突变或组合的干细胞系将为研究发病机制提供有价值的研究工具。具体而言，将研究含有中枢神经系统干细胞的神经球在体外重现亲代小鼠品系表型中的功效。微管相关蛋白tau基因突变的小鼠可准确模拟人FTD的生化、组织学和行为学方面。疾病的突出细胞表型，过度磷酸化和tau蛋白的聚集，可以用常规的蛋白质检测方法进行跟踪，并将被用作标记物，以评估神经球及其分化后代作为模型的效用。目的1的重点是确定神经球是否表现出相同的突变特异性的时间序列的tau病理观察其亲本菌株，并研究如何增殖，存活和分化的影响转基因表达。将结果与表达人编码的野生型tau基因的神经球和小鼠进行比较。由tau磷酸化事件引起的小鼠模型化疾病的一个陷阱是内源性小鼠tau可能抑制人编码的tau磷酸化。因此，目标2的重点是测试内源性小鼠tau蛋白的表达改变体内和体外人转基因编码的tau蛋白磷酸化和聚集的假设。突变型和野生型人tau转基因都将转移到小鼠tau无效背景中。将表征小鼠和神经球并与相应的小鼠tau野生型系进行比较。如果神经球重演其亲本小鼠品系的体内表型，则它们可以作为高通量测定系统来研究参与疾病发病机制的其他基因和小分子。

DESCRIPTION (provided by applicant): Several brain diseases arise from protein aggregation; fronto-temporal dementia (FTD) and Alzheimer's disease alone affect nearly 5 million Americans. Though mice model aspects of disease, they may not become ill for months or even years. Instead, cell culture systems derived from these mice may provide information about disease mechanisms in days or weeks thereby speeding discovery of disease treatment. The overall hypothesis is that generating stem cell lines carrying mutations or combinations of genes leading to diseases will provide valuable research tools to study pathogenesis. Specifically, central nervous system stem cell containing neurospheres will be investigated for their efficacy in recapitulating parent mouse strain phenotypes in vitro. Mice with mutations in the microtubule associated protein tau gene accurately model biochemical, histological, and behavioral aspects of human FTD. Prominent cellular phenotypes of disease, hyperphosphorylation and aggregation of the tau protein, can be tracked with conventional protein detection methods and will be used as markers to assess the utility of neurospheres and their differentiated progeny as models. The focus of Aim 1 is to determine if neurospheres exhibit the same mutation-specific temporal sequence of tau pathology observed in their parental strains, and investigate how proliferation, survival, and differentiation are affected by transgene expression. Results will be compared to neurospheres and mice expressing the human encoded wild type tau gene. One pitfall to mouse modeled diseases arising from tau phosphorylation events is that endogenous mouse tau may inhibit human encoded tau phosphorylation. Therefore the focus of Aim 2 is to test the hypothesis that expression of endogenous mouse tau alters human transgene-encoded tau phosphorylation and aggregation in vivo and in vitro. Both mutant and wild type human tau transgenes will be transferred to a mouse tau null background. Mice and neurospheres will be characterized and compared to corresponding mouse tau wild type lines. If neurospheres recapitulate in vivo phenotypes of their parent mouse strains, they may serve as high through-put assay systems to investigate other genes and small molecules involved in disease pathogenesis.