iPSC from British and Danish dementias: new discovery tools for brain amyloidoses

描述（由申请人提供）：最近已证明将原代真皮成纤维细胞重编程为诱导多能干细胞（iPSC）有助于产生源自神经退行性疾病患者的活神经元。这项技术为创建体外模型以研究相关人类细胞类型中的疾病病理生理学提供了巨大的希望，否则将无法获得。家族性英国和丹麦痴呆（分别为FBD和FDD）是常染色体显性病症，其非常类似于阿尔茨海默病（AD）的许多临床和神经病理学特征，包括实质淀粉样蛋白和前淀粉样蛋白病变、广泛的脑淀粉样蛋白血管病和神经纤维缠结，在形态学和免疫化学上与AD中的那些难以区分。值得注意的是，从FBD沉积物-ABri-和FDD病变-ADan-中分离出的淀粉样蛋白亚基在结构上与阿尔茨海默病A？无关，这清楚地表明不同的淀粉样蛋白肽可能引发类似的神经病理学变化，导致相同的情况：CAA相关的微血管功能障碍，神经元丢失和痴呆。因此，这些家族性疾病构成了有希望的替代范例，以更好地了解淀粉样蛋白在疾病发病机制的复杂机制中的作用。鉴于AD、FBD和FDD之间的许多临床和神经病理学相似性，我们提出i）使用重复mRNA转染从FBD和FDD患者队列以及从两种疾病的非携带者同胞获得的真皮成纤维细胞产生和表征iPSC系，这是研究小组成功使用的一种更安全的非DNA整合技术;和ii）进一步区分iPSC细胞系， 新产生的iPSC转化为活的和功能性的神经元和内皮细胞，其特征在于通过良好建立的形态学、分子和生物学标准。我们预计，这些iPSC衍生的成熟细胞将成为研究与FBD和FDD疾病表型相关的特定分子和时间方面的优秀候选细胞。此外，它们将在神经退行性疾病领域产生更广泛的影响，从这些罕见疾病扩展到AD领域，为更好地理解调节APP加工的机制提供宝贵的选择，稳态和tau过度磷酸化的过程，作为高通量药物筛选平台的替代范例，并协助鉴定将CAA相关的血脑屏障功能障碍和微血管形成的发展与神经血管单位的变化和认知障碍联系起来的串扰途径。该提案代表了纽约大学医学院和加州大学欧文分校研究人员的合作努力，并建立在参与研究人员的互补专业知识及其对脑淀粉样蛋白疾病分子发病机制的长期兴趣的基础上。

DESCRIPTION (provided by applicant): Reprogramming of primary dermal fibroblasts into induced pluripotent stem cells (iPSCs) has recently proven to be instrumental for the generation of viable neurons derived from patients with neurodegenerative disorders. This technology holds tremendous promise for the creation of in vitro models to study disease pathophysiology in relevant human cell types that would otherwise be impossible to obtain. Familial British and Danish dementias (FBD and FDD, respectively) are autosomal dominant conditions that closely resemble many clinical and neuropathological features of Alzheimer's disease (AD) including parenchymal amyloid and pre- amyloid lesions, widespread cerebral amyloid angiopathy and neurofibrillary tangles morphologically and immunochemically indistinguishable from those in AD. Notably, the amyloid subunits isolated from FBD deposits -ABri- and FDD lesions -ADan- are structurally unrelated to the Alzheimer's A¿, a clear indication that different amyloid peptide could trigger similar neuropathological changes leading to the same scenario: CAA-related microvascular dysfunction, neuronal loss and dementia. Thus, these familial disorders constitute promising alternative paradigms to better understand the role of amyloid in the complex mechanisms of disease pathogenesis. In view of the many clinical and neuropathological similarities between AD, FBD and FDD, we are proposing i) to generate and characterize iPSC lines from dermal fibroblasts obtained from a cohort of FBD and FDD patients as well as from non-carrier siblings of both disorders using repetitive mRNA transfections, a safer non- DNA-integrating technology successfully used by the research team; and ii) to further differentiate the newly generated iPSCs into viable and functional neurons and endothelial cells characterized through well- established morphological, molecular and biological criteria. We anticipate that these iPSC-derived mature cells will constitute excellent candidates to study specific molecular and temporal aspects linked to FBD and FDD disease phenotypes. Moreover, they will have a broader impact in the field of neurodegenerative disorders, extending beyond these rare diseases into the field of AD, providing invaluable options for a better understanding of the mechanisms that modulate APP processing, A¿ homeostasis and the process of tau hyperphosphorylation, serving as alternative paradigms for high throughput drug screening platforms, and assisting with the identification of cross-talk pathways connecting CAA-associated blood brain barrier dysfunction and development of microhemorrhages with changes in the neurovascular unit and cognitive impairment. This proposal represents a collaborative effort from investigators of New York University School of Medicine and the University of California, Irvine and builds on the complementary expertise of the participating researchers and their long-standing interest in the molecular pathogenesis of cerebral amyloid disorders.