Developing new conditional models to study tauopathy, amyloidosis, and their interaction

该提案的MPI独立共同开发，并彻底描述了一些最重要的 阿尔茨海默病及相关疾病（ADRD）研究常用的小鼠模型;然而，所有 现有的tau和/或淀粉样蛋白小鼠模型仍然具有限制其效用的缺点， 可以用来回答。例如，rTg 4510模型，其通过表达人P301 L tau蛋白， 强力霉素阻遏系统是该领域的黄金标准模型之一;然而，rTg 4510是有限的，部分地， 由于其依赖于两个不相关的转基因，tau蛋白病和认知功能障碍的早期发作， tau表达的泄漏。该提案的总体目标是为全球化开发新的模式， 阿尔茨海默病领域，克服了现有模型的缺点，最终提供了一个 创新的平台，其中淀粉样变性和tau蛋白病的连续性和分子生物学特性， 它们之间相互作用的基本途径可以以一种简化的、具有成本效益的方式加以审查。下 目的1，我们提出了一个模型，其中CamKII-四环素反式激活基因转基因和tau蛋白， 将共注射tau的条件表达所需的应答转基因（WT或P301 L）， 从而共整合到鼠基因组中，其随后可以作为单个等位基因传递。我们将努力 开发P301 L tau/tTA模型，其将在12-15个月时发展缠结前病理学，并在18个月时发展缠结 年龄。这种新的模型随后将充分表征生化，病理，认知和 在结构上使用MRI。一旦建立，这些新的、有条件的tau转基因将提供一种更便宜的， 更容易获得的模型，在中年到晚年发展tau蛋白病;使这两项研究能够加速 以及减缓/消除tau蛋白病。我们还预计，这种模型，如JNPL 3和rTg 4510 tau 模型，将发展神经炎症和继发性TDP-43蛋白质病。根据目标2，我们建议 通过共注射/共整合，使用cumate-阻遏物， 系统来控制APPswe/ind表达。没有小鼠模型利用大脑中的cumate抑制系统 并且简单地将APP转基因置于该可选条件系统下， 研究确定APP和tau之间的相互作用，突触核蛋白或其他潜在的相互作用物。最终在 目的3，我们将杂交新的，单等位基因，四环素抑制的tau模型与单等位基因，cumate- 抑制性APP，以允许在一个特定的细胞中解剖tau（tau蛋白病）和APP（淀粉样变性）之间的相互作用。 顺序和系统的方式。这种创新的模式，需要一个具有成本效益的，单一的育种，允许 独立控制tau和APP转基因，并将填补该领域的关键知识空白 AD竞技场中现有的动物模型目前不允许。

The MPIs for this proposal independently co-developed and thoroughly characterized some of the most commonly used mouse models for research on Alzheimer’s disease and related disorders (ADRD); however, all existing tau and/or amyloid mouse models still have shortcomings which limit their utility and the questions that they can be used to answer. For example, the rTg4510 model which expresses human P301L tau through a doxycline-repressible system is one of the gold standard models in the field; however, rTg4510 is limited, in part, by its dependence on two unlinked transgenes, the early onset of tauopathy and cognitive dysfunction, and the leakiness of the tau expression. The overall goal of this proposal is to develop new models for the Alzheimer’s Disease field that overcome the shortcomings of existing models, ultimately providing an innovative platform in which the sequential nature of amyloidosis and tauopathy and the molecular pathways underlying their interaction can be examined in a streamlined, cost-effective manner. Under Aim 1, we propose to generate a model in which the CamKII-tetracycline transactivator transgene and the tau responder transgene (either WT or P301L) required for the conditional expression of tau will be co-injected and thus co-integrated into the murine genome which can subsequently transmit as a single allele. We will strive to develop a P301L tau/tTA model that will develop pre-tangle pathology at 12-15 months and tangles at 18 months of age. This new model will subsequently be fully characterized biochemically, pathologically, cognitively and structurally using MRI. Once established, these novel, conditional tau transgenics will provide a less expensive, more accessible model that develops tauopathy in mid to late life; enabling both studies aimed at accelerating and at slowing/abrogating the tauopathy. We also anticipate that this model, like the JNPL3 and rTg4510 tau models, will develop neuroinflammation and secondary TDP-43 proteinopathy. Under Aim 2, we propose to create a new conditional APP transgenic model through co-injection/co-integration using the cumate-repressible system to control APPswe/ind expression. No mouse model utilizes the cumate-repressible system in the brain and simply having the APP transgene under this alternatively conditional system positions this model for use in studies to identify interactions between APP and tau, ?-synuclein or other potential interactors. Finally, under Aim 3, we will crossbreed the new, single allele, tetracycline-repressible tau model with the single allele, cumate- repressible APP to allow the dissection of the interaction between tau (tauopathy) and APP (amyloidosis) in a sequential and systematic fashion. This innovative model, requiring a cost-effective, single breeding, permits independent control of both the tau and APP transgenes and will position the field to fill critical gaps in knowledge that no existing animal model in the AD arena currently allows.