REST and Neural Network Dysfunction in Alzheimer's Disease

本RO 1赠款提案是对FOA：PAR-19-070，NOT-AG-19-033“选择性细胞”通知的回应 和衰老和阿尔茨海默病中的网络脆弱性”。阿尔茨海默病（AD）的记忆丧失 反映了神经网络功能的逐渐失效。这个提案的总体目标是探索一部小说 神经网络和REST转录因子在脑老化和AD中的作用的范例。我们有 研究表明，REST在衰老的人类大脑中被激活，但在AD中则没有，在AD中，它从 轻度认知障碍此外，我们最近证明，神经网络的调节 REST的活动参与了长寿和认知保护的保守机制。我们 初步研究表明，REST功能丧失会导致认知能力下降， 淀粉样蛋白和tau病理学。然而，还有待确定的是，REST 选择性地影响关键的神经回路或更全面地发挥作用。我们已经生成了大脑条件floxed REST敲除小鼠，现在提出产生REST敲除小鼠，选择性靶向内嗅 皮质神经元、CA 1或CA 3海马神经元或抑制性GABA能中间神经元。这些新颖 条件性REST敲除系，连同两个已建立的AD转基因系，将用于询问 REST在内嗅（EC）-海马回路中的作用，该回路在记忆形成中起重要作用 并且是AD发病的关键。具体来说，目标REST删除对神经网络的选择性影响 兴奋、突触可塑性、记忆、淀粉样蛋白沉积和tau病理学的传播将是 评估。我们的工作假设是，休息将发挥核心作用，在调节EC-海马 网络稳态，网络功能的崩溃是AD的早期组成部分。以补充 在这些体内研究中，我们建立了散发性AD和APOE 4的脑类器官模型，以询问 神经网络函数来源于散发性AD和APOE 4基因编辑的iPS细胞的脑类器官将 检查改变的神经网络活动和tau传播，以及REST的作用。无创 将在脑类器官中探索电刺激作为改善网络的干预 功能障碍最后，我们将采用一个强大的新平台，用于单细胞RNA-seq和ATAC-seq， 个脑袋这种方法将应用于小鼠模型和人类大脑，以发现基因网络， 防止年龄和AD相关的记忆丧失。这种多学科方法将涉及合作 研究人员拥有从基因组生物学到电生理学和神经病理学的专业知识， 协调努力，以促进对AD中选择性神经网络功能障碍的理解。

This RO1 grant proposal is in response to FOA: PAR-19-070, Notice NOT-AG-19-033 “Selective Cell and Network Vulnerability in Aging and Alzheimer's Disease”. Memory loss in Alzheimer's disease (AD) reflects a progressive failure of neural network function. The overall goal of this proposal is to explore a novel paradigm for the role of neural networks and the REST transcription factor in brain aging and AD. We have shown that REST is activated in the aging human brain but not in AD, where it fails beginning at the stage of mild cognitive impairment. Furthermore, we have recently demonstrated that regulation of neural network activity by REST is involved in a conserved mechanism of longevity and cognitive preservation. Our preliminary studies indicate that REST loss-of- function gives rise to cognitive decline, and markedly augments amyloid and tau pathology in several AD mouse models. It remains to be determined, however, whether REST selectively affects critical neural circuits or acts more globally. We have generated brain conditional floxed REST knockout mice, and now propose to generate REST knockout mice that selectively target entorhinal cortical neurons, CA1 or CA3 hippocampal neurons, or inhibitory GABAergic interneurons. These novel conditional REST knockout lines, together with two established AD transgenic lines, will be used to interrogate the role of REST in the entorhinal (EC)-hippocampal circuit that plays an essential role in memory formation and is central to the onset of AD. Specifically, selective effects of targeted REST deletion on neural network excitation, synaptic plasticity, memory, amyloid deposition and the propagation of tau pathology will be assessed. Our working hypothesis is that REST will play a central role in the regulation of EC-hippocampal network homeostasis, and that a breakdown of network function is an early component of AD. To complement these in vivo studies, we have established a cerebral organoid model of sporadic AD and APOE4 to interrogate neural network function. Cerebral organoids derived from sporadic AD and APOE4 gene-edited iPS cells will be examined for altered neural network activity and tau propagation, and the role of REST. Non-invasive electrical stimulation will be explored in cerebral organoids as an intervention for ameliorating network dysfunction. Finally, we will employ a powerful new platform for single cell RNA-seq and ATAC-seq in the brain. This approach will be applied to mouse models and the human brain to discover gene networks that protect against age- and AD-related memory loss. This multidisciplinary approach will involve collaborating investigators with expertise ranging from genomic biology to electrophysiology and neuropathology, in a coordinated effort to advance the understanding of selective neural network dysfunction in AD.