Targeting REST in Alzheimer's Disease

项目摘要/摘要 阿尔茨海默病(AD)的记忆丧失反映了环境中神经网络功能的进行性衰竭 病理和神经炎症。这项提议的总体目标是探索一种新的范式 REST转录因子在AD神经网络和小胶质细胞功能调节中的作用 提出一种新的治疗方法。我们发现主发育调节因子REST/NRSF是 在老化的人脑中诱导，并协调保护衰老的基因网络的表达 神经毒性应激和过度兴奋所致的神经元。REST在AD中下调，从 轻度认知障碍。我们对条件性休息缺陷小鼠的初步研究表明，休息 防止淀粉样蛋白和tau病理，以及AD小鼠模型的认知能力下降。开始，开始 了解REST在关键神经回路中的作用，我们将在 第二层内嗅皮质神经元，或在CA1或CA3海马神经元中，内嗅觉的组成部分 皮层(EC)--AD早期受影响的海马区。这些新的REST基因敲除小鼠将被杂交 用已建立的AD转基因系来探索对神经网络兴奋、突触可塑性、 记忆和病理学。我们还将扩展最近的初步研究，表明REST在小胶质细胞中丢失 AD，而休息不足的小胶质细胞功能受损。为了探索REST在小胶质细胞中的作用，我们 已经产生了一个有条件的小胶质细胞REST-基因敲除小鼠品系。此小鼠模型与主模型一起 小胶质细胞培养，将用于研究静息调节的免疫/炎症信号通路， 以及对吞噬功能和Aβ清除的影响。将进行RNA-SEQ和CHIP-SEQ分析以 广义定义REST调节的基因网络和影响小胶质细胞功能的信号通路。一个中环 问题是，REST是否是一个可以下药的目标。我们已经确定了新的激活休息的药物，这些药物可以防止 阿尔茨海默病小鼠模型的记忆丧失和病理改变。休息不足的小鼠失去了药物活性，这表明 休息的专一性。重要的是，激活静息的药物原型没有显示出毒性的证据。 慢性给药两年以上的小鼠。这些新颖的代理不仅提供了证据 翻译原则，但也是发现REST中介的潜在机制的宝贵工具 神经保护。一种这样的机制是未折叠的蛋白质反应，它可能受到休息的调节 通过靶基因和信号通路的激活将被研究。这是一个多学科的 该方法可能为衰老和AD的神经保护机制提供新的见解，具有潜在的 具有重要的治疗意义。

Project Summary/ Abstract Memory loss in Alzheimer's disease (AD) reflects a progressive failure of neural network function in the setting of pathology and neuroinflammation. The overall goal of this proposal is to explore a novel paradigm for the role of the REST transcription factor in the modulation of neural networks and microglial function in AD, and to advance a new therapeutic approach. We discovered that the master developmental regulator REST/NRSF is induced in the aging human brain and coordinates the expression of a gene network that protects aging neurons from neurotoxic stress and hyperexcitation. REST is downregulated in AD, beginning at the stage of mild cognitive impairment. Our preliminary studies in conditional REST-deficient mice indicate that REST protects against amyloid and tau pathology, as well as cognitive decline in AD mouse models. To begin to understand the role of REST in critical neural circuits, we will generate highly selective knockouts of REST in layer II entorhinal cortical neurons, or in CA1 or CA3 hippocampal neurons, components of the entorhinal cortex (EC)-hippocampal circuit that is affected early in AD. These novel REST knockout mice will be crossed with established AD transgenic lines to explore effects on neural network excitation, synaptic plasticity, memory and pathology. We will also extend recent preliminary studies showing that REST is lost in microglia in AD, and that REST-deficient microglia are functionally impaired. To explore the role of REST in microglia, we have generated a conditional microglial REST-knockout mouse line. This mouse model, together with primary microglial cell cultures, will be used to investigate REST-regulated immune/inflammatory signaling pathways, as well as effects on phagocytosis and Aβ clearance. RNA-seq and ChIP-seq analysis will be performed to broadly define REST-regulated gene networks and signaling pathways that affect microglial function. A central question is whether REST is a druggable target. We have identified novel REST-activating drugs that prevent memory loss and pathology in AD mouse models. Drug activity is lost in REST-deficient mice, indicating specificity for REST. Importantly, the prototype REST-activating drug does not show evidence of toxicity in mice following chronic administration for more than two years. These novel agents not only provide proof-of- principle for translation, but are also valuable tools for discovering underlying mechanisms of REST-mediated neuroprotection. One such mechanism is the unfolded protein response, which may be modulated by REST activation through target genes and signaling pathways that will be investigated. This multidisciplinary approach may provide novel insights into neuroprotective mechanisms in aging and AD, with potentially important therapeutic implications.