Understanding the molecular mechanism of memory from single-cell gene expression to protein folding

项目摘要/摘要 认知功能和记忆缺陷是神经退行性疾病的一个衰弱方面，导致 长期的残疾、个人及其家庭遭受的巨大痛苦以及巨大的社会经济代价。 目前，有1600多万美国人患有认知障碍，这一数字预计还会继续 随着阿尔茨海默病和相关痴呆症的患病人数预计将翻一番，人数也在上升 到2060年。长期记忆巩固需要在特定的时间段诱导基因表达 图案。新合成的转录本被翻译并折叠成功能蛋白，然后被运输到 正确的蜂窝位置。这些步骤中的任何一个步骤的功能障碍都会导致记忆障碍，并可能 在疾病条件下调节失调，尽管发生这种情况的确切机制尚不清楚。我的龙- 学期目标是确定Nr4A亚家族调节基因的特定分子机制 表达来控制长期记忆，以便开发治疗AD的工具，我将把它作为 一家专注于研究的机构的独立调查员。这项提议的总体目标是 确定记忆形成过程中转录调控和蛋白质折叠之间的联系机制 这些过程的破坏如何导致阿尔茨海默病的认知障碍。我的中央 假设是Nr4A调节特定细胞类型的基因，以促进蛋白质折叠和记忆形成。 我将通过以下三个目标来测试这一点：特定目标1(K99)：识别细胞类型特定转录 利用单细胞识别记忆巩固过程中Nr4A的记忆特征和直接基因靶点 RNA测序法；特定目标2(K99)：确定Nr4A靶标的作用，即蛋白质折叠 伴侣Hspa5，在记忆巩固中，并确定其下游蛋白靶点；特定目标3(R00)： 确定激活Nr4A转录因子对小鼠记忆缺陷和基因表达的影响 阿尔茨海默病的模型。该项目提供尖端研究技能培训，包括 记忆研究中单细胞数据和染色质浓缩的计算分析。爱荷华大学 是记忆、计算精神病学、神经退行性疾病和分子生物学专家的家园， 协作环境提供了获得必要技能的理想环境，使我能够 过渡到一个成功的独立研究生涯。因此，在指导(K99)阶段，我将 参与为我成功实现独立做好准备的活动，包括科学培训 演讲、实验室管理技能、拨款撰写工具、科学同行评议和面试/申请 准备工作。总而言之，这个奖项将为我提供分子方面的尖端技能和专业知识 生物学、神经退行性变和行为药理学，以确保强大的技术和 启动我的独立实验室调查机制的概念基础 阿尔茨海默氏症。

Project Summary / Abstract Deficits in cognitive function and memory are a debilitating aspect of neurodegenerative disease resulting in long-term disability, enormous suffering to individuals and their families, and significant socioeconomic cost. Currently, more than 16 million Americans live with cognitive impairment, and this number is expected to continue rising as the number of individuals affected by Alzheimer's disease and related dementias is predicted to double by 2060. Long-term memory consolidation requires the induction of gene expression in a specific temporal pattern. Newly synthesized transcripts are translated and folded into functional proteins and then trafficked to the correct cellular location. Dysfunction in any of these steps can lead to memory impairment and may be dysregulated in disease conditions, although the precise mechanisms by which this occurs are unclear. My long- term goal is to determine the specific molecular mechanisms through which the Nr4A sub-family regulates gene expression to control long-term memory in order to develop therapeutic tools to treat AD, which I will pursue as an independent investigator at a research-focused institution. The overall objective of this proposal is to determine the mechanisms that link transcriptional regulation and protein folding during memory formation and how the disruption of these processes contributes to cognitive impairment in Alzheimer's disease. My central hypothesis is that Nr4A regulates genes in specific cell types to facilitate protein folding and memory formation. I will test this through the following three aims: Specific Aim 1 (K99): identify cell-type specific transcriptional signatures of memory and identify direct gene targets of Nr4A during memory consolidation using a single-cell RNA sequencing approach; Specific Aim 2 (K99): determine the role of an Nr4A target, the protein folding chaperone Hspa5, in memory consolidation and identify its downstream protein targets; Specific Aim 3 (R00): determine the effect of activating Nr4A transcription factors on memory deficits and gene expression in a mouse model of Alzheimer's disease. This project provides training in cutting-edge research skills, including computational analysis of single cell data and chromatin enrichment in memory research. The University of Iowa is home to experts on memory, computational psychiatry, neurodegeneration, and molecular biology, and the collaborative environment provides an ideal setting in which to obtain the necessary skills that will allow me to transition into a successful independent research career. As such, during the mentored (K99) phase, I will engage in activities designed to prepare me to successfully achieve independence, including training in scientific presentations, laboratory management skills, grant writing tools, scientific peer-review, and interview/application preparation. Collectively, this award will provide me with the cutting-edge skills and expertise in molecular biology, neurodegeneration and behavioral pharmacology necessary to ensure a strong technical and conceptual foundation to start my independent laboratory investigating mechanisms for the treatment of Alzheimer's disease.