Apoptotic Regulation and Neuroinflammation in Alzheimer's Disease

项目摘要 在阿尔茨海默病(AD)患者中，随着时间的推移，神经元的进行性丧失会导致记忆丧失和 其他破坏性症状。阿尔茨海默病的神经元丢失和炎症与高龄密切相关， 但仍不清楚年龄是如何导致神经细胞死亡的，也不知道细胞死亡和神经炎症是如何发生的 可能有关联。这种认识差距的存在部分是因为之前对细胞凋亡的研究，即规范调控的 细胞死亡途径，Have A)缺乏一种功能分析来测量大脑中细胞的凋亡敏感性 和b)依赖于神经元或其他受AD影响的细胞类型中可能不存在的凋亡读数。这个 我的研究目的是更好地了解衰老大脑中细胞凋亡的调节，研究它们之间的联系 在衰老、阿尔茨海默病、炎症和细胞凋亡之间。具体地说，我建议使用BH3和半胱氨酸天冬氨酸酶分析(BCP)， 对细胞凋亡敏感性的功能测量，以及其他方法来调查这一假说 衰老和阿尔茨海默病中细胞凋亡的启动促进缺乏凋亡的神经元的细胞死亡和炎症 半胱氨酸酶。在目标1中，我将量化老年小鼠的凋亡启动、caspase活性和炎症信号， 测量每个人在整个生命周期中的变化，并测试bcl2家族蛋白bax在这些变化中的作用。 在目标2中，我将研究人类诱导多能干细胞(Ipscs)中的这些现象，测试它们的效果。 家族性AD突变和cGAS/STING信号通路的调控。在目标3中，我将使用BCP和 在快速尸检人脑组织中研究凋亡启动和半胱氨酸天冬氨酸酶活性的其他方法， 阿尔茨海默病患者神经细胞凋亡启动和执行的潜在差异 和健康的捐献者对照。这三个目标的实现将产生对 细胞凋亡在衰老和阿尔茨海默病中的调控，可能为治疗干预提供新的机会。 拟议奖学金的培训计划旨在为我领导自己的研究实验室做好准备。 研究衰老和神经退行性变中的细胞死亡。该计划包括以下方面的职业发展和培训 实验室方法、主题专业知识、生物信息学分析和课堂教学。这些 我选择克里斯托弗·萨罗谢克博士和马克·阿尔贝斯博士作为 导师，让我可以借鉴他们在细胞死亡和神经变性领域的专业知识。我 我从组成我的职业发展委员会的教职员工那里寻求额外的指导和支持 和科学咨询委员会，形成了强大的导师和合作者网络。该研究和 哈佛公共卫生学院的培训环境为拟议的活动提供了理想的环境， 我将利用所有这些资源来完成本文所述的研究和培训计划。

Project Summary In patients with Alzheimer’s disease (AD), the progressive loss of neurons over time leads to memory loss and other devastating symptoms. Neuronal loss and inflammation in AD are strongly correlated with advanced age, but it remains unknown how age contributes to neuronal cell death or how cell death and neuroinflammation might be linked. This knowledge gap exists in part because previous studies of apoptosis, the canonical regulated cell death pathway, have A) lacked a functional assay to measure the apoptotic sensitivity of cells in the brain and B) relied on readouts of apoptosis that may not be present in neurons or other AD-affected cell types. The goal of my research is to better understand the regulation of apoptosis in the aging brain, investigating the links between aging, AD, inflammation, and apoptosis. Specifically, I propose to use BH3 and caspase profiling (BCP), a functional measurement of apoptotic sensitivity, and other methods to investigate the hypothesis that increased initiation of apoptosis in aging and AD promotes cell death and inflammation in neurons lacking apoptotic caspases. In Aim 1, I will quantify apoptotic priming, caspase activity, and inflammatory signaling in aged mice, measuring changes in each over lifespan and testing the role of the BCL-2 family protein BAX in these changes. In Aim 2, I will investigate these phenomena in human induced pluripotent stem cells (iPSCs), testing the effects of familial AD mutations and modulation of the cGAS/STING signaling pathway. In Aim 3, I will use BCP and other methods to study apoptotic priming and caspase competence in rapid autopsy human brain tissue, investigating potential differences in initiation and execution of apoptosis in neural cells between AD patients and healthy donor controls. The completion of these three aims will generate important new insights into the regulation of apoptosis in aging and AD, potentially revealing new opportunities for therapeutic intervention. The training plan for the proposed fellowship is designed to prepare me to lead my own research laboratory studying cell death in aging and neurodegeneration. This plan includes career development and training in laboratory methodologies, subject matter expertise, bioinformatic analysis, and classroom teaching. These training opportunities are made possible by my selection of Dr. Kristopher Sarosiek and Dr. Mark Albers as mentors, allowing me to draw on their expertise in their respective fields of cell death and neurodegeneration. I have sought out additional guidance and support from the faculty comprising my Career Development Committee and Scientific Advisory Committee, forming a strong network of mentors and collaborators. The research and training environment of the Harvard School of Public Health provides an ideal setting for the proposed activities, and I will draw on all these resources to complete the research and training plans described herein.