Apoptotic Regulation and Neuroinflammation in Alzheimer's Disease

项目摘要 在阿尔茨海默病（AD）患者中，神经元随时间的进行性丧失导致记忆丧失， 其他致命症状。AD中的神经元损失和炎症与高龄密切相关， 但年龄如何导致神经细胞死亡或细胞死亡和神经炎症 可能有联系。这种知识差距的存在，部分是因为以前的研究细胞凋亡，典型的调节， 细胞死亡途径，缺乏一种功能测定来测量脑细胞的凋亡敏感性 和B）依赖于可能不存在于神经元或其他AD影响的细胞类型中的凋亡的读数。的 我的研究目标是更好地了解衰老大脑中细胞凋亡的调节，调查 衰老、AD、炎症和凋亡之间的联系。具体来说，我建议使用BH 3和半胱天冬酶谱（BCP）， 凋亡敏感性的功能性测量，以及其他方法来研究增加细胞凋亡的假设。 衰老和AD中细胞凋亡的启动促进缺乏凋亡的神经元中的细胞死亡和炎症 半胱天冬酶。在目标1中，我将量化衰老小鼠的凋亡启动、半胱天冬酶活性和炎症信号， 测量每一个在寿命期间的变化，并测试BCL-2家族蛋白BAX在这些变化中的作用。 在目标2中，我将在人类诱导多能干细胞（iPSC）中研究这些现象，测试其影响。 家族性AD突变和cGAS/STING信号通路的调节。在目标3中，我将使用BCP， 研究快速尸检人脑组织中凋亡启动和半胱天冬酶活性的其它方法， 研究AD患者神经细胞凋亡启动和执行的潜在差异 和健康供体对照。这三个目标的完成将产生重要的新的见解， 调节衰老和AD中的细胞凋亡，潜在地揭示了治疗干预的新机会。 拟议的奖学金的培训计划是为了让我做好准备，领导我自己的研究实验室 研究衰老和神经退化中的细胞死亡。该计划包括职业发展和培训， 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·萨罗西克博士和马克·阿尔伯斯博士作为我的培训对象， 导师，让我利用他们在细胞死亡和神经变性各自领域的专业知识。我 我从我的职业发展委员会成员那里寻求额外的指导和支持 和科学咨询委员会，形成了一个强大的导师和合作者网络。研究与 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.