Metals and Metal Mixtures: Cognitive Aging, Remediation and Exposure Sources (MEMCARE)

金属和金属混合物：认知老化、修复和暴露源 (MEMCARE)

• 批准号: 10580936
• 负责人: Quan Lu
• 金额: $ 1.6万
• 依托单位: HARVARD SCHOOL OF PUBLIC HEALTH
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-08-15 至 2025-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10580936
• 关键词: Age; Biological Assay; Biological Markers; Biology; California; Cell physiology; Cellular Stress; Cellular biology; Cessation of life; Chemicals; Cognition Disorders; Cognitive aging; Communication; Complex; Confocal Microscopy; Environment; Flow Cytometry; Genetic Predisposition to Disease; Goals; Home; Hydrogen Peroxide; Impaired cognition; Incidence; Inflammation; Label; Lead; Life Expectancy; Malondialdehyde; Mediating; Metal exposure; Metals; Microglia; Modeling; Molecular; Molecular Biology; Morphology; Nerve Degeneration; Neurodegenerative Disorders; Neurons; Poison; Protocols documentation; Research; Risk; Risk Factors; Ships; Signaling Molecule; Source; Stress; Structure; Superfund; Toxic effect; Toxin; Training; Universities; Work; base; early life exposure; externship; extracellular; induced pluripotent stem cell; innovation; lead exposure; molecular marker; molecular targeted therapies; neurotoxicity; novel; prevent; programs; remediation; response; sensor; therapy development

PROJECT DESCRIPTION: Increased global life expectancy has led to higher incidences of incurable neurodegenerative disorders. Although genetic predisposition and age are known risk factors for neurodegenerative disorders, it has become increasingly clear – via an unknown mechanism, exposure to lead (Pb) increases the risk of these cognitive disorders. Identifying the molecular mechanism by which early-life exposure to Pb results in neurodegeneration is essential for the development of therapies to prevent its onset. The progressive loss of structure and function in neurons which characterizes neurodegenerative disorders involves complex communication between neurons and microglia. Known for radically altering their morphology and function in response to their environmental, microglia respond to toxins, such as Pb, inducing stress and death in neighboring neurons. Using molecular and cell biology-based approaches, my current Superfund research at the MEMCARE-SRC characterizes how Pb exposure impacts signaling molecules generated by induced pluripotent stem cell (iPSC)-derived microglia. This work characterizes the extracellular signaling molecules released by microglia which potentiate neurotoxicity and may serve as biomarkers of cognitive disorders. Identifying the intracellular mechanism through which Pb alters these signaling molecules released by microglia would greatly benefit the goals of MEMCARE, which aims to evaluate how early-life exposure to metals contributes to cognitive decline. I propose to leverage a K.C. Donnelly Externship hosted in the lab of Dr. Chris Chang of the Toxic Substances in the Environment Superfund Research Program at University of California Berkeley to investigate how Pb exposure modifies microglia signaling molecules using a novel chemical-biology based approach. We hypothesize that Pb exposure increases cellular stress in iPSC-derived microglia culture. I will be trained to apply an activity-based sensing (ABS) and labeling assay to visualize and quantify two molecular markers of cellular stress in iPSC-derived microglia exposed to Pb. iPSCs will be differentiated into a stable microglia precursor in my home lab then shipped to Berkeley using an established protocol. Precursor microglia will be developed into fully matured microglia and subsequently exposed to Pb. ABS and labeling will fluorescently tag two markers of cellular stress, hydrogen peroxide and malondialdehyde, using probes developed by the Chang lab. Confocal microscopy and flow cytometry will detect and quantify these markers, respectively. Completion of this project will reveal if Pb induced cellular stress is the modifying mechanism of microglia signaling molecule release, narrowing down molecular targets for therapies aimed to prevent microglia-mediated neurotoxicity. Further, I will be trained in the use of ABS and will integrate this innovative chemical sensor with a powerful, relevant model of microglia cell function to the benefit of both Superfund centers.

项目描述： 全球预期寿命的增加导致不可治愈的神经退行性疾病的发病率增加。 虽然遗传倾向和年龄是神经退行性疾病的已知风险因素，但它已成为 越来越清楚-通过一种未知的机制，接触铅（Pb）会增加这些认知障碍的风险。 紊乱确定早期铅暴露导致神经退行性变的分子机制 对于开发预防其发作的疗法至关重要。结构和功能的逐渐丧失 在神经元中，神经退行性疾病的特征涉及神经元之间的复杂通信。 神经元和小胶质细胞。以其形态和功能的根本改变而闻名， 在环境中，小胶质细胞对毒素（如铅）作出反应，诱导邻近神经元的应激和死亡。 利用分子和细胞生物学为基础的方法，我目前在MEMCARE-SRC的超级基金研究 表征铅暴露如何影响诱导多能干细胞产生的信号分子 （iPSC）衍生的小胶质细胞。这项工作的特点是细胞外信号分子释放的小胶质细胞 其增强神经毒性并可作为认知障碍的生物标志物。识别细胞内 铅改变小胶质细胞释放的这些信号分子的机制将极大地有利于 MEMCARE的目标，旨在评估早期接触金属如何导致认知能力下降。 我建议利用一个KC Donnelly Externship在有毒物质的Chris Chang博士的实验室主持 在加州伯克利大学的环境超级基金研究项目中， 暴露使用一种新的基于化学生物学的方法改变小胶质细胞信号分子。我们 假设铅暴露增加iPSC衍生的小胶质细胞培养物中的细胞应激。我将 接受过应用基于活性的传感（ABS）和标记试验的培训，以可视化和定量两种分子 暴露于Pb的iPSC衍生的小胶质细胞中的细胞应激标志物。iPSC将分化为稳定的 我在家里的实验室里做了一个小胶质细胞前体，然后用一个既定的协议运到伯克利。前体 小胶质细胞将发育成完全成熟的小胶质细胞，随后暴露于Pb。ABS和标签将 使用探针荧光标记细胞应激的两个标志物，过氧化氢和丙二醛， 由Chang实验室开发。共聚焦显微镜和流式细胞术将检测和定量这些标志物， 分别本课题的完成将揭示铅诱导的细胞应激是否是细胞凋亡的调节机制。 小胶质细胞信号分子释放，缩小了旨在预防 小胶质细胞介导的神经毒性。此外，我将接受ABS使用方面的培训，并将这种创新的 具有强大的化学传感器，相关的小胶质细胞功能模型，以超基金都受益 中心.