Defining Strategies to Target Energy Failure in Metabolically Vulnerable Human Cells

项目摘要 能量代谢紊乱是导致多种疾病功能障碍和死亡的主要原因，并可能 也会导致衰老。能量不足或不足以支持正常功能，可能会导致 神经退行性疾病、缺血和心力衰竭，而能量产生的不平衡可能是原因之一 致癌。因此，提高能量水平具有极大的治疗潜力，可以改善细胞功能和 生死存亡。然而，关于如何提高或保持细胞内的ATP水平，只有轶事般的例子。至 为了满足这一对抗衰老和能量集中疗法的迫切需求，我们开发了一种创新的 高通量筛选，使用荧光生物传感器测量单个细胞中的ATP水平，并使用它 确定调节三磷酸腺苷水平的基因和途径。在这个提案中，我们将检验我们的中心假设 增加三磷酸腺苷可以增强脆弱细胞的功能和存活，但效果取决于 提高三磷酸腺苷的机制。我们建议的研究的总体目标是定义最 增强细胞能量水平的强大机制，以及确定增加三磷酸腺苷是否增强功能 以及易受能源衰竭疾病影响的人类细胞的存活。我们将实现这些目标 有两个具体的目标。(1)我们将使用代谢组学、转录组学和能量产生和分析 消耗，以确定可提高能量水平的广泛机制。(2)我们会决定 如果提高三磷酸腺苷水平可以增强人神经元和心肌细胞的功能和存活。总的来说， 这些研究将i)确定可以提高细胞能量水平的广泛的细胞机制， 和二)开始评估提高能量水平以预防能量衰竭的治疗潜力 新陈代谢脆弱的人类细胞。

Project Summary Disrupted energy metabolism is a central driver of dysfunction and death in a wide range of diseases, and may also contribute to aging. Energy failure, or insufficient energy to support normal function, can lead to neurodegenerative diseases, ischemia and heart failure, while an imbalance in energy production may contribute to cancer. As such, boosting energy levels has great therapeutic potential to improve cellular function and survival. However, there are only anecdotal examples of how to increase or preserve cellular ATP levels. To address this critical unmet need for anti-aging and energy-focused therapeutics, we developed an innovative high throughput screen that uses a fluorescent biosensor to measure ATP levels in individual cells, and used it to identify genes and pathways that regulate ATP levels. In this proposal, we will test our central hypothesis that increasing ATP can enhance the function and survival of vulnerable cells, but the efficacy depends on the mechanism by which ATP is increased. The overall objectives of our proposed study are to define the most robust mechanisms to increase cellular energy levels, and to determine if increasing ATP boosts the function and survival of human cells that are susceptible to diseases of energy failure. We will accomplish these objectives in two specific aims. (1) We will use metabolomics, transcriptomics and assays of energy production and consumption to determine the broad mechanisms by which energy levels can be increased. (2) We will determine if increasing the ATP level can enhance the function and survival of human neurons and cardiomyocytes. Overall, these studies will i) determine the broad cellular mechanisms by which cellular energy levels can be increased, and ii) begin to assess the therapeutic potential of increasing energy levels to protect against energy failure in metabolically vulnerable human cells.