Defining Strategies to Target Energy Failure in Metabolically Vulnerable Human Cells

项目摘要 能量代谢紊乱是多种疾病中功能障碍和死亡的主要驱动因素， 也会导致衰老能量故障，或能量不足以支持正常功能，可能导致 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导致 到癌症因此，提高能量水平具有很大的治疗潜力，可以改善细胞功能， 生存然而，关于如何增加或保持细胞ATP水平，只有轶事的例子。到 为了解决这一关键的未满足的需求，抗衰老和能源为重点的治疗，我们开发了一种创新的 使用荧光生物传感器测量单个细胞中ATP水平的高通量筛选，并使用它 以确定调节ATP水平的基因和途径。在本提案中，我们将检验我们的中心假设 增加ATP可以增强脆弱细胞的功能和存活，但疗效取决于 ATP增加的机制。我们拟议研究的总体目标是确定最 强大的机制，以增加细胞能量水平，并确定是否增加ATP增强功能， 和人类细胞的存活，这些细胞容易受到能量衰竭的疾病的影响。我们将实现这些目标 两个具体目标。(1)我们将使用代谢组学、转录组学和能量产生测定， 能源消耗量，以确定能源水平可以提高的广泛机制。(2)我们将确定 如果增加ATP水平可以增强人类神经元和心肌细胞的功能和存活。总的来说， 这些研究将i）确定可以增加细胞能量水平的广泛的细胞机制， 和ii）开始评估增加能量水平以防止能量衰竭的治疗潜力， 代谢脆弱的人体细胞。

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.