Understanding Compartmentalized Leucine Metabolism Downstream of mTORC1 Signaling

了解 mTORC1 信号下游的区室化亮氨酸代谢

• 批准号: 10642819
• 负责人: Paul Rosen
• 金额: $ 4.77万
• 依托单位: MASSACHUSETTS INSTITUTE OF TECHNOLOGY
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-07-01 至 2024-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10642819
• 关键词: Acetyl Coenzyme A; Address; Affect; Amino Acids; Automobile Driving; Biochemical; Biology; Biosensor; Catabolism; Cells; Cellular Metabolic Process; Communication; Complex; Critical Thinking; Cytosol; Dedications; Development; Diabetes Mellitus; Disease; Engineering; Environment; Enzymes; Epilepsy; Essential Amino Acids; FRAP1 gene; Genetic; Growth; Growth Factor; Human; Isotope Labeling; Isotopes; Leucine; Lysosomes; Malignant Neoplasms; Mass Spectrum Analysis; Measures; Metabolic; Metabolism; Methods; Mitochondria; Nutrient; Oral; Output; Pathology; Pathway interactions; Phosphotransferases; Process; Protein Biosynthesis; Protein Engineering; Proteins; Proteolysis; Reagent; Regulation; Repression; Resources; Role; Signal Transduction; Site; Starvation; Stress; Testing; Time; Training; Work; Writing; amino acid metabolism; biophysical properties; cell growth; design; experience; fluorescence imaging; inhibitor; innovation; insight; interdisciplinary approach; interest; metabolomics; novel; novel therapeutic intervention; prototype; response; science education; sensor; skills; tool

Project Summary/Abstract: The mechanistic target of rapamycin complex 1 (mTORC1) kinase coordinates cell growth and metabolism, integrating signals from growth factors, nutrients, and stresses. Many diseases, including cancers, diabetes, and epilepsy, have been associated with aberrations in mTORC1 signaling. Understanding the inputs for and outputs of mTORC1 signaling is of critical importance in driving innovations in new therapeutic interventions for these diseases. Recently, it has been appreciated that mTORC1 signaling is important for compartmentalized amino acid metabolism; this is particularly true with respect to the essential amino acid leucine. While leucine activates mTORC1 in the cytosol, mTORC1 also regulates leucine metabolism in the cytosol, mitochondria, and lysosomes. Recent work from our lab has demonstrated a key role for mTORC1 signaling in regulating the efflux of leucine and other essential amino acids from lysosomes as part of the cellular response to starvation, but how mTORC1 signaling affects leucine levels in the cytosol and mitochondria, the two other important sites of leucine metabolism in cells, remains unknown. Answering this question could help to illuminate how mTORC1 rewires amino acid metabolism in response to starvation. We will determine how mTORC1 signaling affects compartmentalized leucine metabolism through the following specific aims: 1) optimize methods to measure compartmentalized leucine levels, 2) determine how mTORC1 activity changes leucine levels in the cytosol and mitochondria, and 3) characterize how mTORC1 activity affects the rate of leucine catabolism. We will use a multidisciplinary approach, drawing on tools including genetically encoded fluorescent biosensors, organellar metabolomics, and isotope tracing, to systematically address how mTORC1 signaling affects leucine levels in the cytosol and mitochondria. Our results will be important for understanding the role of mTORC1 in regulating adaptive responses to starvation, and may identify previously unappreciated metabolic vulnerabilities in diseases with dysregulated mTORC1 signaling. The proposed work, which will take place at the MIT Biology Department/Whitehead Institute— outstanding intellectual environments that are dedicated to scientific training, includes a strong and comprehensive training plan. This plan emphasizes the development of experimental skills in cutting-edge biochemical approaches including protein engineering, fluorescence imaging, and metabolomics; critical thinking abilities; oral and written scientific communication skills; and science education experience.

项目概要/摘要： 雷帕霉素复合物1（mTORC 1）激酶的机制靶点协调细胞生长， 代谢，整合来自生长因子，营养素和压力的信号。许多疾病，包括癌症， 糖尿病和癫痫与mTORC 1信号传导的异常有关。理解输入 mTORC 1信号传导的产生和输出对于推动新治疗药物的创新至关重要。 对这些疾病的干预。最近，已经认识到mTORC 1信号传导对于 区室化的氨基酸代谢;这对于必需氨基酸尤其如此 亮氨酸。虽然亮氨酸激活胞质溶胶中的mTORC 1，但mTORC 1也调节胞质溶胶中的亮氨酸代谢。 细胞质、线粒体和溶酶体。我们实验室最近的工作证明了mTORC 1的关键作用 信号在调节亮氨酸和其他必需氨基酸从溶酶体流出的一部分， 细胞对饥饿的反应，但mTORC 1信号如何影响胞质溶胶中的亮氨酸水平， 线粒体是细胞中亮氨酸代谢的另外两个重要位点，仍然是未知的。回答这个 这个问题可能有助于阐明mTORC 1如何重新连接氨基酸代谢以应对饥饿。我们 将确定mTORC 1信号传导如何通过以下方式影响区室化亮氨酸代谢 具体目标：1）优化测量区室化亮氨酸水平的方法，2）确定mTORC 1 活性改变胞质溶胶和线粒体中的亮氨酸水平，以及3）表征mTORC 1活性如何 影响亮氨酸催化剂的速率。我们将使用多学科方法，利用工具，包括 基因编码的荧光生物传感器，细胞器代谢组学和同位素示踪，以系统地 解决mTORC 1信号如何影响细胞质和线粒体中的亮氨酸水平。我们的结果将是 对于理解mTORC 1在调节饥饿适应性反应中的作用很重要， 鉴定先前未被重视的mTORC 1信号失调疾病的代谢脆弱性。 拟议的工作将在麻省理工学院生物系/怀特黑德研究所进行- 杰出的智力环境，致力于科学训练，包括一个强大的， 全面的培训计划。该计划强调尖端实验技能的发展 生物化学方法，包括蛋白质工程，荧光成像和代谢组学;关键 思维能力;口头和书面科学沟通技巧;和科学教育经验。