Deciphering the crosstalk between methionine metabolism and methyltransferases in health and disease

解读健康和疾病中蛋氨酸代谢与甲基转移酶之间的串扰

• 批准号: 10703457
• 负责人: Andrey A Parkhitko
• 金额: $ 39.75万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-12 至 2027-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10703457
• 关键词: Affect; Aging; Antioxidants; Basic Science; Biological; Biological Process; Cell physiology; Development; Disease; Foundations; Genetic; Global Change; Goals; Health; Human Pathology; Knowledge; Laboratories; Link; Malignant Neoplasms; Mediating; Metabolic Pathway; Methionine; Methionine Metabolism Pathway; Methylation; Methyltransferase; Mission; Mitochondria; Modeling; Molecular; National Institute of General Medical Sciences; Nerve Degeneration; Obesity; Oxidative Stress; Pathologic; Pathway interactions; Production; Protein Biosynthesis; Proteomics; Public Health; Reaction; Research; Research Project Grants; Resistance; S-Adenosylhomocysteine; S-Adenosylmethionine; Specificity; Starvation; Stress; System; Testing; Tissues; Work; druggable target; in vivo; innovation; methyl group; novel; novel therapeutics; prospective; response; tool

PROJECT SUMMARY/ABSTRACT Methionine metabolism is a central regulator of protein synthesis, mitochondrial function, antioxidant defense, and other critical cellular processes. Tightly regulating methionine flux via the methionine metabolism pathway is essential for healthy cellular function. Not surprisingly, an imbalance in this fundamental metabolic pathway has been attributed to numerous diseases. Yet, the molecular link between alterations in methionine availability and dysregulation of downstream cellular processes remains obscure. Methionine and ATP are the sole precursors for the production of the methyl donor S-adenosylmethionine (SAM), the principal and rate- limiting methyl donor for methyltransferases (MTs), which catalyze a variety of methylation reactions via the transfer of methyl groups onto different substrates. Although reprogramming of methionine metabolism has been observed with different pathological conditions, it is not known which downstream MTs link methionine metabolism to the development of these pathological conditions and what mediates the specificity of this interaction, representing a significant knowledge gap. I hypothesize that the identification of specific MTs will reveal novel mechanisms by which methionine metabolism regulates essential cellular processes. The goal of our research is to mechanistically understand how alterations in methionine are transduced into biological effects. To accomplish this goal, my laboratory will build and sustain three research projects. Through a preliminary screen, we identified several MTs that promote resistance to starvation or oxidative stress similar to manipulations of the methionine metabolism pathway. I will test several models to determine which MTs function downstream to methionine metabolism and complete the screen of the remaining MTs (Project 1). Secondly, I will test whether the tissue-specific expression of selected MTs help explain the specificity of how global changes in methionine levels affect specific MTs using a novel tissue-specific methionine degradation system that we recently developed (Project 2). Finally, we will use an open-ended proteomics approach to identify prospective downstream targets of the identified MTs and test how these MTs affect functional responses to stress (Project 3). These platforms interdigitate but also work independently, noting that we have already identified several MTs that promote resistance to different stresses, so Projects 2 and 3 can be performed independently of Project 1. We will employ innovative approaches by combining novel genetic tools that allow us to manipulate methionine levels within specific tissues and using a state-of- the-art approach to quantify methionine fate in vivo. The proposed research is significant because it will uncover how a central metabolic pathway (methionine) controls many basic cellular processes. This basic research is likely to further identify “druggable” targets relevant to multiple human pathologies associated with reprogrammed methionine metabolism including cancer, obesity, neurodegeneration, and aging.

项目总结/摘要 甲硫氨酸代谢是蛋白质合成、线粒体功能、抗氧化 防御和其他关键的细胞过程。通过蛋氨酸代谢严格调节蛋氨酸通量 这条通路对于健康的细胞功能至关重要。毫不奇怪，这种基础代谢的不平衡 这一途径已被归因于许多疾病。然而，甲硫氨酸的改变 下游细胞过程的可用性和失调仍然不清楚。蛋氨酸和ATP是 用于生产甲基供体S-腺苷甲硫氨酸（SAM）的唯一前体，主要和速率- 甲基转移酶（MT）的限制性甲基供体，其通过甲基转移酶催化多种甲基化反应。 将甲基转移到不同的基质上。虽然蛋氨酸代谢的重编程 在不同的病理条件下观察到，不知道哪些下游MT连接蛋氨酸 代谢对这些病理条件的发展，是什么介导的特异性， 这是一个巨大的知识差距。我假设，识别特定的MT 将揭示蛋氨酸代谢调节基本细胞过程的新机制。的 我们的研究目标是从机制上了解蛋氨酸的改变是如何被转导到 生物效应。为了实现这一目标，我的实验室将建立和维持三个研究项目。 通过初步筛选，我们确定了几种促进抗饥饿或抗氧化的MT 应激类似于甲硫氨酸代谢途径的操作。我将测试几个模型来确定 所述MT在甲硫氨酸代谢下游起作用并完成剩余MT的筛选 （项目1）。其次，我将测试所选MT的组织特异性表达是否有助于解释 甲硫氨酸水平的总体变化如何影响特定MT的特异性，使用新的组织特异性 我们最近开发的蛋氨酸降解系统（项目2）。最后，我们将使用开放式 蛋白质组学方法，以确定所确定的MT的预期下游靶点，并测试这些MT如何 影响对压力的功能反应（项目3）。这些平台相互交叉，但也独立工作， 注意到我们已经确定了几个MT，促进抵抗不同的压力，所以项目2 和3可以独立于项目1执行。我们将采用创新的方法， 新的遗传工具，使我们能够操纵特定组织内的甲硫氨酸水平，并使用 现有技术的方法来定量体内甲硫氨酸的命运。这项研究意义重大，因为它将 揭示了一个中心代谢途径（蛋氨酸）如何控制许多基本的细胞过程。这一基本 研究可能会进一步确定与多种人类病理学相关的“可药物化”目标， 重编程的甲硫氨酸代谢，包括癌症、肥胖症、神经变性和衰老。