Adenylate Kinase in Energetics of Cell Nucleus and Heart Regeneration

腺苷酸激酶在细胞核能量学和心脏再生中的作用

• 批准号: 8725218
• 负责人: PETRAS P DZEJA
• 金额: $ 38.96万
• 依托单位: MAYO CLINIC ROCHESTER
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-09-30 至 2017-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8725218
• 关键词: Adult; Area; Bioenergetics; Biology; Birth; Cardiac; Cardiac Myocytes; Cardiovascular system; Cell Cycle; Cell Differentiation process; Cell Energetics; Cell Nucleus; Cell division; Clinic; Coupling; Cyclins; Cytokinesis; Data; Development; Energy Supply; Energy Transfer; Environment; Erythrocytes; Foundations; Genetic; Growth Factor; Heart; Image; Immunochemistry; Injury; Label; Licensing; Medicine; Metabolic; Metabolism; MicroRNAs; Microtubules; Mitochondria; Mitotic spindle; Molecular; Natural regeneration; Nuclear; Nuclear Translocation; Outcome; Pathway interactions; Play; Process; Protein Isoforms; Proteomics; Psyche structure; Regenerative Medicine; Regulation; Role; Route; Scanning; Signal Transduction; Small Interfering RNA; Stem cells; Supporting Cell; System; Technology; Testing; Time; Tubulin; adenylate kinase; base; cardiac regeneration; cardiogenesis; injured; innovation; metabolomics; method development; nucleocytoplasmic transport; programs; public health relevance; regenerative; repaired; signal processing; stable isotope; stem cell differentiation; tissue regeneration; transmission process

DESCRIPTION (provided by applicant): Metabolic signaling and energetic environment in the nucleus is critical for cell division and initiation of tissue regeneration after injury. However energy supply routes to nuclear ATP-dependent processes and metabolic signaling circuits that govern cardiomyocyte cell cycle are unknown. Our studies demonstrate that adenylate kinase (AK)-phosphotransfer (2ADP<->ATP+AMP) plays a major role in metabolic signaling and transmission of high-energy phosphoryls from mitochondria to the nucleus to support nuclear transport. Preliminary studies using genetic and siRNA approaches indicate that the AK isoform network is critical for metabolic reprogram- ming facilitating stem cell cardiac differentiation. W have discovered that during cell cycle cytosolic AK1 trans- locates to the nucleus and associates with mitotic spindles to provide energy for cell division. However, AK1 translocation to the nucleus doesn't occur in mitotically arrested adult cardiomyocytes. Furthermore, we have discovered that deficiency of the AK2 isoform, which is localized in mitochondria, arrests stem cell develop- mental programming and is embryonically lethal. Using 18O-labeling technology we demonstrate that heart re-generative capacity depends on AK2 expression and dynamics of AMP-signaling through AK-AMP-AMPK axis which is a part of p53/p21/cyclin metabolic checkpoint regulating G1/S cell cycle transition. This highlights the significance of AK isoform and AMP-signaling network in regulating nuclear energetics and cell cycle. However molecular mechanisms of AK translocation to the nucleus and association with mitotic spindle and cytokinesis apparatus and the significance of AK and AMP-signaling in energy support of cell cycle, cardiomyocyte renewal and heart regeneration are unknown. Objective/Hypothesis: Based on new discoveries we will test hypothesis that nuclear translocation of AK isoforms and AMP-signaling is critical for the energetics of the cardiomyocyte cell cycle, and that AK-AMP-AMPK signaling axis is a key part of G1/S metabolic checkpoint licensing cardiomyocyte renewal and heart regeneration. The Specific Aims will determine: Aim #1 The significance of the AK isoforms in cardiomyocyte nuclear energetics and energy support of cell cycle machinery and AMP-signaling dependent metabolic checkpoint regulating heart regenerative potential. Aim #2 Molecular mechanisms of cell cycle dependent translocation of AK isoforms to the cell nucleus and association with mitotic spindles and cytokinesis machinery and the role in integration of mitochondrial and nuclear energetic processes. Aim #3 Mechanisms regulating AK isoform expression, cytosolic-nuclear distribution and AMP signaling by metabolic and growth factors in order to promote nuclear energetics and metabolic checkpoint facilitating stem cell cardiac differentiation and adult cardiomyocyte cell cycle required for heart regeneration. The expected outcome and the novelty of this application will be in defining for the first time molecular mechanisms governing nuclear energetics and AMP-signaling circuits during cell cycle and cell differentiation critical for cardiogenesis, heart renewal and regeneration.

描述（由申请人提供）：细胞核中的代谢信号和能量环境对于细胞分裂和损伤后组织再生的启动至关重要。然而，核 ATP 依赖性过程和控制心肌细胞周期的代谢信号回路的能量供应途径尚不清楚。我们的研究表明，腺苷酸激酶 (AK) 磷酸转移 (2ADP<->ATP+AMP) 在代谢信号传导和高能磷酸基从线粒体到细胞核的传输以支持核运输中发挥着重要作用。使用遗传和 siRNA 方法的初步研究表明 AK 同工型网络对于促进干细胞心脏分化的代谢重编程至关重要。 W 发现，在细胞周期期间，胞质 AK1 易位到细胞核并与有丝分裂纺锤体结合，为细胞分裂提供能量。然而，AK1 易位至细胞核不会发生在有丝分裂停滞的成年心肌细胞中。此外，我们还发现，位于线粒体中的 AK2 同工型的缺陷会阻碍干细胞的发育编程，并且具有胚胎致死性。使用 18O 标记技术，我们证明心脏再生能力取决于 AK2 表达和通过 AK-AMP-AMPK 轴的 AMP 信号传导动力学，AK-AMP-AMPK 轴是调节 G1/S 细胞周期转换的 p53/p21/细胞周期蛋白代谢检查点的一部分。这凸显了 AK 亚型和 AMP 信号网络在调节核能量和细胞周期中的重要性。然而，AK 易位至细胞核的分子机制以及与有丝分裂纺锤体和胞质分裂装置的关联以及 AK 和 AMP 信号在细胞周期、心肌细胞更新和心脏再生的能量支持中的重要性尚不清楚。目的/假设：基于新发现，我们将检验以下假设：AK 亚型和 AMP 信号传导的核转位对于心肌细胞周期的能量至关重要，并且 AK-AMP-AMPK 信号传导轴是 G1/S 的关键部分代谢检查点许可心肌细胞更新和心脏再生。具体目标将确定： 目标 #1 AK 同工型在心肌细胞核能量学和细胞周期机制的能量支持以及调节心脏再生潜力的 AMP 信号依赖代谢检查点中的重要性。目标 #2 细胞周期依赖性 AK 异构体易位至细胞核的分子机制，与有丝分裂纺锤体和胞质分裂机制的关联，以及在线粒体和核能量过程整合中的作用。目标#3 通过代谢和生长因子调节 AK 同工型表达、胞质核分布和 AMP 信号传导的机制，以促进核能量学和代谢检查点，促进干细胞心脏分化和心脏再生所需的成体心肌细胞细胞周期。该应用的预期结果和新颖性将首次定义在细胞周期和细胞分化过程中控制核能量和 AMP 信号回路的分子机制，这对心脏发生、心脏更新和再生至关重要。