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）：核中的代谢信号传导和能量环境对于损伤后的细胞分裂和组织再生的开始至关重要。但是，控制心肌细胞周期的核ATP依赖性过程和代谢信号通路的能源供应途径尚不清楚。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化的传播中起主要作用，从线粒体到核向核以支持核能运输。使用遗传和siRNA方法的初步研究表明，AK同工型网络对于代谢重编程促进干细胞心脏分化至关重要。 W发现，在细胞周期中，胞质AK1转移到核并与有丝分裂纺锤体相关，以提供细胞分裂的能量。然而，在有丝分裂阻碍的成年心肌细胞中，AK1转移到核并未发生。此外，我们发现局部在线粒体中的AK2同工型的缺乏会阻止干细胞开发的编程，并且胚胎具有致命性。使用18o标记的技术，我们证明了心脏重新产生的能力取决于AK2的表达和通过AK-AMP-AMPK轴的AMP信号的动力学，这是p53/p21/cyclin代谢检查点调节G1/S蜂窝周期过渡的一部分。这突出了AK同工型和AMP信号网络在调节核能和细胞周期中的重要性。然而，AK转移到细胞核的分子机制以及与有丝分裂的纺锤体和细胞因子设备的关联，以及AK和AMP信号在细胞周期，心肌细胞更新和心脏再生中的重要性是未知的。目的/假设：基于新发现，我们将检验假设，即AK同工型和AMP信号的核转运对于心肌细胞周期的能量学至关重要，并且AK-AMP-AMPK信号轴是G1/S代谢检查点许可型心肌细胞的关键部分。具体目的将确定：目标1 AK同工型在心肌细胞核能和细胞周期机制的能量支持和AMP信号依赖的代谢检查点调节心脏再生潜力的重要性。 AIM 2分子机制的细胞周期依赖性AK同工型与细胞核的转运，并与有丝分裂的纺锤体和细胞因子机械以及在线粒体和核能过程的整合中的作用。 AIM 3的AIM 3机制调节AK同工型表达，代谢和生长因子通过代谢和生长因子的AMP信号传导，以促进核能和代谢检查点促进干细胞心脏分化以及心脏再生所需的成人心肌细胞周期。该应用的预期结果和新颖性将是定义首次控制核能和核能信号电路的分子机制，而细胞周期和细胞分化至关重要，这对于心脏病生成，心脏更新和再生至关重要。