Alpha-Catenins in Mechanochemical Signaling

机械化学信号传导中的α-连环蛋白

• 批准号: 9130377
• 负责人: GLENN Lawrence RADICE
• 金额: $ 43.13万
• 依托单位: THOMAS JEFFERSON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-09-01 至 2017-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9130377
• 关键词: Actins; Adherens Junction; Animal Model; Atomic Force Microscopy; Attention; Binding; Biomechanics; Birth; Cadherin Domain; Cadherins; Cardiac; Cardiac Myocytes; Cardiac development; Cell Cycle; Cell Nucleus; Cells; Couples; Cytokinesis; Cytoskeletal Modeling; Cytoskeletal Proteins; Cytoskeleton; Echocardiography; Elasticity; Engineering; Exhibits; Felis catus; Generations; Genes; Genetic; Goals; Guanine Nucleotide Exchange Factors; Health; Heart; Heart failure; Homeostasis; Hydrogels; Infarction; Intercalated disc; Knock-out; Knockout Mice; Measures; Mechanics; Mediating; Membrane; Microtubules; Molecular; Mus; Muscle Cells; Myocardial Infarction; Myocardial Ischemia; Myofibrils; N-Cadherin; Nuclear; Pathway interactions; Patients; Phenotype; Phosphotransferases; Physiological; Process; Proteins; Research; Role; Signal Pathway; Signal Transduction; Stimulus; Stress; Structure; System; Tissues; Transcription Coactivator; Tubulin; Withdrawal; alpha catenin; alphaT catenin; base; cardiac regeneration; cardiac repair; cell behavior; clinical application; extracellular; heart function; improved; improved functioning; inhibitor/antagonist; injured; novel; postnatal; premature; rapid growth; response; rho; rho GTP-Binding Proteins

DESCRIPTION (provided by applicant): In the early postnatal period, cardiomyocytes undergo rapid growth and maturation that is essential for normal cardiac development. Coincident with cardiomyocyte differentiation and cell cycle withdrawal is the establishment of the N-cadherin-based adherens junction (AJ), a critical determinant of intercalated disc (ID) structure that provides the end-to-end connection between cardiomyocytes. AJ are stabilized at the ID by the dynamic I has binding of the intracellular cadherin domain to the actin cytoskeleton via beta- and alpha-catenins. To explore the role of mechanotransduction in cardiac development and homeostasis, we developed a novel animal model (alpha-cat double knockout (DKO)) in which both cytoskeletal linker proteins alphaE- and alphaT-catenin were depleted in the heart. We discovered that alpha-catenin proteins normally inhibit the ability of postnatal cardiomyocytes to re-enter the cell cycle. A significant increase in nuclear Yap, a key component of the Hippo pathway, accompanied the increased proliferation in the alpha-cat DKO hearts. Importantly, we found that alpha-cat DKO mice exhibit improved cardiac function following myocardial infarction compared to their control littermates. We hypothesize that loss of �/�catenin causes cytoskeletal rearrangement and loss of tensional stress resulting in translocation of Yap to the nucleus where it activates genes critical for cardiomyocyte proliferation. The following interrelated aims are proposed: (1) To determine the cytoskeletal-based signaling pathways that regulate the cellular activity of Yap. (2) To define the functional relationship between N-cadherin/alpha- catenin/cytoskeleton interactions and myocyte elasticity and proliferation. Deformable N-cadherin- coated hydrogels will be used to determine how stiffness and N-cadherin/catenin-mediated cytoskeletal organization cooperate to regulate proliferation. Atomic force microscopy (AFM) will be used to measure cell and tissue stiffness in alpha-cat DKO under different physiological and pathological conditions. (3) In order to pursue clinical application, we will determine the window of opportunity for manipulating alpha-catenin/Yap pathway after myocardial infarction. The long term goal of this research is to determine if interfering with the cytoskeletal proteins, alpha-catenins, represents a potential therapy to enhance cardiac repair in heart failure patients.

描述（由适用提供）：在产后早期，心肌细胞经历快速生长和成熟，这对于正常的心脏发育至关重要。与心肌细胞分化和细胞周期戒断的一致是建立了基于N-钙粘蛋白的顾问交界（AJ），这是互嵌盘（ID）结构的关键决定仪，该结构提供了心肌细胞之间的端到端连接。通过β-蛋白和α-catenins的细胞内钙粘蛋白结构域与肌动蛋白细胞骨架的动态I结合在ID处稳定。为了探索机械转导在心脏发育和稳态中的作用，我们开发了一种新型的动物模型（alpha-cat双基因敲除（DKO）），其中均在心脏中耗尽了细胞骨架接头蛋白α和α-catenin。我们发现α-钙蛋白蛋白通常会抑制产后心肌细胞重新进入细胞周期的能力。河马途径的关键组成部分的核YAP显着增加了α-cat DKO心脏增殖的增加。重要的是，我们发现与对照窝窝相比，心肌梗塞后α-Cat DKO小鼠在心肌梗塞后暴露了改善的心脏功能。我们假设损失 - /钙蛋白会导致细胞骨架重排和张力应激的丧失，从而导致YAP转移到细胞核上，在该细胞核中激活对心肌细胞增殖至关重要的基因。提出了以下相互关联的目的：（1）确定调节YAP细胞活性的基于细胞骨架的信号通路。 （2）定义N-钙粘着蛋白/α-蛋白石/细胞骨架相互作用与心肌细胞弹性和增殖之间的功能关系。可变形的N-钙粘蛋白涂层的水凝胶将用于确定刚度和N-钙粘着蛋白/catenin介导的细胞骨架组织合作如何调节增殖。原子力显微镜（AFM）将用于在不同的物理和病理条件下测量α-CAT DKO的细胞和组织刚度。 （3）为了实用临床应用，我们将确定心肌梗塞后操纵α-catenin/yap途径的机会窗口。这项研究的长期目标是确定是否干扰细胞骨架蛋白α-钙蛋白是一种潜在的疗法，以增强心力衰竭患者的心脏修复。