Homeostatic Roles of Connexin43 in Response to DNA Damage

Connexin43 在 DNA 损伤反应中的稳态作用

• 批准号: 8215633
• 负责人: Daniel Sam Roh
• 金额: $ 4.72万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-03-01 至 2015-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8215633
• 关键词: Affect; Aging; Aging-Related Process; Antibodies; Apoptosis; Biological Assay; Cell Communication; Cell Proliferation; Cell Survival; Cell physiology; Cells; Cessation of life; Connexin 43; Connexins; Cornea; Corneal Endothelium; DNA Damage; DNA Repair; Data; Detergents; Deterioration; Endothelial Cells; Epithelium; Event; Exposure to; Future; Gap Junctions; Genes; Genotoxic Stress; Goals; Homeostasis; In Vitro; Intercellular Communication Inhibition; Lead; Link; Maintenance; Mediating; Modification; Molecular; Mus; Organ; Permeability; Phosphorylation; Phosphotransferases; Physiological; Proteins; Pump; RNA Interference; Resistance; Role; Signal Transduction; Site; Stress; Testing; Tissues; age related; anterior chamber; biological adaptation to stress; casein kinase; casein kinase I; cell injury; coping; coping mechanism; driving force; in vitro Model; in vivo; inhibitor/antagonist; insight; intercellular communication; lens; neuronal cell body; response; stressor; trafficking

DESCRIPTION (provided by applicant): DNA damage can significantly alter cell function and viability, thus, the accumulation of unrepaired DNA damage is believed to be a major influence in the deterioration of organ function in aging. However, cells also respond to DNA damage in many other ways beyond DNA repair in order to maintain their function and viability. In particular, genes involved in intercellular communication, such as connexins, have been identified as key components in cellular responses to numerous stressors. Despite this, the molecular mechanisms by which intercellular communication is modified after genotoxic stress have not been described. The overall goal of this F30 proposal is to understand the responses to DNA damage in corneal endothelial (CE) cells whose pump and barrier functions are essential for corneal transparency and which in vivo display age-related degeneration and accumulation of DNA damage. Using an in vitro model of CE cells, we have recently observed significant changes in the gap junction protein connexin-43 (Cx43) after exposure to DNA damage-inducing agents. To explain the cellular mechanism(s) behind these changes, we consider a potential link between DNA damage and cell communication via casein kinase-1 delta (CK1?), which is both activated by DNA damage and an essential Cx43 kinase. Our overall hypothesis is that a protective cellular response to DNA damage involves stabilization of gap junction intercellular communication which is (a) mediated by changes in Cx43 expression and site-specific phosphorylation by CK1?), and (b) required for maintenance of viability and function. This will be tested with two specific aims that will determine 1) the mechanism(s) by which DNA damage mediates modification of connexin-43 (Cx43) and 2) the physiological consequence(s) of these changes during DNA damage including effects of CE cell viability and function. The results of this project will further our understanding of stress responses to DNA damage and provide insights into how the viability of certain aging tissues may be potentially enhanced. The aging process in which the body's cells, tissues, and organs progressively deteriorate has been associated with the accumulation of DNA damage. In response to such stress, a cell may survive or perish depending on its ability to cope with that damage. This proposal studies a potential coping mechanism to DNA damage that involves changes in how damaged cells communicate with one another to maintain their function and viability.

描述（申请人提供）：DNA损伤可以显着改变细胞功能和活力，因此，未修复的DNA损伤的积累被认为是衰老过程中器官功能恶化的主要影响因素。然而，细胞还以 DNA 修复之外的许多其他方式对 DNA 损伤做出反应，以维持其功能和活力。特别是，参与细胞间通讯的基因（例如连接蛋白）已被确定为细胞对多种应激源做出反应的关键组成部分。尽管如此，基因毒性应激后细胞间通讯改变的分子机制尚未被描述。 F30 提案的总体目标是了解角膜内皮 (CE) 细胞对 DNA 损伤的反应，这些细胞的泵和屏障功能对于角膜透明度至关重要，并且在体内表现出与年龄相关的退化和 DNA 损伤的积累。使用 CE 细胞的体外模型，我们最近观察到接触 DNA 损伤诱导剂后间隙连接蛋白 connexin-43 (Cx43) 发生显着变化。为了解释这些变化背后的细胞机制，我们考虑了 DNA 损伤和通过酪蛋白激酶 1 δ (CK1?) 进行的细胞通讯之间的潜在联系，CK1 既由 DNA 损伤又由必需的 Cx43 激酶激活。我们的总体假设是，对 DNA 损伤的保护性细胞反应涉及间隙连接细胞间通讯的稳定，该通讯是 (a) 由 Cx43 表达的变化和 CK1? 的位点特异性磷酸化介导的，以及 (b) 维持活力和功能所必需的。这将通过两个具体目标进行测试，以确定 1) DNA 损伤介导 connexin-43 (Cx43) 修饰的机制，以及 2) DNA 损伤期间这些变化的生理后果，包括 CE 细胞活力和功能的影响。该项目的结果将进一步加深我们对 DNA 损伤应激反应的理解，并为如何潜在地增强某些衰老组织的活力提供见解。人体细胞、组织和器官逐渐退化的衰老过程与DNA损伤的积累有关。为了应对这种压力，细胞可能生存或死亡，这取决于它应对这种损害的能力。该提案研究了一种潜在的 DNA 损伤应对机制，涉及改变受损细胞如何相互沟通以维持其功能和活力。