Comparing and contrasting the biology of damage survival at a genomic level

在基因组水平上比较和对比损伤存活的生物学

• 批准号: 7940582
• 负责人: Alexander James Bishop
• 金额: $ 44.55万
• 依托单位: UNIVERSITY OF TEXAS HLTH SCIENCE CENTER
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-06-01 至 2013-09-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7940582
• 关键词: Alkylation; Animal Model; Bioinformatics; Biology; Bleomycin; Cells; Critical Pathways; DNA Damage; Data; Data Analyses; Data Set; Disease; Disease model; Drosophila genus; Embryo; Environmental Exposure; Eukaryotic Cell; Event; Exposure to; Fibroblasts; Functional disorder; Future; Gene Mutation; Genes; Genomics; Goals; Human; Individual; Individual Differences; Knowledge; Lead; Libraries; Life; Malignant Neoplasms; Mammalian Cell; Maps; Medicine; Methods; Monitor; Mus; Organism; Pathway interactions; Positioning Attribute; Predisposition; Proteins; Publications; RNA Interference; Reagent; Research; Research Infrastructure; Role; Screening Result; Technical Expertise; Testing; Therapeutic Intervention; Ultraviolet Rays; Validation; Work; base; cell injury; cost; environmental agent; fitness; gene function; genome-wide; improved; insight; interest; multicatalytic endopeptidase complex; novel; protein protein interaction; public health relevance; response; time use

DESCRIPTION (provided by applicant): Exposure to exogenous agents that damage the cell is an unavoidable and potentially deleterious component of life, though various cellular responses exist to counter these exposures. It is understood that these countermeasures are vital for both cellular and organism fitness and is key to understanding disease predisposition, individual sensitivity to environmental exposures and developing medicine. However, we still lack significant knowledge in what responses are involved in environmentally induced damage and, even for those responses we have identified, many mechanistic details are lacking. Our hypothesis is that (a) the genes and pathways necessary to survive damage can be both damage-specific and -nonspecific, (b) despite differences for individual gene requirements in surviving different damages, they may share the same basic pathways for survival and (c) we can determine novel functions for identified proteins based on interactions with proteins already known to function in damage survival. This premise is supported by our preliminary data. Our goal is to identify damage survival mechanisms in mammalian cells. Using the model organism Drosophila and RNA interference we identified genes and pathways necessary for cells to survive different DNA damaging agents, such as proteasome activity. Once these genes are determined in Drosophila cells, we can test whether the same genes contribute to survival in mammalian cells. Indeed, we already successfully used this strategy to identify novel alkylation survival genes and pathways in mouse and human cells and demonstrated that pathways critical in Drosophila are also involved in mammalian damage response. Here we propose to leverage this strategy by first completing the validations for two additional survival screens of agents that cause different forms of damage, bleomycin and ultraviolet light. In Aim 1, we prioritize the screen results, complete screen validations, and identify novel survival genes and pathways not previously known to be involved in damage survival, demonstrating the functional conservation of these pathways in mouse embryonic fibroblasts. The focus of the latter part of this aim will be on novel proteins and differences between damaging agents within established survival pathways. In Aim 2, we take a reverse strategy, identifying survival proteins that alter damage induced proteasome activity, investigate the mechanism of their interaction with proteasome and differences between damage types. Available information on established pathways and protein: protein interaction network maps will facilitate this work. We are uniquely positioned to carry out the proposed work with the necessary infrastructure, collaborators and technical expertise, as demonstrated by our publications and our preliminary data that supports our hypothesis. In summary, this research will provide insights into the complicated biology of damage response and survival, identifying novel genes that may be important for gene/environmental interactions, disease predisposition and chemo-therapeutic intervention. Future directions will be to apply the knowledge gained to pertinent disease models. PUBLIC HEALTH RELEVANCE: Environmental exposures are a normal and unavoidable consequence of life, however, the resultant cellular damages must be counteracted in order maintain cellular and organism fitness. Deficiency in a countermeasure is one basis for disease predisposition or sensitivity to particular environmental agents. Here we propose to identify novel components to these countermeasures necessary for damage survival, improve our understanding of how these and already established countermeasures interact and set the groundwork for future work to harness this knowledge to improve disease prediction and treatment.

描述（由申请人提供）：暴露于损害细胞的外源性药物是生活中不可避免的，可能是有害的组成部分，尽管存在各种细胞反应来抵抗这些暴露。可以理解，这些对策对于细胞和生物体的适应性至关重要，对于理解疾病易感性，对环境暴露和发展医学的个人敏感性是关键。但是，我们仍然缺乏在环境引起的损害中涉及哪些响应的知识，即使对于我们已经确定的那些响应，也缺乏许多机械细节。我们的假设是（a）（a）生存损害所必需的基因和途径既可以是损害特异性的，也可能是非特异性的，尽管在生存不同的损害中，单个基因需求的差异差异，但它们可能具有相同的生存途径，并且（c）我们可以基于已知的损害生存的蛋白质来确定与已知蛋白质相互作用的新功能。我们的初步数据支持此前提。我们的目标是确定哺乳动物细胞中的损伤生存机制。使用模型生物果蝇和RNA干扰，我们鉴定了细胞在不同的DNA损伤剂（例如蛋白酶体活性）中生存所需的基因和途径。一旦这些基因在果蝇细胞中确定，我们就可以测试相同的基因是否有助于哺乳动物细胞的存活。确实，我们已经成功地使用了该策略来鉴定小鼠和人类细胞中新型的烷基化存活基因和途径，并证明果蝇中至关重要的途径也参与了哺乳动物损伤的反应。 在这里，我们建议通过首先完成两个造成不同形式损害，博来霉素和紫外线的额外生存屏幕的验证来利用这种策略。在AIM 1中，我们优先考虑屏幕结果，完成屏幕验证，并确定新的生存基因和以前未知与损伤存活有关的途径，这证明了这些途径在小鼠胚胎成纤维细胞中的功能保护。该目标的后半部分的重点将放在已建立的生存途径中破坏药物之间的新型蛋白质和差异上。在AIM 2中，我们采用反向策略，识别改变损伤引起蛋白酶体活动的生存蛋白，研究其与蛋白酶体的相互作用的机制以及损伤类型之间的差异。有关既定途径和蛋白质的可用信息：蛋白质相互作用网络图将有助于这项工作。正如我们的出版物和支持我们假设的初步数据所证明的那样，我们有独特的位置，可以使用必要的基础架构，合作者和技术专长进行拟议的工作。总而言之，这项研究将为损害反应和生存的复杂生物学提供见解，确定对基因/环境相互作用，疾病易感和化学治疗干预可能很重要的新型基因。未来的方向将是将获得的知识应用于相关疾病模型。 公共卫生相关性：环境暴露是生命的正常且不可避免的后果，但是，必须抵消由此产生的细胞损害，以维持细胞和生物体的适应性。对策中的不足是疾病易感或对特定环境药物敏感性的基础。在这里，我们建议确定损害生存所必需的这些对策的新成分，以提高我们对这些和已经建立的对策如何相互作用的理解，并为将来的工作奠定基础，以利用这一知识来改善疾病的预测和治疗。