Development of methodologies for the analysis of DNA repair capacity to predict t

开发分析 DNA 修复能力以预测 t 的方法

• 批准号: 7682236
• 负责人: JOHN J. TURCHI
• 金额: $ 20.42万
• 依托单位: INDIANA UNIV-PURDUE UNIV AT INDIANAPOLIS
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-09-15 至 2010-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7682236
• 关键词: Adjuvant Therapy; Biological Assay; Biology; Chemotherapy-Oncologic Procedure; Cisplatin; Complex; DNA Damage; DNA Repair; DNA Repair Pathway; Development; Enzyme-Linked Immunosorbent Assay; Exposure to; Gene Expression; Goals; Human; Human Cell Line; Individual; Life; Malignant Neoplasms; Malignant neoplasm of lung; Measures; Messenger RNA; Methodology; Modeling; Modification; Molecular; Non-Small-Cell Lung Carcinoma; Nucleotide Excision Repair; Oligonucleotides; Pathway interactions; Platinum; Post-Translational Protein Processing; Proteins; Recurrence; Recurrent tumor; Repair Complex; Research; Resistance; Risk; Sampling; Smoke; Smoking; Staging; Surgical incisions; Testing; Therapeutic; Therapeutic Studies; Tissues; Tobacco smoke; base; bone; cancer diagnosis; cancer therapy; carcinogenesis; cigarette smoking; effective therapy; malignant breast neoplasm; novel; protein expression; protein protein interaction; repaired; response; tumor

DESCRIPTION (provided by applicant): Following a cancer diagnosis, determining the best course of treatment is of paramount importance. Along with recent advances in understanding the biology and pathways involved in the initiation and progression of certain cancers have come advances in individualizing treatment based on the molecular analyses of these pathways. The most convincing case involves analysis of breast cancer to determine which individuals will most likely require and benefit from adjuvant therapy. Expanding this type of analysis to other cancers holds the promise of similarly impacting cancer therapy. Considering numerous very effective therapies, including cisplatin, induce DNA damage one pathway that is directly related to how individuals respond to certain therapeutic treatments is DNA repair. In the context of cisplatin based cancer chemotherapy, reduced DNA repair capacity is associated with increased sensitivity, while increased repair activity is associated with resistance. The goal of the research described in this application is to develop methodologies to accurately determine DNA repair capacity in cancer tissue, focusing on the nucleotide excision repair (NER) pathway. The NER pathway is also responsible for removing DNA damage resulting from exposure to a variety of insults including cigarette smoke. Our hypothesis is that reduced DNA repair capacity increases the risk of smoking induced carcinogenesis and also contributes to the dramatic initial tumor regression often observed upon administration of cisplatin based therapies for treating lung cancers. The relatively short-lived response and subsequent resistance severely limits the utility of platinum based therapies. Our hypothesis is that the observed resistance is impacted by increased repair in the resistant tumors. To further test these hypotheses an accurate measure of DNA repair activity is required. Measuring gene expression or protein expression, while useful, does not always correlate with bone fide NER repair activity. Numerous NER proteins are regulated not only at the level of mRNA or protein expression, but also by posttranslational modification and protein-protein interactions. Therefore this application focuses on the development of novel methodologies to determine the extent of specific posttranslational modifications of key NER proteins and actual repair activity.

描述（由申请人提供）：在癌症诊断后，确定最佳治疗方案至关重要。沿着在理解某些癌症的起始和进展中涉及的生物学和途径方面的最新进展，在基于这些途径的分子分析的个体化治疗方面也取得了进展。最有说服力的案例涉及对乳腺癌的分析，以确定哪些个体最有可能需要并受益于辅助治疗。将这种类型的分析扩展到其他癌症，有望对癌症治疗产生类似的影响。考虑到许多非常有效的疗法，包括顺铂，诱导DNA损伤，与个体如何对某些治疗性治疗反应直接相关的一个途径是DNA修复。在基于顺铂的癌症化疗的背景下，降低的DNA修复能力与增加的敏感性相关，而增加的修复活性与抗性相关。本申请中描述的研究目标是开发准确确定癌症组织中DNA修复能力的方法，重点是核苷酸切除修复（NER）途径。NER通路也负责去除由于暴露于包括香烟烟雾在内的各种损伤而导致的DNA损伤。我们的假设是，DNA修复能力的降低增加了吸烟诱导的致癌作用的风险，并且还有助于在施用用于治疗肺癌的基于顺铂的疗法后经常观察到的显著的初始肿瘤消退。相对短暂的反应和随后的耐药性严重限制了铂类疗法的实用性。我们的假设是，观察到的耐药性受到耐药肿瘤修复增加的影响。为了进一步验证这些假设，需要精确测量DNA修复活性。测量基因表达或蛋白质表达虽然有用，但并不总是与真正的NER修复活性相关。许多NER蛋白不仅在mRNA或蛋白质表达水平上受到调节，而且还通过翻译后修饰和蛋白质-蛋白质相互作用来调节。因此，本申请的重点是开发新的方法，以确定关键NER蛋白的特定翻译后修饰的程度和实际的修复活性。