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

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。随着最近对某些癌症发生和发展的生物学和途径的理解取得进展，基于这些途径的分子分析的个体化治疗也取得了进展。最具说服力的案例包括对乳腺癌的分析，以确定哪些人最有可能需要辅助治疗并从中受益。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。考虑到包括顺铂在内的许多非常有效的治疗方法，诱导DNA损伤的一个途径与个体对某些治疗方法的反应直接相关，那就是DNA修复。在以顺铂为基础的癌症化疗中，DNA修复能力的降低与敏感性的增加有关，而修复活性的增加与耐药性有关。本应用程序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，重点关注核苷酸切除修复（NER）途径。NER通路还负责清除因暴露于包括香烟烟雾在内的各种损害而造成的DNA损伤。我们的假设是，DNA修复能力的降低增加了吸烟致癌的风险，也有助于在使用以顺铂为基础的治疗肺癌的治疗中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基治疗的应用。我们的假设是，观察到的耐药性受到耐药肿瘤中修复增加的影响。为了进一步验证这些假设，需要对DNA修复活性进行精确测量。测量基因表达或蛋白表达虽然有用，但并不总是与骨内酯修复活性相关。许多NER蛋白不仅在mRNA或蛋白表达水平上受到调控，还通过翻译后修饰和蛋白-蛋白相互作用受到调控。因此，该应用侧重于开发新的方法来确定关键NER蛋白的特定翻译后修饰程度和实际修复活性。

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.