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，确定最好的疗程是至关重要的。随着最近在了解某些癌症发生和发展所涉及的生物学和途径方面的进展，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，导致DNA损伤的一个途径直接关系到个体对某些治疗方法的反应，那就是DNA修复。在以顺铂为基础的癌症化疗中，DNA修复能力降低与敏感性增加有关，而修复活性增加与耐药性有关。本申请中描述的研究的目标是开发方法来准确确定癌症组织的DNA修复能力，重点是核苷酸切除修复(NER)途径。NER途径还负责消除暴露在包括香烟烟雾在内的各种侮辱中造成的DNA损伤。我们的假设是，DNA修复能力的降低增加了吸烟诱发癌症的风险，也导致了肺癌治疗中经常观察到的以顺铂为基础的治疗方案的最初显著肿瘤消退。相对短暂的反应和随后的耐药性严重限制了以铂为基础的疗法的应用。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说，需要对DNA修复活动进行准确的测量。测量基因表达或蛋白质表达虽然有用，但并不总是与骨骼NER修复活动相关。许多NER蛋白不仅在mRNA或蛋白质表达水平上受到调控，而且还受到翻译后修饰和蛋白质-蛋白质相互作用的调节。因此，这项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的程度和实际的修复活动。

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.