Systematic Profiling of Nonhomologous End Joining in Cancer

描述（由申请人提供）：癌症治疗通常通过使染色体断裂并超过肿瘤细胞修复它们的能力来杀死肿瘤。非同源末端连接（NHEJ）解决了大多数这样的断裂，并且已经表明癌症可能在它们执行NHEJ的程度上显著不同。 我们将确定NHEJ适应性在各种癌症中的变化程度。我们将使用特异性和敏感性定量聚合酶链反应（qPCR）测定NHEJ的效率。我们还将采用下一代测序技术来表征并行NHEJ准确性。重要的是，引入染色体断裂的癌症疗法在杀死细胞方面是有效的，因为它们引入了“脏”的断裂-断裂通常会对断裂两侧的DNA造成损伤，使其难以修复。使用具有已知NHEJ缺陷的细胞系（包括来自具有遗传性NHEJ缺陷的患者的细胞），我们已经表明，如果要以与细胞（癌症和周围正常细胞）在癌症治疗中存活的能力一致的方式评估NHEJ，则脏末端结构的使用是至关重要的。 我们将在一组癌细胞系以及原发性癌症中系统地描述NHEJ适应性。我们将确定NHEJ中癌症相关变异的程度，以及这种变异是否有助于预测治疗结果并有助于开发更好的（合理设计和靶向）治疗。

DESCRIPTION (provided by applicant): Cancer therapies often kill tumors by making chromosome breaks, and exceeding the tumor cell's ability to repair them. Nonhomologous end joining (NHEJ) resolves the majority of such breaks, and it has been suggested that cancers might differ significantly in how well they perform NHEJ. We will determine how much NHEJ fitness varies in a variety of cancers. We will measure efficiency of NHEJ with a specific and sensitive quantitative polymerase chain reaction (qPCR) assay. We will additionally employ next generation sequencing technology to characterize in parallel NHEJ accuracy. Importantly, cancer therapies that introduce chromosome breaks are effective in killing cells because they introduce breaks that are "dirty" - the break often has damage to DNA flanking the break that makes it difficult to repair. Using cell lines with known NHEJ defects (including cells from patients with inherited NHEJ defects), we have shown the employment of dirty end structures is critical if NHEJ is to evaluated in a manner consistent with the ability of cells, both cancer and surrounding normal cells, to survive cancer therapies. We will systematically profile NHEJ fitness in both a panel of cancer cell lines as well as in primary cancers. We will determine the extent of cancer-associated variation in NHEJ, and whether this variation could help predict therapy outcome and aid in development of better (rationally designed and targeted) therapies.