Genome Integrity Through the Cell Cycle in Homeostasis and Cancer

项目摘要/摘要 哺乳动物基因组的稳定性依赖于一套出色的监测、修复、 信令和检查点机制。DNA本身的突变，或许多 这些基因组完整性机制，可能导致包括癌症在内的疾病。给定 突变和细胞分裂在肿瘤发生中的重要性--基因组中的两条中心途径 完整性是DNA损伤反应和细胞分裂周期。一个更完整的 了解这些途径对于我们了解细胞的正常发育是至关重要的。 动态平衡、干细胞生物学、癌症病因学以及以下技术应用 基因打靶。该项目旨在解决这些领域的突出问题，以便 阐明可提高治疗效果的基本分子细胞生物学机制 癌症的结局。 F99阶段的重点是双链断裂(DSB)修复和末端控制 切除，是通过钝端连接还是通过以下方式修复DSB的关键分子“选择” 同源重组。我揭示了关于蛋白质控制的新的机制见解 这个选择，53BP1，与PARP1治疗BRCA1缺陷性癌症相关的研究结果 抑制剂。我们发现--而不是像通常认为的那样阻碍末端切除--53BP1 招募聚合酶α通过填充切除的DSB合成来抵消切除。在 在接下来的工作中，我将探索BRCA1和53BP1如何调节切除和 根据这一新模型进行填充式合成。我还将获得必要的经验和曝光率 从科学和专业上过渡到一流实验室里专注于癌症的博士后。 对于K00阶段，我将转移我的重点和方法来研究保存机制 在有丝分裂的关键窗口中的基因组完整性，其中不同的染色质生物学途径 会合。我计划学习并实现高通量筛选，计算分析 更大的、统计功能强大的数据集，以及在小鼠体内的建模，以及对 对人类肿瘤样本的数据进行测序。这些新的方法，再加上我已经 在遗传学、显微镜和生物化学方面有深厚的背景，这将使我能够解决大多数 当今基因组完整性和癌症生物学中的紧迫和具有挑战性的问题。借助于…… 此次获奖，我打算继续我的研究贡献并积累经验，以便 成为我自己的癌症实验室的领导者，也是基因组完整性领域的领导者。

Project Summary/Abstract The stability of the mammalian genome depends on a remarkable toolkit of surveillance, repair, signaling, and checkpoint mechanisms. Mutations in the DNA itself, or corruptions in many of these genome integrity mechanisms, can result in disease including cancer. Given the importance of mutation and cell division in tumorigenesis, two central pathways in genome integrity are the DNA damage response, and the cell division cycle. A more complete understanding of these pathways is crucial to our knowledge of normal cellular development in homeostasis, stem cell biology, the etiology of cancers, as well as for technical applications like gene targeting. This project seeks to tackle outstanding problems in these fields in order to elucidate fundamental molecular cell biology mechanisms that can improve therapeutic outcomes in cancer. The F99 phase is focused on double-strand break (DSB) repair and the control of end resection, a critical molecular ‘choice’ of whether to repair a DSB by blunt end-joining or by homologous recombination. I revealed novel mechanistic insights about the protein controlling this choice, 53BP1, findings relevant to the treatment of BRCA1-deficient cancers with PARP1 inhibitors. We found that—instead of blocking end resection as was generally thought—53BP1 recruits polymerase alpha to counteract resection by fill-in synthesis of the resected DSB. In the remainder of the dissertation work, I will explore how BRCA1 and 53BP1 regulate resection and fill-in synthesis in light of this new model. I will also gain the necessary experience and exposure scientifically and professionally to transition to a cancer-focused postdoc in a stellar lab. For the K00 phase, I will shift my focus and approaches to study mechanisms preserving genome integrity in the critical window of mitosis, where diverse chromatin biology pathways converge. I plan to learn and implement high throughput screens, computational analysis of larger, statistically-powerful data sets, as well as in vivo modeling in the mouse, and analysis of sequencing data from human tumor samples. These new approaches, coupled with my already strong background in genetics, microscopy, and biochemistry, will allow me to address the most pressing and challenging issues in genome integrity and cancer biology today. With the aid of this award, I intend to continue my research contribution and gain experience in order to become a leader of my own cancer-focused lab and a leader in the field of genome integrity.