Defining the Mitotic Role of Chk2 for the Treatment of Pancreatic Cancer

项目总结 基因组不稳定(GIN)是癌症的一个标志，也是治疗抵抗的主要原因。这是特别的 对于胰腺导管腺癌(PDAC)来说是真的，估计95%的PDAC细胞呈GIN。杜松子酒是 其特点是未能在间期准确修复DNA，且染色体错误率高 有丝分裂中的分离。目前还不完全清楚这些通路是如何在整个细胞周期中协同工作的 防御杜松子酒。我们以前的工作证明了DNA损伤反应(DDR)之间的联系。 蛋白质ATR和促进染色体忠实分离的有丝分裂机制。这一观察结果已经 促使我假设DDR的其他成分促进了忠诚的染色体 分离和基因组稳定性。 我的初步工作阐明了Chk2的一种新的有丝分裂功能，Chk2是一种重要的DDR激酶，被激活 在DNA双链断裂之后。我已经证明了Chk2以一种不依赖DNA损伤的方式活跃 在有丝分裂中。此外，我还证明了Chk2促进了主要有丝分裂激酶PLK1的适当活性。 在这个应用中，我建议研究chk2促进忠实染色体的机制。 分离(目标1)，确定Chk2活性如何在有丝分裂中调节(目标2)，并使用PLK1抑制 专门针对Chk2缺失的癌细胞(目标3)。我提议的研究将定义有丝分裂和dna。 Chk2的损害独立作用。这将进一步加深我们对DDR途径和有丝分裂如何 机器协同工作，以促进忠实的染色体分离和基因组稳定。最后， 根据这一提议进行的实验可能会开发出一种有针对性的治疗策略，专门杀死Chk2基因缺陷 癌症。这一点至关重要，因为Chk2功能丧失突变构成了家族性遗传性疾病的很大一部分 PDAC病例和Chk2突变使细胞对当前的一线治疗不敏感。 我的职业目标是在一家领先的机构获得一个研究教员的职位，在那里我将剖析非正统的 对DDR蛋白促进基因组稳定性的机制研究不足。我的科学发展将 成功完成这个项目，包括获得癌症生物学和老鼠方面的专业知识，这将得到支持 模特们。我将利用这些习得的技能来研究PLK1抑制剂是否可以被用来特异性地靶向 缺乏Chk2的癌细胞。重要的是，这项申请的资金也将加强我的科学培训 沟通、写作、教学、指导和职业发展，所有这些都是 成功地运营了自己的实验室。这个项目得到了一个杰出的赞助商网络的支持 将确保此项目成功完成的合作者。获得这一奖项将使我能够扩大 我的研究计划并确立了我在癌症生物学和基因组领域的主要研究人员的地位 稳定性。

PROJECT SUMMARY Genome instability (GIN) is a hallmark of cancer and a leading cause of therapeutic resistance. This is especially true for pancreatic ductal adenocarcinoma (PDAC), where an estimated 95% of PDAC cells exhibit GIN. GIN is characterized by both a failure to accurately repair DNA in interphase and high rates of chromosome mis- segregation in mitosis. It is not fully understood how these pathways work in concert throughout the cell cycle to defend against GIN. Our previous work has demonstrated a link between the DNA damage response (DDR) protein ATR and the mitotic machinery in promoting faithful chromosome segregation. This observation has prompted me to hypothesize that other components of the DDR promote faithful chromosome segregation and genome stability. My preliminary work has elucidated a novel mitotic function for Chk2, an important DDR kinase that is activated following DNA double-stranded breaks. I have shown that Chk2 is active in a DNA damage-independent manner in mitosis. Moreover, I have demonstrated that Chk2 promotes proper activity of the major mitotic kinase PLK1. In this application, I propose to investigate the mechanism by which Chk2 promotes faithful chromosome segregation (Aim 1), determine how Chk2 activity is regulated in mitosis (Aim 2), and use PLK1 inhibition to specifically target Chk2-deficient cancer cells (Aim 3). My proposed studies will define the mitotic and DNA damage-independent role for Chk2. This will further our understanding of how the DDR pathway and mitotic machinery work in concert to promote faithful chromosome segregation and genome stability. Lastly, experiments from this proposal may develop a targeted therapeutic strategy to specifically kill Chk2-deficient cancers. This is critically important, as Chk2 loss-of-function mutations make up a significant portion of familial PDAC cases, and Chk2 mutations render cells insensitive to current front-line treatments. My career goal is to obtain a research faculty position at a leading institution where I will dissect the noncanonical and understudied mechanisms by which DDR proteins promote genome stability. My scientific development will be bolstered by successful completion of this project, including gaining expertise in cancer biology and mouse models. I will use these acquired skills to investigate whether PLK1 inhibitors can be used to specifically target Chk2-deficient cancer cells. Importantly, funding for this application will also enhance my training in science communication, writing, teaching, mentoring, and career development, all of which will be essential for successfully running my own laboratory. This project is supported by an outstanding network of sponsors and collaborators that will ensure this project is successfully completed. Receipt of this award will allow me to expand my research plan and establish myself as a primary investigator in the field of cancer biology and genome stability.