Validation of WEE1 kinase as a clinical target in KRAS-mutant pancreatic cancer

胰腺导管腺癌（PDAC）是美国癌症相关死亡率的第三大原因。 States. PDAC是最致命的癌症，5年生存率只有8%。超过95%的PDAC港口 突变激活的KRAS癌基因，PDAC生长的充分验证的驱动因素。尽管几乎所有的 PDAC对致癌KRAS的依赖性，开发临床有效的PDAC抗KRAS疗法 仍然难以捉摸一种有希望的方法涉及抑制KRAS效应物信号传导，特别是KRAS抑制剂。 RAF-MEK-ERK丝裂原活化蛋白激酶（MAPK）级联。但是，我假设， 激酶也必须有助于KRAS驱动的PDAC生长。为了解决这种可能性，我们应用了一个无偏的 全激酶组化学蛋白质组学策略（MIB-MS）来鉴定新型KRAS调节激酶。急性时 KRAS抑制，我们确定了上调和下调的激酶。将此数据集与我的 分析RNA-Seq数据集，分析识别DNA损伤的ERK依赖性转录组 根据DDR和细胞周期调控基因的特征，我选择了WEE 1 DNA损伤检查点激酶 做进一步评估我的初步结果确定，突变的KRAS可能通过以下途径调节WEE 1活性： 基因转录和增加的蛋白质稳定性。我还发现对WEE 1的药理学抑制 在体外抑制PDAC生长，但在WEE 1抑制后另外观察到ERK活化。猜测 这种ERK激活是一种可以驱动耐药性的补偿机制，我发现， ERK和WEE 1抑制在体外协同抑制PDAC生长。这些结果为 我的研究和我的两个目标的基本原理：（1）描述KRAS调节的机制 WEE 1表达/丰度，并确定WEE 1缺失如何促进KRAS依赖性PDAC生长; 和，（2）应用先进的PDAC（类器官和同源原位肿瘤）临床前模型来评估 临床候选WEE 1抑制剂阿达沃塞替单独或与 临床候选ERK抑制剂ulixertinitib。我研究的科学创新包括利用DNA 检查点抑制剂作为KRAS突变型胰腺癌的治疗弱点， 护理仍然无效的常规细胞毒性药物。我的研究将进一步阐明 KRAS信号传导，并另外确定用于临床评价的新疗法。这项研究将为我提供 在癌细胞和分子生物学，临床前药物评估和实施培训， 生物信息学分析的计算工具。我还将参加指导临床活动，重点是 照顾各种各样的肿瘤病人。总的来说，该研究所的上级研究和临床机会 位于查佩尔山的北卡罗来纳州大学，我在Lineberger的经验丰富的合作者网络 综合癌症中心，以及Channing Der博士的卓越指导将推动我并为我做好准备。 成为癌症生物学领域的顶尖医生兼科学家

Pancreatic ductal adenocarcinoma (PDAC) is the 3rd leading cause of cancer-related mortality in the United States. PDAC is the deadliest cancer, with an abysmal 5-year survival rate of 8%. Over 95% of PDAC harbor a mutationally activated KRAS oncogene, a well-validated driver of PDAC growth. Despite the near-universal dependence of PDAC on oncogenic KRAS, the development of clinically effective anti-KRAS therapies for PDAC remains elusive. One promising approach involves inhibition of KRAS effector signaling, particularly inhibitors of the RAF-MEK-ERK mitogen-activated protein kinase (MAPK) cascade. However, I hypothesize that additional kinases must also contribute to KRAS-driven PDAC growth. To address this possibility, we applied an unbiased kinome-wide chemical proteomics strategy (MIB-MS) to identify novel KRAS-regulated kinases. Upon acute KRAS suppression, we identified both upregulated and downregulated kinases. Partnering this dataset with my analyses of an RNA-Seq dataset profiling the ERK-dependent transcriptome that identified DNA-damage response (DDR) and cell-cycle regulation gene signatures, I selected the WEE1 DNA damage checkpoint kinase for further evaluation. My preliminary results determined that mutant KRAS may regulate WEE1 activity through both gene transcription and increased protein stability. I also found that pharmacologic inhibition of WEE1 suppressed PDAC growth in vitro, but additionally observed ERK activation upon WEE1 inhibition. Speculating that this ERK activation is a compensatory mechanism that can drive drug resistance, I found that concurrent ERK and WEE1 inhibition synergistically suppressed PDAC growth in vitro. These results provide the foundation for my studies and the rationale for my two aims: (1) to delineate the mechanisms by which KRAS regulates WEE1 expression/abundance and determine how WEE1 loss contributes to KRAS-dependent PDAC growth; and, (2) to apply advanced preclinical models of PDAC (organoids and syngeneic orthotopic tumors) to assess the therapeutic impact of the clinical candidate WEE1 inhibitor adavosertib, alone or in combination with the clinical candidate ERK inhibitor ulixertinitib. The scientific innovation of my studies involves exploiting DNA checkpoint inhibitors as a therapeutic vulnerability of KRAS-mutant pancreatic cancer, where current standard of care remains ineffective conventional cytotoxic drugs. My studies will further elucidate basic mechanisms of KRAS signaling and additionally identify new therapies for clinical evaluation. This research will provide me with training in cancer cell and molecular biology, preclinical drug assessment, and the implementation of computational tools for bioinformatics analyses. I will also participate in mentored clinical activities focused on the care of a diverse set of oncology patients. Overall, the superior research and clinical opportunities at the University of North Carolina at Chapel Hill, my network of experienced collaborators within the Lineberger Comprehensive Cancer Center, and the exceptional mentoring of Dr. Channing Der will propel and prepare me to become a leading physician-scientist in cancer biology.