Project 2: Synthetic Lethal Therapy for HPV-Negative Head and Neck Cancer

摘要 人乳头瘤病毒阴性的头颈部鳞状细胞癌通常会失去G1/S细胞周期检查点， 大多数肿瘤都有TP53突变，许多肿瘤还突变了其他肿瘤抑制基因，如CDKN2A。 这类肿瘤依赖于与G2/M相关的检查点来修复因 复制压力和其他基因组侮辱。这种依赖性表明对肿瘤有选择性的易感性 合成致死策略通过G2/M控制进展。在广泛的初步数据中，我们表明 G2/M检查点激酶WEE1的抑制剂AZD1775/adavosertib可有效增敏TP53mut HNSCC 对Aurora A激酶(AURKA)的抑制作用。Wee1诱导CDK1抑制Y15-磷酸化， 阻断M期进入，从而钝化AURKA抑制的细胞毒效应。Wee1抑制消除了这一点 阻止、加速携带高度破坏性纺锤体异常和其他缺陷产生的细胞的有丝分裂进入 从AURKA抑制，导致有丝分裂灾难和细胞凋亡。我们发现了AURKA/WEE1的组合 抑制作用在HNSCC异种移植中有效，而在正常组织和细胞中耐受性良好。我们已经延长了这一期限 概念，确定WEE1和其他G2/M调节激酶之间的其他有前景的药物组合 (PLK1和CHK1)。值得注意的是，在合成致命治疗中存活下来的细胞数量有限，其特点是 非整倍体和其他缺陷表明它们可能增加了肿瘤突变负担(TMB)，表达 新抗原和上调的炎症信号与免疫检查点抑制的敏感性有关。 该项目将把这一观察结果直接带到临床上。我们将进行术前窗口期I和 新一代高效选择性AURKA抑制剂LY3295668的扩大临床试验 Adavosertib组合，建立概念的药效学证据，为患者识别生物标志物 在未来的研究中进行选择，并将增效组合置于基因组改变和治疗的背景下 抵抗。在目标1中，我们将评估联合致死的机制，并使用单细胞测序和 Luminex分析用于查询TMB、预测新抗原和测量炎症信号。AIM 2将质疑 常见HNSCC TP53和CDKN2A突变类型以及顺铂耐药性对A Wee1-AURKA抑制剂组合，使用定义的细胞系模型和患者来源的异种移植(PDX)。在……里面 目的3，我们将进行术前窗口试验，以确定推荐的第二阶段剂量，确定活性 并建立概念的药效学证据，并评估推测的预测生物标志物的反应 联合应用WEE1/AURKA抑制HNSCC的作用。

Summary HPV-negative head and neck squamous cell carcinomas (HNSCC) typically lose G1/S cell cycle checkpoints, with most tumors having mutations in TP53, and many also mutating other tumor suppressors such as CDKN2A. Such tumors become dependent on checkpoints associated with G2/M to repair DNA damage arising from replication stress and other genomic insults. This dependency suggests a tumor-selective vulnerability to synthetic lethal strategies controlling progress through G2/M. In extensive preliminary data, we show that AZD1775/adavosertib, an inhibitor of the G2/M checkpoint kinase WEE1, potently sensitizes TP53mut HNSCC cell lines to inhibition of Aurora A kinase (AURKA). WEE1 induces an inhibitory Y15-phosphorylation of CDK1, blocking M-phase entry and thus blunting the cytotoxic effects AURKA inhibition. WEE1 inhibition abrogates this arrest, accelerating mitotic entry for cells bearing highly disruptive spindle abnormalities and other defects arising from AURKA inhibition, resulting in mitotic catastrophe and apoptosis. We found combined AURKA/WEE1 inhibition is potent in HNSCC xenografts, while well-tolerated in normal tissue and cells. We have extended this concept, identifying additional promising drug combinations between WEE1 and other G2/M regulatory kinases (PLK1 and CHK1). Notably, the limited number of cells surviving synthetic lethal treatment are characterized by aneuploidy and other defects suggesting they may have increased tumor mutation burden (TMB), express neoantigens, and upregulated inflammatory signaling associated with sensitivity to immune checkpoint inhibition. This project will take this observation directly to the clinic. We will conduct a pre-operative window phase I and expansion clinical trial of the late generation, high potency selective AURKA inhibitor LY3295668 with adavosertib combination, establishing pharmacodynamic proof of concept, identifying biomarkers for patient selection in future studies, and placing synergistic combinations in context of genomic alteration and treatment resistance. In Aim 1, we will evaluate mechanisms of combination lethality, and use single cell sequencing and Luminex profiling to query TMB, predict neoantigens, and measure inflammatory signaling. Aim 2 will query the effect of classes of common HNSCC TP53 and CDKN2A mutations, and cisplatin resistance, on response to a WEE1-AURKA inhibitor combination, using defined cell line models and patient derived xenografts (PDXs). In Aim 3, we will perform a pre-operative window trial to establish recommended phase 2 doses, determine activity and establish pharmacodynamic proof of concept, and to evaluate putative predictive biomarkers for response to combination WEE1/AURKA inhibition in HNSCC.