STK11 loss of function and anti-PD-1 therapy resistance in KRAS-driven lung adenocarcinoma

项目总结 肺癌仍然很普遍，而且很致命，强调需要继续研究和治疗。 发展。免疫治疗技术的进展，包括检查点抑制剂，如抗PD-1 单抗治疗，在肺癌患者的治疗中显示出巨大的前景。不幸的是，并不是全部 病人会有反应。最近的临床研究表明，抗PD-1治疗耐药与特定的 体细胞肿瘤突变。值得注意的是，KRAS驱动的肺腺癌患者缺乏功能性STK11 预后更差，转移率增加，对抗PD-1治疗明显耐药。为什么 STK11功能丧失(LoF)与抗PD-1治疗耐药的相关性尚不清楚。丝氨酸苏氨酸 蛋白激酶11(STK11)编码一种重要的肿瘤抑制基因，在KRAS诱导的肺组织中频繁突变 腺癌。我们在人肺癌细胞系中的工作支持STK11缺失依赖的转录 诱导肿瘤细胞因子是可能影响抗PD-1治疗抵抗的机制之一。具体来说， RNAseq分析表明YAP1和核因子KappaB(NFkB)被激活 转录网络在STK11缺失时发生。YAP1是河马信号轴的终极效应器，一个 在发育过程中调节器官大小的关键途径。虽然YAP1之前曾与 STK11是否直接调控肿瘤细胞因子的表达，其活性是否受到STK11的直接调控，目前尚不清楚。 NFkB是调节多种细胞因子必不可少的主要转录因子，但与YAP1一样，STK11是如何 可能监管NFkB活动的问题仍未解决。我们假设STK11缺失导致抗PD-1治疗 通过改变肿瘤细胞因子改变免疫细胞对肿瘤微环境的募集 发信号。我们提出的工作计划旨在表征可诱导的不同免疫细胞的招募 小鼠肺癌模型与STK11状态和肿瘤转录组的关系。我们将利用免疫 能够产生可诱导的KRAS驱动或KRAS/STK11 LOF的合格的基因工程小鼠 直接表征肿瘤基因、肿瘤转录组和腺癌之间的相关性 不同的免疫细胞招募。然后，我们将利用这个模型来评估逆转抗PD-1病毒的策略 治疗抵抗。具体地说，我们将评估下游的拮抗候选转录网络 STK11缺失，包括YAP1和NFkB，作为恢复抗肿瘤免疫微环境的策略， 潜在挽救STK11缺失肿瘤的抗PD1疗效。为了实现这一点，我们将定义 我们的小鼠表现出依赖STK11的抗PD-1耐药性，与仅由KRAS驱动的肿瘤相比。我们 然后将评估拮抗候选网络是否逆转STK11-LoF转录表型， 并反过来恢复在STK11 WT肿瘤中观察到的免疫细胞互补。

PROJECT SUMMARY Lung cancer remains both prevalent and deadly, emphasizing the need for continued research and therapy development. Advances in immunotherapeutic technologies, including check-point inhibitors such as anti-PD-1 monoclonal antibody therapy, have shown great promise in treating lung cancer patients. Unfortunately, not all patients respond. Recent clinical studies have linked anti-PD-1 therapy resistance with specific combinations of somatic tumor mutations. Notably, patients with KRAS-driven lung adenocarcinomas lacking functional STK11 suffer a worse prognosis, increased rates of metastasis, and marked resistance to anti-PD-1 therapy. Why STK11 loss of function (LoF) correlates with resistance to anti-PD-1 therapy remains unclear. Serine Threonine Kinase 11 (STK11) encodes an important tumor suppressor gene frequently mutated in KRAS-driven lung adenocarcinomas. Our work in human lung cancer cell lines supports STK11-loss-dependent transcriptional induction of tumor cytokines as one mechanism that might impact anti-PD-1 therapy resistance. Specifically, RNAseq analysis indicates activation of Yes-associated protein 1 (YAP1) and Nuclear Factor Kappa B (NFkB) transcription networks occur upon STK11 loss. YAP1 is the ultimate effector in the HIPPO signaling axis, a pathway critical for regulating organ size during development. While YAP1 has previously been linked with regulating tumor cytokine expression, whether its activity is directly regulated by STK11 has yet to be established. NFkB is a master transcription factor essential for regulating numerous cytokines, but as with YAP1, how STK11 might regulate NFkB activity remains unaddressed. We hypothesize that STK11 loss leads to anti-PD-1 therapy resistance by altering immune cell recruitment to the tumor microenvironment via altered tumor cytokine signaling. The workplan we present aims to characterize differential immune cell recruitment in an inducible mouse model of lung cancer as a function of STK11 status and tumor transcriptome. We will leverage immune competent, genetically engineered mice that develop inducible KRAS-driven or KRAS/STK11 LoF lung adenocarcinomas to directly characterize the correlation between tumor genotype, tumor transcriptome, and differential immune cell recruitment. We will then utilize this model to assess strategies for reversing anti-PD-1 therapy resistance. Specifically, we will evaluate antagonizing candidate transcriptional networks downstream of STK11 loss, including YAP1 and NFkB, as strategies to restore an anti-tumor immune microenvironment, potentially rescuing anti-PD1 efficacy in STK11 null tumors. To accomplish this, we will define the extent of STK11-dependent anti-PD-1 resistance displayed by our mice compared with tumors driven by KRAS alone. We will then evaluate whether antagonizing candidate networks reverses the STK11-LoF transcriptional phenotype, and in turn restores the immune cell compliment observed in STK11 WT tumors.