Characterization of YAP as a rational companion target in lung cancer

项目摘要。肺癌是世界范围内癌症死亡的主要原因，非小细胞肺癌。 肺癌(NSCLC)是肺癌的主要组织亚型，肺腺癌是肺癌的主要亚型 非小细胞肺癌。尽管靶向治疗在临床上取得了进展，但耐药性仍然是一个问题 限制了病人的生存。耐药性进化的另一个较不为人所知的方面是耐药， “持久”细胞状态，在这种状态下，一群癌细胞在最初的靶向治疗中存活下来，形成最低限度的 残留病(MRD)水库，是获得性抗性的前兆。我们提出一种创新的、 多学科和协作项目，希望通过定义 河马-YAP通路在促进药物耐受和对当前靶向通路的MRD中的作用 治疗。扩展我们在R01的第一个资金周期中完成的工作，我们的目标是利用我们的 发现河马-YAP信号通路作为许多疾病的关键分子回路和治疗靶点 RTK-RAS-MAPK信号过度激活导致的癌症。我们的数据显示了一个新兴的范式 在靶向RTK-RAS-MAPK过程中，YAP的哪个激活是耐药状态的关键功能特征 非小细胞肺癌中的抑制作用。我们观察了多癌基因驱动的NSCLC临床前治疗期间YAP的激活情况 用EGFR、ALK、KRAS和SHP2抑制剂处理的动物模型。YAP上调促进HSP70基因的表达 几种生存因子包括bclxl和RTK，如FGFR1和ErbB2。此外，通过单一- 细胞RNA测序(ScRNAseq)在治疗患者的临床标本中纵向进行 利用靶向抑制剂，我们发现残留的癌细胞通常表现出谱系可塑性和向 一种肺泡型(AT)1/2样转录状态，是一种新的慢周期耐药癌症表型 细胞。我们证实，这发生在真正的癌细胞中(即，不是来自错误注释的正常肺泡细胞)和 不同于单纯治疗的腺癌或正常的AT1/2细胞。我们的数据表明YAP在 促进药物耐受性和这种新形式的血统可塑性。YAP/TEAD对YAP信号的抑制作用 临床开发中的小分子抑制剂抑制耐药癌细胞存活和 AT1/2样谱系开关分子标记的表达。我们建议进一步检验这一假设 YAP信号是调节RTK-MAPK通路生物学和临床反应的关键分子开关 抑制剂。在目标1中，我们定义了YAP在促进药物耐受和AT1/2样谱系可塑性中的作用 存在于RTK-RAS-MAPK驱动的非小细胞肺癌耐药癌细胞中。我们还测试了药理作用是否 在临床前模型中，抑制YAP/TEAD可以抑制MRD并增强对RTK-RAS抑制剂的反应。在……里面 目的2研究YAP/TEAD激活的分子特征和AT1/2样谱系开关作为脑出血的生物标志物。 治疗前后使用患者肿瘤的MRD和临床结果。该项目提供了对 YAP和谱系可塑性在治疗耐受性和MRD中的作用，具有未来临床翻译的潜力。

PROJECT ABSTRACT. Lung cancer is the leading cause of cancer mortality worldwide, with non-small cell lung cancer (NSCLC) the predominant histologic subtype of lung cancer and lung adenocarcinoma the major subset of NSCLC. Despite clinical progress with the use of targeted therapies, drug resistance remains a problem that limits patient survival. A less well understood aspect of the evolution of drug resistance is the drug tolerant, “persister” cell state, in which a subpopulation of cancer cells survives initial targeted treatment to form a minimal residual disease (MRD) reservoir that is a precursor to acquired resistance. We propose an innovative, multidisciplinary and collaborative project to hopefully improve the survival of NSCLC patients by defining the role that the Hippo-YAP pathway plays in promoting drug tolerance and MRD to current pathway targeted therapies. Extending work that we completed in the first funding cycle of this R01, we aim to capitalize on our discovery of the Hippo-YAP signaling pathway as a critical molecular circuit and therapeutic target in the many cancers driven by hyperactivation of RTK-RAS-MAPK signaling. Our data suggest an emerging paradigm in which YAP activation is a key functional feature of the drug tolerant state during RTK-RAS-MAPK targeted inhibition in NSCLC. We observed YAP activation during therapy in multiple oncogene-driven NSCLC preclinical models treated with EGFR, ALK, KRAS and SHP2 inhibitors. YAP upregulation promoted the expression of several survival factors including BCL-xL and RTKs such as FGFR1 and ErbB2. Furthermore, through single- cell RNA sequencing (scRNAseq) performed longitudinally in clinical specimens obtained from patients treated with targeted inhibitors, we discovered that residual cancer cells often show lineage plasticity and a transition to an alveolar type (AT)1/2-like transcriptional state that is a novel phenotype of slow cycling, drug tolerant cancer cells. We verified this occurred in bona fide cancer cells (i.e., not from misannotated normal alveolar cells) and was distinct from therapy-naïve adenocarcinoma or normal AT1/2 cells. Our data suggest a role for YAP in promoting drug tolerance and this novel form of lineage plasticity. Inhibition of YAP signaling with YAP/TEAD small molecule inhibitors that are in clinical development suppressed drug tolerant cancer cell survival and expression of molecular markers of the AT1/2-like lineage switch. We propose to further test the hypothesis that YAP signaling is a key molecular switch that regulates the biological and clinical response to RTK-MAPK pathway inhibitors. In Aim 1, we define the role of YAP in promoting drug tolerance and the AT1/2-like lineage plasticity present in drug tolerant cancer cells in RTK-RAS-MAPK-driven NSCLCs. We also test whether pharmacologic inhibition of YAP/TEAD can thwart MRD and enhance response to RTK-RAS inhibitors in preclinical models. In Aim 2, we study molecular features of YAP/TEAD activation and the AT1/2-like lineage switch as biomarkers of MRD and clinical outcomes using tumors from patients before and on treatment. This project offers insight into the role of YAP and lineage plasticity in therapy tolerance and MRD, with potential for future clinical translation.