CARM1-mediated regulation of YAP1 as a therapeutic target in lung cancer

CARM1 介导的 YAP1 调节作为肺癌的治疗靶点

基本信息

批准号：
10391561
负责人：
JOSEPH KISSIL
金额：
$ 42.87万
依托单位：
H. LEE MOFFITT CANCER CTR & RES INST
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-08-01 至 2025-04-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10391561
关键词：
Adenocarcinoma Cell Affect Arginine Binding Blood Vessels Cancer Etiology Cancer Patient ChIP-seq Clinical Trials Coculture Techniques Communication Complex Cytotoxic T-Lymphocytes Dimensions Disease Down-Regulation Drug resistance Epidermal Growth Factor Receptor Epidermal Growth Factor Receptor Tyrosine Kinase Inhibitor Epithelial Erlotinib Event F-Box Proteins Gene Expression Regulation Gene Targeting Genetic Genetic Transcription Growth Hematologic Neoplasms Human Hydroxylation Hypoxia Immune checkpoint inhibitor Immunosuppression Immunotherapy In Vitro Isogenic transplantation KRAS2 gene Lead Light Lung Lung Adenocarcinoma MEKs Malignant Neoplasms Malignant neoplasm of lung Malignant neoplasm of pancreas Mediating Mediation Mesenchymal Methylation Modality Molecular Mus Mutation Nature Neoplasm Metastasis Neurofibromin 2 Non-Small-Cell Lung Carcinoma Normal tissue morphology Nuclear Hormone Receptors Oncogenic Operative Surgical Procedures Organ Size PLK1 gene Pancreas Pathway interactions Patients Phosphotransferases Platinum Compounds Primary Neoplasm Proteins RNA Processing Recurrence Recurrent tumor Regulation Reporting Resistance development Role Sampling Signal Pathway Signal Transduction Smoker Survival Rate T-Lymphocyte TANK-binding kinase 1 TBK1 gene Testing Therapeutic Time Transcription Coactivator Transcription Factor AP-1 Transferase United States Xenograft Model amlexanox angiogenesis anti-cancer anticancer activity base cell transformation chemotherapeutic agent coactivator-associated arginine methyltransferase 1 combat driver mutation experimental study gemcitabine improved in vivo inhibitor lung cancer cell mimicry mortality mouse model mutant non-smoker novel novel strategies overexpression patient subsets response self-renewal stem-like cell targeted treatment taxane therapeutic target transcriptome sequencing transplant model tumor tumor progression tumor xenograft

项目摘要

Non-small cell lung cancer (NSCLC) is the leading cause of cancer-related mortality and can occur in smokers as well as non-smokers. Components of the Hippo tumor suppressive pathway are altered in NSCLC and recent studies have shown that this pathway can be modulated by upstream signaling events from K-Ras and EGFR. We had found that YAP1 could induce Sox2 and EMT markers, promoting the growth and metastasis of NSCLC. We find that the non-canonical IkB kinase TBK1 (Tank Binding Kinase 1) could physically interact with YAP1 and phosphorylate it; further, depletion of TBK1 led to a marked elevation of YAP1 levels under normoxic conditions, but only in K-Ras mutant, but not EGFR-mutant, lung adenocarcinoma cells. This was specific to TBK1, since depletion of the closely related IKKe kinase did not elevate YAP1 levels. We find that the induction of YAP1 occurs at the level of protein stability, brought about by the methylation of arginine residues of YAP1 by CARM1 (PRMT4). CARM1 is overexpressed in a variety of cancers and high levels of CARM1 correlates with poor survival in NSCLC patients. Based on these results, we hypothesize that the regulation of YAP1 by TBK1 and CARM1 is a novel mechanism which significantly promotes the growth and metastasis of non-small cell lung cancer. Studies proposed in this application will characterize this regulation mechanistically using a variety of in vitro and in vivo analysis, including co-culture studies and syngeneic transplantation models combined with global analysis of gene regulation by arginine-methylated YAP1 protein. The physical interaction of YAP1 with TBK1, PRMT5 and CARM1 will be assessed in three different human lung cancer TMAs from low grade and high grade tumors that harbor various K-Ras mutations; such an analysis will shed light on whether the levels and physical interaction of YAP1 with these regulatory molecules affect the growth and progression of these tumors. ChiP-re-ChIP, ChIP-Seq and RNA-Seq analysis will be conducted on primary tumor samples and adjacent normal tissue to identify the downstream targets of arginine-methylated YAP1. YAP1 has been demonstrated to have significant immunosuppressive effects; downregulating YAP1 through the inhibition of CARM1 can be expected to enhance the efficacy of immune checkpoint inhibitors. In depth studies will be conducted on syngeneic mouse models to assess how CARM1 inhibitors alone or in combination, affects the anti-tumor activity of T cells. Further, the fact that CARM1 inhibitors are in clinical trials for hematological malignancies raise the possibility that their utility can eventually be extended to K-Ras mutant lung cancers. We propose to test the anti-cancer efficacy of the CARM1 inhibitor EZM2302 alone or in combination with the TBK1 inhibitor Amlexanox, the K-Ras G12C inhibitor AGM510 or the MEK inhibitor Trametinib or the PLK1/K-Ras. Given the established oncogenic role of CARM1 and YAP1 in various cancers including those of the pancreas and the lung, an elucidation of these novel regulatory modes of YAP1 function would lead to the identification of vulnerabilities that could potentially be targeted to combat K-Ras mutant lung adenocarcinomas.

非小细胞肺癌（NSCLC）是癌症相关死亡率的主要原因，可能发生在吸烟者中以及非吸烟者。 NSCLC和最近研究表明，该途径可以通过K-RAS和EGFR的上游信号事件调节。我们发现YAP1可以诱导SOX2和EMT标记，从而促进NSCLC的生长和转移。我们发现非典型的IKB激酶TBK1（储罐结合激酶1）可以与YAP1进行物理相互作用并磷酸化；此外，TBK1的耗竭导致YAP1水平在常氧化下的显着升高条件，但仅在K-RAS突变体中，但不在EGFR突变剂中，肺腺癌细胞。这是特定于 TBK1，由于密切相关的IKKE激酶的耗竭没有升高YAP1水平。我们发现归纳 Yap1的属于蛋白质稳定性的水平，由Yap1的精氨酸残基甲基化引起 CARM1（PRMT4）。 Carm1在多种癌症中过表达，高水平的Carm1与 NSCLC患者的生存率差。基于这些结果，我们假设TBK1对YAP1的调节 Carm1是一种新型机制，可显着促进非小细胞肺的生长和转移癌症。该应用中提出的研究将使用各种IN机械地表征该调节体外和体内分析，包括共培养研究和合成性移植模型精氨酸甲基化YAP1蛋白对基因调节的全球分析。 yap1与 TBK1，PRMT5和CARM1将在低级和具有各种K-RAS突变的高级肿瘤；这样的分析将阐明水平是否 YAP1与这些调节分子的物理相互作用会影响这些分子的生长和进展肿瘤。将在原发性肿瘤样品和相邻的正常组织以鉴定精氨酸 - 甲基化YAP1的下游靶标。 yap1已经证明具有明显的免疫抑制作用；通过抑制下调YAP1 可以预期CARM1可以增强免疫检查点抑制剂的功效。深度研究将是在合成小鼠模型上进行的，以评估CARM1抑制剂的单独或组合如何影响 T细胞的抗肿瘤活性。此外，CARM1抑制剂正在血液学试验中恶性肿瘤增加了其效用最终可以扩展到K-Ras突变肺癌的可能性。我们建议单独测试CARM1抑制剂EZM2302的抗癌疗效或与TBK1结合抑制剂amlexanox，K-RAS G12C抑制剂AGM510或MEK抑制剂Trametinib或PLK1/K-RAS。鉴于Carm1和Yap1在包括胰腺的各种癌症中已建立的致癌作用肺，阐明这些新型YAP1功能的调节模式将导致鉴定有可能针对K-RAS突变体肺腺癌的脆弱性。