Targeting tumor metabolism and immune environment via beta-catenin: Towards precision medicine in HCC

通过 β-连环蛋白靶向肿瘤代谢和免疫环境：走向 HCC 精准医疗

基本信息

批准号：
10224895
负责人：
Amaia Lujambio
金额：
$ 62.02万
依托单位：
UNIVERSITY OF PITTSBURGH AT PITTSBURGH
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/10224895
关键词：
Antigens BAY 54-9085 Biological Markers Biology CTNNB1 gene Cancer Model Cancer Patient Cells Cellular Metabolic Process Characteristics Cirrhosis Clinic Clinical Clustered Regularly Interspaced Short Palindromic Repeats Combination immunotherapy Combined Modality Therapy Data Databases Death Rate Dependence Development Enterobacteria phage P1 Cre recombinase Environment Excision FDA approved FRAP1 gene Gene Expression Gene Expression Profiling Genes Genetic Glutamate-Ammonia Ligase Glutamates Glutaminase Glutamine Hepatocyte Human Immune Immune checkpoint inhibitor Immune response Incidence Infiltration Injections Innovative Therapy Knockout Mice Lead MLL2 gene Malignant Neoplasms Malignant neoplasm of liver Medical Methionine Sulfoximine Modeling Molecular Mus Mutate Mutation Neoadjuvant Therapy Nivolumab Oncogenes Oncogenic Other Genetics PD-1 inhibitors Pathway interactions Patients Phenotype Pre-Clinical Model Precision therapeutics Predisposition Primary carcinoma of the liver cells Production Public Health Publications Publishing Regulation Reporting Research Proposals Resistance Risk Factors Role SDZ RAD SEER Program Sampling Sirolimus Sleeping Beauty Small Interfering RNA TP53 gene Tail Testing The Cancer Genome Atlas Therapeutic Transposase Tumor Immunity Unresectable Veins Woman anti-PD-1 anti-PD1 therapy base beta catenin chronic liver disease clinically relevant cohort combinatorial comparative efficacy design effective therapy gain of function mutation genome wide association study immunogenic inhibitor/antagonist lipid nanoparticle liver transplantation loss of function mutation mTOR Inhibitor mTOR inhibition men mortality mouse model mutant neoplastic cell novel overexpression palliation patient subsets personalized medicine precision medicine predicting response programmed cell death protein 1 promoter response response biomarker therapeutic target tumor tumor initiation tumor metabolism tumor-immune system interactions tumorigenesis

项目摘要

Chronic liver disease and its common sequela cirrhosis, are growing public health concerns, and major risk factors for the development of hepatocellular cancer (HCC). HCC incidence and mortality is growing in the USA. Optimal medical therapies for HCC are lacking. FDA approved agents are modestly effective. Immune checkpoint inhibitors (ICIs) including PD-1 inhibitors Nivolumab and Pembrozilumab, have been both approved and show good response rates but only in a subset of HCC cases. Biomarkers of response remain unknown. Recent years have seen a revolution in HCC GWAS studies to identify molecular drivers. Mutations in CTNNB1 activate b-catenin in the Wnt pathway & seen in up to 37% of HCCs. However, b-catenin activation alone does not lead to HCC. Analysis of 2 large HCC cohorts (TCGA & French) revealed CTNNB1 mutations to co-occur with alterations in MET, MYC, TERT, NFE2L2, MLL2, ARID2 & APOB. Overexpression/activation of Met along with CTNNB1 mutations is seen in ~11% of HCCs. Coexpression of these genes in a subset of hepatocytes using sleeping beauty transposon/transposase (SB) & hydrodynamic tail vein injection (HTVI) led to HCC by 6 weeks in mice (hMet-b-catenin model). Gene expression analysis confirmed 70% similarity between hMet-b-catenin model and HCC patient subset with Met activation & CTNNB1 mutations. In aim 1, we will generate and characterize mouse models using SB/Crispr and HTVI to co-express mutant CTNNB1 and other genes frequently co-altered in subsets of human HCC including MYC, TERT, NFE2L2, MLL2, ARID2, & APOB. We already show HCC development in Met-b-catenin, MYC-b-catenin, TERT-b-catenin & NFE2L2-b- catenin, while others are ongoing. Comparison of gene expression between mouse models and human HCC subsets will validate the relevance of these models justifying a more comprehensive cellular & molecular characterization for innovative therapies. We will also test dependence of all mutant CTNNB1-mouse models to b-catenin by using of lipid nanoparticles (LNP) containing CTNNB1-siRNA (CTNNB1-LNP) to suppress b- catenin and assess response as we have shown for Kras-b-catenin (akin to hMet-b-catenin) model. In aim 2, we will focus on b-catenin-glutamine synthetase (GS)-glutamine-mTORC1 axis in mutant-CTNNB1 HCC, recently discovered and reported by us (Publication in Cell Metabolism). All mutant-CTNNB1 driven HCC models with all co-occurrences will be tested for response to mTORC1 inhibitors like Everolimus and RM-006 (novel exclusive mTORC1 inhibitor) and to upstream GS via genetic deletion of GS in established HCCs or via use of irreversible GS inhibitor L-methionine sulfoximine (MSO) and glutaminase inhibitor CB-839 that hampers production of glutamate, a substrate for GS to generate glutamine. In aim 3, we will investigate how to make b- catenin-active HCCs shown by us to be resistant to ICIs (publication in Cancer Discovery), sensitive to ICIs through use of combination therapy. Using an immunogenic Myc-lucOS-mutant-b-catenin HCC model, that is resistant to PD-1 inhibitor, we will test role of concomitant b-catenin suppression via CTNNB1-LNP, mTORC1 inhibition via Everolimus or GS inhibition via MSO, as sensitizing agents to PD-1 inhibitor. Thus overall, our proposal will develop clinically relevant and validated preclinical models utilizing mutant b-catenin as one cooperating oncogene and demonstrate role of mutant b-catenin in regulating tumor metabolism and immune microenvironment. Completion of ours study will thus provide justification for targeting b-catenin in a notable subset of HCC patients through innovative therapies and eventually pave the way for personalized medicine.

慢性肝病及其常见后遗症肝硬化是日益增长的公共卫生问题和主要风险肝细胞癌（HCC）发展的因素。肝癌的发病率和死亡率在不断上升美国。缺乏针对 HCC 的最佳药物治疗。 FDA 批准的药物效果有限。免疫检查点抑制剂 (ICIs) 包括 PD-1 抑制剂 Nivolumab 和 Pembrozilumab，已获得批准并显示出良好的缓解率，但仅适用于一小部分 HCC 病例。反应的生物标志物仍然存在未知。近年来，HCC GWAS 研究在识别分子驱动因素方面发生了一场革命。突变 CTNNB1 激活 Wnt 通路中的 b-连环蛋白，在高达 37% 的 HCC 中可见到。然而，b-连环蛋白激活单独使用不会导致 HCC。对 2 个大型 HCC 队列（TCGA 和 French）的分析显示 CTNNB1 突变与 MET、MYC、TERT、NFE2L2、MLL2、ARID2 和 APOB 的改变同时发生。过度表达/激活 Met 与 CTNNB1 突变一起出现在约 11% 的 HCC 中。这些基因在一个子集中的共表达使用睡美人转座子/转座酶（SB）和水动力尾静脉注射（HTVI）主导的肝细胞小鼠体内 6 周后罹患 HCC（hMet-b-catenin 模型）。基因表达分析证实 70% 相似性 hMet-b-连环蛋白模型和具有 Met 激活和 CTNNB1 突变的 HCC 患者子集之间的关系。在目标 1 中，我们将使用 SB/Crispr 和 HTVI 来生成和表征小鼠模型，以共表达突变体 CTNNB1 和其他基因在人类 HCC 亚群中经常发生共同改变，包括 MYC、TERT、NFE2L2、MLL2、ARID2 等 APOB。我们已经展示了 Met-b-catenin、MYC-b-catenin、TERT-b-catenin 和 NFE2L2-b- 中 HCC 的发展连环蛋白，而其他的正在进行中。小鼠模型和人类 HCC 基因表达的比较子集将验证这些模型的相关性，证明更全面的细胞和分子创新疗法的表征。我们还将测试所有突变 CTNNB1 小鼠模型的依赖性 b-连环蛋白通过使用含有 CTNNB1-siRNA (CTNNB1-LNP) 的脂质纳米粒子 (LNP) 来抑制 b- 连环蛋白并评估反应，正如我们在 Kras-b-连环蛋白（类似于 hMet-b-连环蛋白）模型中所示的那样。在目标 2 中，我们最近将重点关注突变型 CTNNB1 HCC 中的 b-连环蛋白-谷氨酰胺合成酶 (GS)-谷氨酰胺-mTORC1 轴由我们发现并报道（发表于细胞代谢）。所有突变 CTNNB1 驱动的 HCC 模型所有同时发生的事件都将测试对 mTORC1 抑制剂（如依维莫司和 RM-006）的反应（新型独家 mTORC1 抑制剂），并通过已建立的 HCC 中 GS 的基因删除或通过使用上游 GS 不可逆的 GS 抑制剂 L-甲硫氨酸亚砜亚胺 (MSO) 和谷氨酰胺酶抑制剂 CB-839 会阻碍谷氨酸的生产，谷氨酸是 GS 生成谷氨酰胺的底物。在目标 3 中，我们将研究如何使 b- 我们显示连环蛋白活性 HCC 对 ICI 具有抵抗力（发表于 Cancer Discovery），对 ICI 敏感通过使用联合疗法。使用免疫原性 Myc-lucOS-mutant-b-catenin HCC 模型，即对 PD-1 抑制剂耐药，我们将通过 CTNNB1-LNP、mTORC1 测试伴随的 β-连环蛋白抑制的作用通过依维莫司抑制或通过 MSO 抑制 GS，作为 PD-1 抑制剂的敏化剂。因此总体而言，我们的该提案将利用突变b-连环蛋白作为一种开发临床相关且经过验证的临床前模型合作癌基因并证明突变b-连环蛋白在调节肿瘤代谢和免疫中的作用微环境。因此，我们的研究的完成将为在一个值得注意的领域中靶向 β-连环蛋白提供理由。通过创新疗法治疗 HCC 患者的子集，并最终为个性化医疗铺平道路。