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：迈向 HCC 精准医疗

• 批准号: 10605197
• 负责人: Amaia Lujambio
• 金额: $ 60.78万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 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/10605197
• 关键词: Antigens; BAY 54-9085; Biological Markers; Biology; CTNNB1 gene; Cancer Model; Cancer Patient; 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; Injections; Innovative Therapy; Knockout Mice; 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; beta catenin; chronic liver disease; clinically relevant; cohort; combinatorial; comparative efficacy; design; effective therapy; gain of function mutation; genome wide association study; immune cell infiltrate; immunogenic; inducible Cre; inhibitor; 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

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.

慢性肝病及其常见的后遗症肝硬变，是日益增长的公共卫生问题，也是主要风险 肝细胞癌(HCC)发生的因素。在中国，肝癌的发病率和死亡率正在上升。 美国。目前还缺乏治疗肝细胞癌的最佳药物。FDA批准的药物是适度有效的。免疫 检查点抑制剂(ICIS)包括PD-1抑制剂Nivolumab和Pembrozilumab，两者都是 获得批准并显示良好的应答率，但仅在部分肝细胞癌病例中。反应的生物标记物仍然存在 未知。近年来，在确定分子驱动因素的研究中，发生了一场革命。突变 在CTNNB1中，激活Wnt途径中的b-catenin&在高达37%的肝癌中可见。然而，b-连环蛋白的激活 单枪匹马不会导致肝细胞癌。对2个大型肝癌队列(TCGA和法国)的分析显示CTNNB1突变为 与MET、MYC、TERT、NFE2L2、MLL2、ARID2和APOB改变同时发生。过度表达/激活 MET伴随CTNNB1突变在约11%的肝癌中可见。这些基因在一个子集中的共表达 应用睡美人转座子/转座酶(SB)和流体动力尾静脉注射(HTVI)引导的肝细胞 HMET-b-catenin模型小鼠6周后形成肝癌模型。基因表达分析证实了70%的相似性 HMET-b-catenin模型与Met激活及CTNNB1突变的肝细胞癌患者亚群之间的关系在目标1中， 我们将使用SB/Crispr和HTVI共表达突变的CTNNB1和HTVI来建立和鉴定小鼠模型 其他在人肝癌亚群中频繁共改变的基因包括MYC、TERT、NFE2L2、ML12、ARID2、& 载脂蛋白B。我们已经展示了在Met-b-连环蛋白、MYC-b-连环蛋白、TERT-b-连环蛋白和NFE2L2-b- 连锁素，而其他的正在进行中。小鼠肝癌模型与人肝癌模型基因表达的比较 子集将验证这些模型的相关性，以证明更全面的细胞和分子 创新疗法的特征。我们还将测试所有突变的CTNNB1小鼠模型对 含CTNNB1-siRNA的脂质纳米粒(CTNNB1-LNP)抑制b-连环蛋白的研究 并评估反应，如我们对Kras-b-catenin(类似于HMET-b-catenin)模型所显示的那样。在目标2中，我们 最近将重点研究突变型CTNNB1肝癌中的b-连环素-谷氨酰胺合成酶(GS)-谷氨酰胺-mTORC1轴 由我们发现并报道(发表在《细胞代谢》杂志上)。所有突变型CTNNB1驱动的肝癌模型 将测试所有同时出现的病例对像Everolimus和RM-006(新型)这样的mTORC1抑制剂的反应 独有的mTORC1抑制剂)，并通过已建立的HCCs中GS的遗传缺失或通过使用 不可逆GS抑制剂L-蛋氨酸亚磺胺和谷氨酰胺酶抑制剂CB-839 谷氨酸的生产，谷氨酸是GS产生谷氨酰胺的底物。在目标3中，我们将研究如何使b- 我们显示连环素活性的肝癌对ICIS耐药(发表在癌症发现杂志上)，对ICIS敏感 通过使用综合疗法。使用免疫原性Myc-Lucos-突变体-b-catenin肝癌模型，即 对PD-1抑制剂的耐药性，我们将通过CTNNB1-LNP，mTORC1来测试伴随的b-连环蛋白抑制的作用 通过Everolimus抑制或通过MSO抑制GS，作为PD-1抑制剂的增敏剂。因此，总的来说，我们的 该提案将利用突变的b-连环蛋白开发临床相关和经过验证的临床前模型。 协同癌基因及突变型b-catenin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 通过创新疗法治疗肝癌患者子群，最终为个性化医疗铺平道路。

项目成果

Dual β-Catenin and γ-Catenin Loss in Hepatocytes Impacts Their Polarity through Altered Transforming Growth Factor-β and Hepatocyte Nuclear Factor 4α Signaling.

肝细胞中双 β-连环蛋白和 γ-连环蛋白丢失通过改变转化生长因子-β 和肝细胞核因子 4α 信号传导影响其极性。

• DOI: 10.1016/j.ajpath.2021.02.008
• 发表时间: 2021
• 期刊: The American journal of pathology
• 作者: Pradhan-Sundd,Tirthadipa;Liu,Silvia;Singh,Sucha;Poddar,Minakshi;Ko,Sungjin;Bell,Aaron;Franks,Jonathan;Huck,Ian;Stolz,Donna;Apte,Udayan;Ranganathan,Sarangarajan;Nejak-Bowen,Kari;Monga,SatdarshanP
• 通讯作者: Monga,SatdarshanP

Tumor-Intrinsic Mechanisms Regulating Immune Exclusion in Liver Cancers.

• DOI: 10.3389/fimmu.2021.642958
• 发表时间: 2021
• 期刊: Frontiers in immunology
• 影响因子: 7.3
• 作者: Lindblad KE;Ruiz de Galarreta M;Lujambio A
• 通讯作者: Lujambio A

β-Catenin-NF-κB-CFTR interactions in cholangiocytes regulate inflammation and fibrosis during ductular reaction.

胆管细胞中的β-catenin-NF-κB-CFTR相互作用调节导管反应过程中的炎症和纤维化。

• DOI: 10.7554/elife.71310
• 发表时间: 2021-10-05
• 期刊: eLife
• 影响因子: 7.7
• 作者: Hu S;Russell JO;Liu S;Cao C;McGaughey J;Rai R;Kosar K;Tao J;Hurley E;Poddar M;Singh S;Bell A;Shin D;Raeman R;Singhi AD;Nejak-Bowen K;Ko S;Monga SP
• 通讯作者: Monga SP

Single-cell spatial transcriptomics reveals a dynamic control of metabolic zonation and liver regeneration by endothelial cell Wnt2 and Wnt9b.

• DOI: 10.1016/j.xcrm.2022.100754
• 发表时间: 2022-10-18
• 期刊: CELL REPORTS MEDICINE
• 影响因子: 14.3
• 作者: Hu, Shikai;Liu, Silvia;Bian, Yu;Poddar, Minakshi;Singh, Sucha;Cao, Catherine;McGaughey, Jackson;Bell, Aaron;Blazer, Levi L.;Adams, Jarret J.;Sidhu, Sachdev S.;Angers, Stephane;Monga, Satdarshan P.
• 通讯作者: Monga, Satdarshan P.

Hepatocyte β-catenin loss is compensated by Insulin-mTORC1 activation to promote liver regeneration.

肝细胞β-连环蛋白损失通过胰岛素-mTORC1 激活来补偿，以促进肝再生。

• DOI: 10.1002/hep.32680
• 发表时间: 2023
• 期刊: Hepatology (Baltimore, Md.)
• 作者: Hu,Shikai;Cao,Catherine;Poddar,Minakshi;Delgado,Evan;Singh,Sucha;Singh-Varma,Anya;Stolz,DonnaBeer;Bell,Aaron;Monga,SatdarshanP
• 通讯作者: Monga,SatdarshanP