Investigating Immunosuppression in Beta-catenin-mutated Hepatocellular Carcinoma for Improved Precision Medicine Therapeutics

研究β-连环蛋白突变肝细胞癌的免疫抑制以改进精准医学治疗

• 批准号: 10749344
• 负责人: Brandon Lehrich
• 金额: $ 5.27万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-07-01 至 2026-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10749344
• 关键词: Antigen Presentation; Automobile Driving; Award; BAY 54-9085; Binding; Bioinformatics; Biological Response Modifiers; CTNNB1 gene; Cancer Biology; Cancer Etiology; Cell Proliferation; Cells; Clinical; Combined Modality Therapy; Data; Development; Disease; Education; Exclusion; Future; Gene Expression; Genes; Growth; Immune; Immune System Diseases; Immune Targeting; Immune checkpoint inhibitor; Immune response; Immunohistochemistry; Immunooncology; Immunosuppression; Immunotherapeutic agent; Immunotherapy; Incidence; Inflammatory Response; Injections; Interferon Regulatory Factor 2; K-Series Research Career Programs; Knock-out; Lead; Link; Liver; Lymphocyte; Lymphocyte Activation; Lymphocyte Function; Lymphoid; MET gene; Malignant Neoplasms; Malignant neoplasm of liver; Modality; Modeling; Monitor; Mutate; Oncogenes; Patients; Phenotype; Physicians; Population; Predisposition; Primary carcinoma of the liver cells; Proto-Oncogenes; Receptor Protein-Tyrosine Kinases; Regimen; Regulation; Repression; Research Training; Residencies; Resistance; Role; Scientist; Signal Transduction; Sleeping Beauty; Survival Rate; System; Tail; Techniques; Testing; Therapeutic; Therapeutic Effect; Training; Transposase; Tumor Burden; Tumor Immunity; Tumor-infiltrating immune cells; United States; Unresectable; Veins; adaptive immune response; anti-PD1 therapy; beta catenin; biomarker driven; career; cell type; chronic liver disease; clinically relevant; combinatorial; differential expression; driver mutation; end stage disease; immunoregulation; improved; in vivo; inhibitor; liver cancer model; molecular subtypes; mortality; mouse model; mutant; mutant mouse model; neoplastic cell; novel; nuclear factor-erythroid 2; objective response rate; precision medicine; response; response biomarker; single-cell RNA sequencing; standard of care; synergism; synthetic biology; transcription factor; treatment stratification; tumor; tumor-immune system interactions; tumorigenesis

PROJECT SUMMARY/ABSTRACT Hepatocellular carcinoma (HCC) is the 6th leading cause of cancer-associated mortality in the United States, and is rising due to chronic liver disease and its associated sequalae. Currently, the response rates to current therapeutic paradigms consisting of immune checkpoint inhibitors (ICIs) remain low, and there exist an urgent need for novel combinatorial therapies to improve patient mortality. Lack of response to current ICIs is mainly due to a poor understanding of the tumor immune microenvironment (TIME) and how various HCC driver mutations lead to specific immune phenotypes. Additionally, there currently exist no biomarker-driven therapeutics for patient treatment stratification. ꞵ-catenin-active (encoded by mutated CTNNB1 oncogene) HCCs represent approximately 26-35% of HCCs and respond poorly to ICIs due to ꞵ-catenin driving an immunosuppressive TIME and limiting the effector function of lymphocytes important for anti-tumor immunity. We have developed novel ꞵ-catenin-mutated HCC mouse models where mutant CTNNB1 is co-expressed with either the proto-oncogene MET (ꞵ-catenin/hMet) or nuclear factor erythroid 2–related factor 2 (Nrf2) (ꞵ- catenin/Nrf2). These models represent 11% and 10% of all clinical HCC cases, respectively. Our preliminary studies demonstrate that ꞵ-catenin potentiates tumorigenesis in ꞵ-catenin-mutated HCC, and that directly targeting ꞵ-catenin promotes an inflammatory response driving anti-tumor immunity. Based on these observations, our overarching hypothesis is that ꞵ-catenin actively suppresses the adaptive immune response in the TIME and targeting ꞵ-catenin or its downstream immunomodulatory factors may improve susceptibility to ICIs. To investigate, I propose the following specific aims, which will uncover novel mechanisms of ꞵ-catenin signaling in the HCC TIME, aimed at developing precision medicine therapeutics. Specific Aim 1: We will determine the immune cells activated following ꞵ-catenin inhibition, and investigate whether there is in vivo synergy combining ꞵ-catenin inhibition and ICIs through single-cell RNA-sequencing (scRNA-seq) and multiplex immunohistochemistry. Thus, we will identify mechanisms of immunosuppression caused by ꞵ-catenin activation in ꞵ-catenin-mutated HCC. Specific Aim 2: Based on our preliminary data showing interferon regulatory factor- 2 (IRF2) repression in ꞵ-catenin-mutated HCC, we hypothesize that ꞵ-catenin-mutated HCCs may be sensitized to ICIs, or even show spontaneous tumor regression, upon re-expression of IRF2 as a result of enhanced immune response. We will use synthetic biology approaches to selectively induce IRF2 expression at various timepoints in tumorigenesis and monitor tumor burden. We will then use scRNA-seq on the lymphoid population to identify cell types and states regulated by IRF2, and test combination of IFNg (which induces IRF2) + ICI as a therapeutic modality. Contribution to Training: This proposal combines rigorous research training in liver cancer biology, immuno-oncology, and advanced bioinformatic analyses with an excellent clinical education, to aid my development as a future academic physician-scientist at the forefront of immune dysfunction in cancer.

项目摘要/摘要 在美国，肝细胞癌是癌症相关死亡的第六大原因， 由于慢性肝病及其相关后遗症，发病率正在上升。目前，对当前的响应率 由免疫检查点抑制剂(ICIS)组成的治疗方案仍然很低，存在一个紧迫的 需要新的组合疗法来提高患者死亡率。对当前ICIS缺乏反应主要是 由于对肿瘤免疫微环境(TIME)和各种肝细胞癌驱动因素了解不深 突变会导致特定的免疫表型。此外，目前还不存在生物标记物驱动的 治疗学对患者的治疗分层。ꞵ-连环蛋白活性(由突变的CTNNB1癌基因编码)肝癌 约占HCCs的26%-35%，由于ꞵ-catenin驱动 免疫抑制时间和限制淋巴细胞的效应功能对抗肿瘤免疫很重要。 我们已经建立了新的ꞵ-连环蛋白突变的肝癌小鼠模型，其中突变的CTNNB1与 原癌基因MET(ꞵ-连环蛋白/HMET)或核因子红系相关因子2(NRF2)(ꞵ-HMET)。 连环蛋白/NRF2)。这些模型分别占所有临床肝细胞癌病例的11%和10%。我们的预赛 研究表明，ꞵ-连环蛋白在ꞵ-连环蛋白突变的肝细胞癌中促进肿瘤发生，并且直接 以ꞵ-连环蛋白为靶点可促进炎症反应，从而促进抗肿瘤免疫。基于这些 观察到，我们的主要假设是ꞵ-catenin主动抑制获得性免疫反应。 在时间和靶向ꞵ-连环蛋白或其下游的免疫调节因子可能会提高易感性 ICIS。为了进行研究，我提出了以下具体目标，以揭示ꞵ-连锁素的新机制 在肝细胞癌时代发出信号，旨在发展精准医学疗法。具体目标1：我们将 检测ꞵ-连环蛋白抑制后激活的免疫细胞，并调查体内是否存在 单细胞ꞵ测序(scRNA-seq)和多重技术结合cRNA-catenin抑制和ICIS的协同作用 免疫组织化学。因此，我们将确定ꞵ-连环蛋白激活引起的免疫抑制的机制。 在ꞵ-连环蛋白突变的肝癌中。具体目标2：根据我们的初步数据显示干扰素调节因子- 2(Irf2)在ꞵ-连环蛋白突变的肝癌中的抑制，我们推测ꞵ-连环蛋白突变的肝癌可能是致敏的。 当IRF2作为增强的结果重新表达时，ICIS，甚至显示出自发的肿瘤消退 免疫反应。我们将使用合成生物学方法选择性地诱导IRF2在不同的 肿瘤发生的时间点和监测肿瘤负荷。然后我们将使用scRNA-seq来检测淋巴组织 鉴定受IRF2调控的细胞类型和状态，并测试IFNG(诱导IRF2)+ICI作为一种 治疗方式。对培训的贡献：这项建议结合了严格的肝脏研究培训 癌症生物学、免疫肿瘤学和先进的生物信息学分析以及出色的临床教育， 帮助我发展成为一名未来的学术内科医生兼科学家，站在癌症免疫功能障碍的前沿。