An immunocompetent platform for the characterization and discovery of novel therapeutics for hepatocellular carcinoma

用于表征和发现肝细胞癌新疗法的免疫活性平台

• 批准号: 10507168
• 负责人: Alexander Rialdi
• 金额: $ 17.15万
• 依托单位: ICAHN SCHOOL OF MEDICINE AT MOUNT SINAI
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-01 至 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10507168
• 关键词: Address; Animal Model; Antigens; Autologous; BAY 54-9085; Back; Biological; Biological Products; Biopsy; CRISPR screen; CRISPR/Cas technology; CTNNB1 gene; Cancer Biology; Cancer Etiology; Cell Communication; Cell Culture Techniques; Cell model; Cessation of life; Chemicals; Clinical; Clustered Regularly Interspaced Short Palindromic Repeats; Coculture Techniques; Combination immunotherapy; Combined Modality Therapy; Cues; Data; Dissection; Drug Combinations; Drug Screening; Drug Targeting; Drug usage; EZH2 gene; Ensure; Epigenetic Process; FDA approved; Failure; Funding; Goals; Heterogeneity; Immunocompetent; Immunocompromised Host; Immunologics; Immunotherapy; In Vitro; Investigation; Knock-out; Laboratories; Libraries; Malignant neoplasm of liver; Mediating; Mentors; Modality; Modeling; Mus; Mutate; Mutation; Nature; OVA-8; Organoids; Ovalbumin; Patients; Pharmaceutical Preparations; Phase; Physicians; Physiological; Primary carcinoma of the liver cells; Protocols documentation; Refractory; Reporting; Research; Research Personnel; Resistance; Signal Transduction; T-Lymphocyte; Testing; Therapeutic; Transplantation; Tumor-Derived; Tumor-Infiltrating Lymphocytes; Validation; Work; anti-PD-1; beta catenin; bevacizumab; cancer type; chemotherapy; clinically translatable; drug candidate; drug development; drug use screening; gene discovery; genome-wide; immune checkpoint blockade; improved; in vivo; individual patient; individualized medicine; insight; kinase inhibitor; liver cancer model; mortality; mouse genome; neoplastic cell; novel; novel therapeutics; organoid transplantation; personalized care; personalized medicine; personalized therapeutic; resistance mechanism; screening; skills; small molecule inhibitor; success; therapeutic target; therapy design; tool; treatment strategy; tumor; tumor immunology; tumor microenvironment

Project Summary Liver cancer is the fourth leading cause of cancer-related death in the world and mortality rates are rising, which, in part, reflects historical drug development campaigns focused on single agent therapies (multi- targeted kinase inhibitors and immunotherapy) that have limited efficacy in patients. Additionally, there are a lack of targeted, personalized therapeutic options for this cancer type. Thus, there is an urgent need to identify novel tailored therapeutic strategies. Recent advances in the therapeutic targeting of hepatocellular carcinoma (HCC) have discovered superiority in efficacy for either small molecule inhibitors (for example, lenvatinib) or biologics (for example, bevacizumab) used in combination with immunotherapy, though less than 30% of patients respond to these new frontline therapies. A major gap in the field is the lack of appreciation for inter- patient heterogeneity and a failure to identify drugs that (I) uniquely target driver-specific vulnerabilities, or (II) improve immunotherapy. This proposal focuses on an unexplored strategy for therapy design in liver cancer, which leverages the usage of in vivo CRISPR screens and a novel patient-derived ex vivo autologous tumor organoid-T cell co-culture platform to provide new biological insight to mechanisms of drug potency on two critical components of the tumor microenvironment (TME), and allow assessment of combination therapies for the improvement of immunotherapy. I hypothesize that drug advancement in HCC, particularly in combination with immunotherapy, is dependent on drug-mediated modulation of tumor-T cell interactions. This hypothesis will be tested in two aims: (1) dissection of the mechanism of action of a novel candidate drug, WNTinib, in immunologically tractable models of HCC using in vivo CRISPR screens, and (2) identification of novel immunotherapy combinations for the personalized treatment of HCC. For Aim 1, WNTinib’s key targets (or mechanism’s contributing to activity or resistance) will be investigated using two separate genome-scale CRISPR sgRNA screening strategies: (I) transducing genetically-defined murine HCC organoids transplanted orthotopically into immunocompetent and immunocompromised mice, and (II) transducing T cells from OT-I;Cas9 and OT-II;Cas9 mice and transplanting them into tumor models expressing the OVA-derived peptide SIINFEKL. Analyses done in the presence of WNTinib will identify WNTinib’s tumor cell intrinsic, TME-dependent, and T cell-dependent targets or mechanisms. For Aim 2, I will use HCC patient tumor biopsies to establish organoids and tumor infiltrating lymphocyte (TIL) cultures. This approach will allow the study of tumor and T cells as separate components and in co-culture together. I will expand this platform to include drug screening, alone or in combination with immunotherapy, using a panel of drugs representing various stages of HCC clinical advancement. Results from this proposal will represent a major step forward for the multidimensional understanding of drug usage in HCC, and how drugs may be used in combination with immunotherapy to provide personalized care.

项目概要 肝癌是世界上第四大癌症相关死亡原因，死亡率正在上升， 这在一定程度上反映了历史上专注于单药治疗（多药治疗）的药物开发活动 靶向激酶抑制剂和免疫疗法）对患者的疗效有限。此外，还有一个 对于这种癌症类型缺乏有针对性的、个性化的治疗选择。因此，迫切需要识别 新颖的定制治疗策略。肝细胞癌靶向治疗的最新进展 （HCC）发现小分子抑制剂（例如乐伐替尼）或 生物制剂（例如贝伐珠单抗）与免疫疗法联合使用，但只有不到 30% 患者对这些新的一线疗法有反应。该领域的一个主要差距是缺乏对相互之间的认识。 患者异质性和未能识别 (I) 专门针对驾驶员特定漏洞的药物，或 (II) 改善免疫治疗。该提案重点关注肝癌治疗设计的未探索策略， 它利用体内 CRISPR 筛选和新型患者体外自体肿瘤 类器官-T 细胞共培养平台为两种药物的药效机制提供新的生物学见解 肿瘤微环境（TME）的关键组成部分，并允许评估联合疗法 免疫治疗的改进。我假设 HCC 药物的进步，特别是在 与免疫疗法相结合，依赖于药物介导的肿瘤 T 细胞调节 互动。该假设将在两个目标上进行检验：（1）剖析小说的作用机制 使用体内 CRISPR 筛选在 HCC 免疫学易处理模型中使用候选药物 WNTinib，以及 (2) 鉴定用于 HCC 个性化治疗的新型免疫治疗组合。对于目标 1， WNTinib 的关键目标（或对活性或耐药性的贡献机制）将使用两种方法进行研究 单独的基因组规模 CRISPR sgRNA 筛选策略：(I) 转导基因定义的小鼠 HCC 将类器官原位移植到免疫功能正常和免疫功能低下的小鼠中，以及（II） 转导来自 OT-I;Cas9 和 OT-II;Cas9 小鼠的 T 细胞并将其移植到表达表达的肿瘤模型中 OVA 衍生肽 SIINFEKL。在 WNTinib 存在的情况下进行的分析将识别 WNTinib 的肿瘤 细胞内在的、TME 依赖性的和 T 细胞依赖性的靶标或机制。对于目标 2，我将使用 HCC 患者 肿瘤活检以建立类器官和肿瘤浸润淋巴细胞（TIL）培养物。这种方法将允许 将肿瘤和 T 细胞作为单独的成分和一起共培养的研究。我会把这个平台拓展到 包括单独或与免疫疗法组合的药物筛选，使用一组代表的药物 HCC 临床进展的各个阶段。该提案的结果将代表着向前迈出的重要一步 对 HCC 药物使用的多维理解，以及药物如何与 HCC 联合使用 免疫疗法提供个性化护理。