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.

项目摘要 肝癌是世界上癌症相关死亡的第四大原因，死亡率正在上升， 这在一定程度上反映了历史上药物开发活动集中在单药治疗（多药治疗）上， 靶向激酶抑制剂和免疫疗法），其在患者中具有有限的功效。此外，还有一个 缺乏针对这种癌症类型的靶向个性化治疗选择。因此，迫切需要查明 新的定制治疗策略。肝癌靶向治疗的研究进展 (HCC)已经发现小分子抑制剂（例如，乐伐替尼）或 生物制剂（例如，贝伐单抗）与免疫疗法联合使用，尽管不到30%的 患者对这些新的一线疗法有反应。这一领域的一个主要差距是缺乏对相互间关系的认识， 患者异质性和未能识别（I）唯一针对驱动程序特定漏洞的药物，或（II） 改善免疫疗法。该提案的重点是肝癌治疗设计的未探索策略， 其利用了体内CRISPR筛选和一种新的患者来源的离体自体肿瘤的使用， 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，并允许评估联合治疗， 免疫疗法的改进。我假设药物在肝细胞癌中的进展，特别是在 与免疫疗法组合，依赖于药物介导的肿瘤T细胞调节 交互.这一假设将从两个方面进行检验：（1）剖析小说的作用机制 使用体内CRISPR筛选，在免疫学上易于处理的HCC模型中的候选药物WNTinib，和（2） 鉴定用于HCC个性化治疗的新型免疫疗法组合。对于目标1， WNTinib的关键靶点（或导致活性或耐药性的机制）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）转导遗传上确定的鼠HCC 原位移植到免疫活性和免疫受损小鼠中的类器官，和（II） 转导来自OT-I;Cas9和OT-II;Cas9小鼠的T细胞并将它们移植到表达 OVA衍生肽SIINFEKL。在WNTinib存在下进行的分析将识别WNTinib的肿瘤 细胞内在的、TME依赖的和T细胞依赖的靶或机制。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 肿瘤和T细胞作为单独成分和共同培养的研究。我将把这个平台扩展到 包括单独或与免疫疗法组合药物筛选，使用代表 HCC临床进展的各个阶段。该提案的结果将是向前迈出的一大步， 对HCC药物使用的多方面理解，以及药物如何与 免疫疗法提供个性化护理。