Targeting the mutanome of HCC by viral inflammation and tumor-directed vaccinations: a model for individualized tumor therapy

通过病毒炎症和肿瘤定向疫苗接种来靶向 HCC 突变组：个体化肿瘤治疗模型

• 批准号: 279621100
• 负责人: Professor Dr. Florian Kühnel
• 金额: --
• 依托单位: Klinik für Gastroenterologie, Hepatologie und Endokrinologie
• 依托单位国家: 德国
• 项目类别: Research Grants
• 财政年份: 2015
• 资助国家: 德国
• 起止时间: 2014-12-31 至 2018-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/279621100?language=en
• 关键词: Targeting; mutanome; HCC; viral; inflammation

Oncolytic virotherapy and immunotherapies promise enormous potential for the treatment of hepatocellular carcinoma (HCC). In previous experiments we have developed an innovative and highly effective immunotherapeutic strategy which facilitates an effective tumor-targeted dendritic cell vaccination during inflammation of the tumor by an oncolytic virus (= oncolysis-assisted DC-vaccination, or ODC). In further preliminary work in a lung cancer cell we established a diagnostic/therapeutic procedure involving identification of the tumor mutanome by whole exome sequencing, followed by in-silico prediction of potentially immunogenic neoepitopes and verification by T cell analyses after intratumoral infection with oncolytic adenoviruses. In the proposed project we want to develop a personalized therapy strategy for the treatment of HCC involving an optimized ODC strategy to elicit strong antitumoral immune response against immunogenic neoepitopes of individual tumor mutanomes. We want to address the therapeutic efficacy of optimized ODC against immunogenic neoepitopes in clinically relevant, transgenic mouse models. First, we want to analyse the spectrum of non-synonymous single nucleotide variants in selected HCC cell lines by whole exome sequencing. After determination of the mutanome, we define promising MHC class I-binding neoepitopes using the SYFPEITHI algorithm. Successful triggering of CD8 T cell responses against immunogenic neoepitopes will be identified by CD8 T cell screens after oncolytic virus infection of subcutaneous tumors in a syngeneic mouse model. In these models, we want to use these immunogenic neoepitopes to define the rules for optimized preparation of DC-vaccines targeting a complete mutanome. Antigen-specific immune responses will be determined by ELISpot analyses and in-vivo cytotoxicity measurements. Therapeutic efficacy will be assessed by monitoring tumor growth, survival, and metastases formation. Finally, we want to investigate the therapeutic potential of corresponding ODC-based vaccine in our novel electroporation model for transgenic intrahepatic HCC. These investigations will provide essential informations for a future clinical trial of personalized immunotherapy of HCC.

溶瘤病毒治疗和免疫治疗在治疗肝细胞癌方面具有巨大的潜力。在以前的实验中，我们开发了一种创新和高效的免疫治疗策略，该策略有助于在肿瘤溶瘤病毒(ODC)炎症期间有效地进行肿瘤靶向树突状细胞疫苗接种(=溶瘤辅助DC疫苗，或ODC)。在肺癌细胞的进一步前期工作中，我们建立了一种诊断/治疗程序，包括通过全外显子组测序鉴定肿瘤突变组，然后对潜在的免疫原性新表位进行电子预测，并在肿瘤内感染溶瘤腺病毒后通过T细胞分析进行验证。在拟议的项目中，我们希望开发一种个性化的治疗肝癌的策略，包括一种优化的ODC策略，以诱导针对单个肿瘤突变体免疫原性肿瘤表位的强大抗肿瘤免疫反应。我们希望在临床相关的转基因小鼠模型中解决优化的ODC对免疫原性新表位的治疗效果。首先，我们想要通过整个外显子组测序来分析选定的肝癌细胞系中非同义的单核苷酸变异的谱。在确定突变组后，我们使用SYFPEITHI算法定义了有前景的MHC I类结合新表位。在同基因小鼠模型中，溶瘤病毒感染皮下肿瘤后，CD8T细胞筛选将鉴定成功触发针对免疫原性新表位的CD8T细胞反应。在这些模型中，我们希望使用这些免疫原性新表位来定义针对完整突变组的DC疫苗的优化制备规则。抗原特异性免疫反应将通过ELISpot分析和体内细胞毒性测量来确定。治疗效果将通过监测肿瘤的生长、存活率和转移形成来评估。最后，我们想要研究相应的基于ODC的疫苗在我们新的转基因肝内肝癌电穿孔模型中的治疗潜力。这些研究将为未来肝癌个体化免疫治疗的临床试验提供必要的信息。