Exploiting Tumour Ablation Histotripsy to Augment anti-cancer Immunotherapy in Hepatocellular Carcinoma

肿瘤消融术，如微波消融术和冷冻消融术，为肝细胞癌(HCC)提供了局部肿瘤控制，但受到周围肝硬变背景肝损害的限制。组织摩擦学是一种新的非侵入性和高度靶向性的肿瘤消融术，由组织电子公司正在开发，它将改善局部肿瘤控制，同时允许肝功能虚弱的患者从肿瘤消融术中受益。此外，了解肝细胞癌组织摩擦后的免疫反应将有助于开发有针对性的免疫治疗方案，以防止后续肿瘤的出现，同时保护脆弱的背景肝脏。目的：1.与微波和冷冻消融相比，组织摩擦后的肿瘤免疫学特征。设计新的肝癌消融后免疫治疗方案，以诱导强大和持久的抗癌免疫，同时限制对脆弱的背景肝脏的副作用。实验方法：该项目将包括以下实验主题。方法1.肝手术患者的原发性肝细胞癌和非恶性肝组织标本的组织培养。流式细胞仪检测和血浆细胞因子定量检测肝细胞癌患者微波/冷冻或组织学治疗前后辅助性/细胞毒性T细胞、NK细胞和抗原提呈细胞表面免疫检查点分子表达的动态变化。用RNAseq、TCRseq和功能免疫分析评价组织学后先天免疫和获得性免疫抗肿瘤作用的机制。在体外T细胞启动模型中组织摩擦学-免疫治疗组合和时间表的优化，以测试非恶性肝细胞的抗癌有效性和背景肝脏安全性。在双侧同种异体肝细胞癌模型中联合免疫治疗和肿瘤消融的体内模型。合理设计免疫治疗与组织学联合应用于肝癌的早期临床试验影响：Hope4Liver研究是组织学在英国的第一次应用。与传统的微波和冷冻消融技术相比，这个项目是第一次描述组织摩擦的免疫后遗症。该项目的成功完成将直接定义和完善肝细胞癌免疫治疗-组织学联合研究。因此，这将是一个改变实践的博士项目，定义和提炼未来的肿瘤消融-免疫治疗方案，用于肝癌患者和更广泛的肿瘤类型。

Tumour ablative procedures, such as microwave ablation and cryoablation, offer localised tumour control for hepatocellular carcinoma (HCC) but are limited by damage to the surrounding cirrhotic background liver. Histotripsy is a new non-invasive and highly targeted tumour ablation procedure under development by Histosonics, which stands to improve local tumour control whilst allowing patients with frail liver function to benefit from tumour ablation. Furthermore, understanding the immune response to histotripsy in HCC would enable targeted immunotherapy regimens to be developed to prevent the emergence of subsequent tumours, whilst protecting the frail background liver.Objectives:1. To characterise tumour immunology post-histotripsy in comparison to microwave- and cryoablation.2. To design novel immunotherapy regimens post-ablation in HCC to induce robust and long-lasting anti-cancer immunity, whilst limiting side-effects to the frail background liver.Experimental Approach:The project will include the following experimental themes.1. Tissue culture of primary HCC and non-malignant liver samples derived from patients undergoing hepatic surgery.2. Flow cytometry assessment and plasma cytokine quantification of the temporal expression dynamics of immune checkpoint molecules on helper/cytotoxic T-cells, NK cells and antigen presenting cells, post-microwave-/cryo-ablation or histotripsy in HCC patients.3. Assessment of the mechanisms of innate and adaptive immune anti-tumour effects post-histotripsy, by RNAseq, TCRseq and functional immune assays.4. Optimisation of histotripsy-immunotherapy combinations and schedules in an in vitro T-cell priming model to test anti- cancer efficacy and background liver safety in non-malignant hepatocytes.5. In vivo models of combination immunotherapy with tumour ablation in a bilateral flank syngeneic model of HCC.6. Rational design of an early-phase clinical trial of combination immunotherapy with histotripsy in HCC.Impact:The Hope4Liver study is the first application of histotripsy in the UK. This project is the first to characterise the immunological sequelae of histotripsy, in comparison to conventional microwave- and cryoablation techniques. Successful completion of this project will directly define and refine a combination immunotherapy-histotripsy study in HCC. This therefore stands to be a practice-changing PhD project, defining and refining future tumour ablation-immunotherapy treatment regimens for patients with HCC and wider tumour types.