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

肿瘤消融手术，如微波消融和冷冻消融，为肝细胞癌（HCC）提供了局部肿瘤控制，但受到周围肝硬化背景肝脏损伤的限制。组织切片术是一种新的非侵入性和高度靶向的肿瘤消融术，它可以改善局部肿瘤控制，同时使肝功能虚弱的患者从肿瘤消融术中受益。此外，了解肝细胞癌对组织学检查的免疫反应将有助于开发靶向免疫治疗方案，以防止后续肿瘤的出现，同时保护脆弱的背景肝脏。比较微波和冷冻消融后肿瘤免疫组织学特征。设计新的肝细胞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：本项目将包括以下实验主题。肝脏手术患者原发性肝癌和非恶性肝样本的组织培养。2 .肝癌患者微波/冷冻消融或组织切片后免疫检查点分子在辅助/细胞毒性t细胞、NK细胞和抗原提呈细胞上的时间表达动态的流式细胞术评估和血浆细胞因子定量。通过RNAseq， TCRseq和功能性免疫分析评估组织切片后先天免疫和适应性免疫抗肿瘤作用的机制。在体外t细胞启动模型中优化组织免疫治疗组合和时间表，以测试非恶性肝细胞的抗癌功效和背景肝安全性。免疫治疗联合肿瘤消融治疗双侧侧腹同种肝癌模型的体内模型。肝细胞癌组织切片法联合免疫治疗早期临床试验的合理设计。影响：Hope4Liver研究是英国首次应用组织切片法。与传统的微波和冷冻消融技术相比，该项目首次描述了组织切片术的免疫后遗症。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，为HCC患者和更广泛的肿瘤类型定义和完善未来的肿瘤消融免疫治疗方案。

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.