EPSRC Centre for Doctoral Training in Engineered Tissues for Discovery, Industry and Medicine

免疫检查点阻断剂（ICBs）调节针对癌症的免疫反应，彻底改变了癌症治疗，但当肿瘤转移到肝脏时，ICBs的疗效很低1。这是一个巨大的挑战，因为肝脏是继发性肿瘤最常见的部位之一。此外，只有20%的原发性肝癌患者对免疫治疗有反应2。因此，肝脏微环境似乎是抑制肿瘤特异性反应的主要驱动因素，使癌症治疗效率降低。肝脏是一个致耐受性器官，肝窦内皮细胞（LSEC）排列在肝脏的细血管上，对致耐受性微环境做出关键贡献3。LSEC可以改变免疫细胞功能，并作为免疫细胞浸润到肝脏中的看门人，控制哪些免疫细胞亚群从循环中招募4。LSEC的重要性在组织再生和癌症过程中经常被忽视，我们有初步数据显示，人类肝内皮细胞被肿瘤微环境重新编程，并可能形成效应T细胞募集/激活的屏障。人类肝脏肿瘤微环境的研究具有挑战性，因为动物模型不能完全概括人类疾病的原因和机制。例如，免疫成分和对疾病的反应的差异。为了克服这一局限性，我们的目标是港口再生医学中使用的领先技术，通过开发一个三维的人类肝脏肿瘤模型，以调查肿瘤上皮细胞，内皮细胞和免疫celles.Aim之间的串扰1开发一个培养系统与细胞成分来自相同的患者，重演体内肿瘤微环境。另外还建立了诸如Transwell和Ibidi贩运分庭等替代性共文化制度。 目的2探讨肿瘤微环境对T细胞运输的影响。T细胞将从患者的血液样品中分离，并与类器官一起培养。将在与源自患者或非肿瘤供体的类器官一起培养后研究T细胞运输和表型。目标3我们将测试在这个3D肿瘤/内皮模型中，抑制先前目标中确定的内皮因子是否会导致与免疫检查点抑制剂对T细胞浸润的协同作用。学生将学习一系列技术，并从组织收集，细胞分离和模型生成中获得实践经验。这些包括组织学分析、通过FACS的细胞分离、细胞/类器官培养、分子和转录分析。培训机会将通过在体外环境中使用最先进的跨学科技术来支持癌症治疗的新方法，以1）开发患者衍生的新型3D肝脏模型2）识别肿瘤影响内皮细胞/ T细胞串扰的分子途径。学生将通过主管Shetty博士与NHS和国家健康研究所密切联系。Shetty博士是伯明翰伊丽莎白女王医院的一名肝病专家顾问，专门研究肝癌。Shetty博士参与该项目旨在解决目前临床上缺乏的问题，只有20%的肝癌患者对免疫检查点阻断治疗有反应。这将产生临床相关和可翻译的输出，未来可能对人类健康产生积极影响。在学生发展方面，学生将在基础和转化研究方面拥有丰富的经验，将生物医学科学的基础知识与化学工程相结合。学生将受益于研究领域的广度和多学科性，并受到该项目目标的激励，该项目旨在改善患者健康。

Immune checkpoint blockers (ICBs) which regulates immune response against cancer have revolutionized cancer therapy, but the efficacy of ICBs is low when tumours metastasise to the liver1. This is a huge challenge since the liver is one of the commonest sites for secondary tumours. Furthermore, only 20% of primary liver cancer patients are responsive to immunotherapy2. The liver microenvironment therefore appears to be a major driver in suppressing tumour specific responses making cancer therapy less efficient.The liver is a tolerogenic organ with liver sinusoidal endothelial cells (LSEC), which line the fine blood vessels in the liver, making a critical contribution to the tolerogenic microenvironment3. LSEC can alter immune cell function and act as the gatekeeper to immune cell infiltration into the liver, controlling which immune cell subsets are recruited from the circulation4. The importance of LSEC is often overlooked during tissue regeneration and cancer, and we have pilot data showing that human liver endothelium is reprogrammed by the tumour microenvironment and likely to form a barrier to effector T cell recruitment/activation. The investigation of human liver tumour microenvironment is challenging as animal models do not fully recapitulate the cause and mechanisms of human disease. For example, the differences in immune components and response against disease. To overcome this limitation, we aim to harbour the leading technologies used in regenerative medicine by developing a three-dimensional human liver tumour model to investigate the crosstalk between the tumour epithelium, endothelial cells, and the immune cells.Aim 1 To develop a culture system with cellular components derived from the same patients to recapitulate tumour microenvironment in vivo. Alternative co-culture system such as the Transwell and the Ibidi trafficking chambers are also in place. Aim 2 To investigate the effect of tumour microenvironment on T-cell trafficking. T-cells will be isolated from blood samples obtained from patients and culture with the organoids. T-cell trafficking and phenotype will be investigated after culturing with organoids derived from patients or non-tumourous donor. Aim 3 We will test whether inhibiting the endothelial factors identified in previous aims leads to a synergistic effect with immune checkpoint inhibitors on T cell infiltration in this 3D tumour/endothelial model.The student will learn a range of techniques with hands on experience from tissue collection, cell isolation and model generation. These include, histological analysis, cell isolation by FACS, cell/organoid culture, molecular and transcriptional analysis. The training opportunities will support novel approaches to cancer therapy by using state of the art interdisciplinary techniques in an in vitro setting to 1) develop a patient-derived novel 3D liver model 2) Identify molecular pathways by which tumours affect endothelial cell/ T cell crosstalk.The student will have close link to the NHS and National Institute for Health Research through the supervisor Dr Shetty. Dr Shetty is a consultant hepatologist who specialized in liver cancer at the Queen Elizabeth Hospital, Birmingham. The involvement of Dr Shetty in this project aims to tackle what is currently lacking in clinics, with only 20% of liver cancer patients responds to immune checkpoint blockade treatment. This will generate outputs which is clinically relevant and translatable which can have a positive impact on human health in the future. In terms of student development, the student will have a wealth of experience in basic and translational research, incorporating the fundamentals of biomedical sciences with chemical engineering. The student will benefit from the breadth and multidisciplinary of research areas, and motivated by the goals of this project which directs to improve patient health.