The regulation of tumour cell cytolysis by cancer associated fibroblasts

研究背景肿瘤微环境包括多种细胞类型，包括免疫细胞、成纤维细胞和内皮细胞。研究细胞毒性T淋巴细胞(CTL)和肿瘤相关成纤维细胞(CAF)之间的关系可以深入了解CTL和CAF在肿瘤进展中的相互作用和影响，也为治疗干预开辟了新的领域。CTL是一种免疫细胞，可以杀死癌细胞。然而，CTL的募集和激活在肿瘤中往往受到抑制。CAF塑造细胞外基质，并能分泌免疫调节和肿瘤促进/限制细胞因子。需要进一步的研究才能更好地了解CAF对CTL杀伤肿瘤细胞能力的影响。目标和目标该项目有两个主要目标。目标1专注于CAF，并询问不同的CAF如何与肿瘤细胞和免疫细胞，特别是CTL相互作用。我们将描述CAF的特征(特别是那些与乳腺癌和结直肠癌相关的CAF)，并在2D和3D共培养试验中研究它们支持癌细胞增殖的能力。同时，我们将评估CAF的免疫抑制功能，例如CAF分泌组对CTL增殖、迁移和CTL杀伤活性的影响。其目标是将CAF、肿瘤细胞和CTL结合到一个3D共培养模型中。目标2将重点研究新型双特异性T细胞结合蛋白(BITE)的活性。BITS将结合T细胞和肿瘤细胞上的受体，如EGFR或HER2。我们将比较内源性CTL和咬合定向CTL对肿瘤细胞的杀伤作用。该项目将进一步探讨不同水平的EGFR/HER2在癌细胞上的表达是否会影响BITE的活性。为了更好地了解CAF在治疗反应中的作用，我们将把目标1和目标2结合在一起，并在共培养系统中研究CAF对CTL和BIT导向的CTL诱导的肿瘤细胞溶解的影响。潜在的应用和益处CAF在许多实体肿瘤中都是隐含的，因此是癌症治疗的一个有吸引力的靶点。清楚地了解它们的功能以及与其他细胞的相互作用将有助于更好的药物设计。此外，研究双特异性分子在CAF/CTL环境中的作用机制将有助于未来免疫治疗的发展。与研究理事会相关这一博士项目在GW4-BioMed-2 DTP范围内。这是在“感染、免疫、抗菌素耐药性和修复”的主题范围内。拟议的项目与MRC投资于解决现实生活中生物复杂性的研究的战略是一致的，并与MRC的健康重点主题‘精确医学’和‘先进疗法’相关。此外，它还涉及GW4-BioMed-2 DTP概述的核心技能领域，包括量化和跨学科技能。

Context of researchThe tumour microenvironment includes a diverse number of cell types including immune cells, fibroblasts, and endothelial cells. Investigating the association between cytotoxic T lymphocytes (CTLs) and cancer associated fibroblasts (CAFs) could provide insights into the crosstalk and impact of CTLs and CAFs on tumour progression and also opens new areas for therapeutic intervention. CTLs are immune cells that can kill cancer cells. However, recruitment and activation of CTLs is often suppressed in the tumour. CAFs shape the extracellular matrix and can secrete immune-modulatory as well as tumour promoting/restricting cytokines. Further research is needed to better understand the impact of CAFs on CTLs ability to kill tumour cells. Aims and ObjectivesThe project has 2 major aims. Aim 1 focuses on CAFs and asks how different CAFs interact with tumour cells and immune cells, particularly CTLs. We will characterise CAFs (particularly those associated with breast and colorectal cancer) and investigate their ability to support cancer cell proliferation in 2D and 3D co-culture assays. In parallel, we will assess the immune-suppressive function of CAFs, e.g. the impact of the CAF secretome on CTL proliferation, migration and the cytolytic activity of CTLs. The objective is to then combine CAFs, tumour cells and CTLs into a 3D co-culture model. Aim 2 will focus on the activity of novel bispecific T-cell engagers (BiTEs). The BiTEs will bind both T-cells and receptors such as EGFR or HER2 on tumour cells. We will compare CTL-induced killing of tumour cells by endogenous CTLs and BiTE-directed CTLs. The project will further address whether expression of varying levels of EGFR/HER2 on cancer cells has an impact on the activity profile of the BiTEs. To better understand the role of CAFs on therapy response, we will bring aim 1 and 2 together, and investigate the impact of CAFs on CTL and BiTE-directed CTL induced tumour cell cytolysis in the co-culture system. potential application and benefitsCAFs are implicit in many solid tumours and are therefore an attractive target for cancer therapy. A clear understanding of their function and interaction with other cells would allow for better drug design. Furthermore, studying the mechanism of function of a bispecific molecule within the CAF/CTL environment will aid development of future immunotherapies. Relevance to the research councilThis is PhD project is within the GW4-BioMed-2 DTP. It is within the "Infection, immunity, antimicrobial resistance and repair" theme. The proposed project is aligned with the MRC's strategy to invest in research addressing biological complexity in real life and is relevant to the MRC's health focus themes of 'precision medicine' and 'advanced therapies'. Furthermore, it addresses core skill areas outlined by the GW4-BioMed-2 DTP including quantitative and interdisciplinary skills.