Biomarkers for assessing the immune state in rheumatoid arthritis and their application in a cellular therapy clinical trial

类风湿性关节炎(RA)是由自身免疫耐受丧失引起的。目前的临床生物标记物评估炎症，但随着我们更早地治疗以预防或治愈疾病，评估免疫状态将变得越来越重要。例如，我们开发了一种新的治疗方法，使用致耐受树突状细胞(TolDC)将致病的自身反应性T细胞转换为调节性/无能性T细胞。我们的I期临床试验显示这种疗法是安全的，我们即将进行的试验将比较不同的给药途径的疗效。为了证明TolDC的适当药效学作用，我们将开发评估免疫状态的分析方法，特别是自身反应性T细胞的功能能力。目的1)验证现有的和新的免疫监测工具用于表征来自RA患者的临床样本中的自身反应性T细胞的特征2)应用这些工具来探测与TolDC治疗有关的自身反应性T细胞的变化3)定义T细胞耐受性的生物标志物4)将我们的发现外推到其他临床RA情况新颖性在于结合最先进的技术来丰富自身反应性T细胞(包括使用人类白细胞抗原/多肽裂解物)并在单细胞水平上分析这些细胞，最终研究我们即将进行的tolDC临床试验中的T细胞，以展示治疗性免疫调节。时间一系列针对自身免疫性疾病的抗原特异性耐受疗法目前正在开发中，用于治疗和预防。这些策略包括基于细胞的(例如，tolDC)以及基于纳米颗粒的策略。然而，目前缺乏合适的药效学生物标记物，但这将是临床试验中展示免疫调节的基本结果指标。实验方法学生将验证已有的技术(多色流式细胞术、ELISPOT、体外增殖分析、单细胞测序)以及我们开发的丰富自身反应性T细胞的技术(人类白细胞抗原/多肽四聚体、激活诱导标记(AIM)分析)，用作临床样本的免疫监测工具。然后，这些分析将被应用于TolDC试验的样本，其中包括TolDC给药前后的外周血单核细胞以及淋巴结吸出物。作为一种意外情况，这些新工具还可以用于在其他相关场景中探索自身反应性T细胞的表型和功能，例如，当RA患者通过甲氨蝶呤等标准护理治疗进入缓解时，这些工具为疾病的免疫机制及其治疗的调节提供了宝贵的新见解。

BackgroundRheumatoid arthritis (RA) is caused by a loss of immune tolerance to self. Current clinical biomarkers assess inflammation but, as we treat earlier to prevent or cure disease, it will be increasingly important to assess the immune 'state'. For example, we have developed a novel therapeutic approach with tolerogenic dendritic cells (tolDC) that switch pathogenic self-reactive T-cells into regulatory/anergic T-cells. Our phase I clinical trial showed this therapy to be safe, and our forthcoming trial will compare different administration routes for efficacy. In order to demonstrate appropriate pharmacodynamic effects of tolDC, we will develop assays that assess the immune state, particularly the functional capabilities of autoreactive T-cells. Such surrogate outcome measures are the only way to provide true 'proof of concept' for such treatments.Objectives1) to validate existing and novel immunomonitoring tools for the characterisation of autoreactive T-cells in clinical samples from RA patients2) to apply these tools to probe changes in autoreactive-T-cells with tolDC treatment3) To define biomarkers for T-cell tolerance4) To extrapolate our findings to other clinical RA scenariosNoveltyThe novelty lies in combining state-of-the-art techniques to enrich autoreactive T-cells (including the use of HLA/peptide tetramers) and analysing those cells at single cell level, ultimately studying T-cells from our forthcoming tolDC clinical trial to demonstrate therapeutic immune modulation. TimelinessA number of antigen-specific tolerogenic therapies for autoimmune diseases are currently under development, for treatment and prevention. These include cell-based (e.g. tolDC) as well as nanoparticle-based strategies. However, suitable pharmacodynamic biomarkers are currently lacking, but will be essential outcome measures in clinical trials to demonstrate immune modulation.Experimental approachThe student will validate established techniques (multi-colour flow cytometry, ELISPOT, in vitro proliferation assay, single cell sequencing) alongside techniques we have developed to enrich autoreactive T-cells (HLA/peptide tetramers, activation-induced marker (AIM) assay) for use as immunomonitoring tools for clinical samples. These assays will then be applied to samples from the tolDC trial which include peripheral blood mononuclear cells, as well as lymph node aspirates, taken before and after tolDC administration. As a contingency, these novel tools can also be applied to probe the phenotype and function of autoreactive T-cells in other relevant scenarios, for example, from RA patients as they enter remission with standard-of-care treatment such as methotrexate, providing invaluable new insights into immune mechanisms of disease and their modulation by treatment.