T-cell monitoring and immunotherapy for treating graft-versus-host disease

摘要 造血细胞移植（HCT）是一种潜在的治疗广泛的血液系统疾病的治疗方法。 然而，急性GVHD是限制其使用和结果的主要并发症 1 -3大约一半的急性GVHD患者会出现类固醇难治性， 一种死亡率特别高而治疗选择很少的疾病。因为目前没有 前瞻性地识别或治疗类固醇难治性GVHD的临床试验，治疗在很大程度上是徒劳的。 长期目标是联合收割机T细胞免疫监测和基于T细胞的 免疫疗法，以改善HCT患者的结果，并最终，许多其他患者。 在这个应用中，我们提出的工作可以重新定义我们如何理解，诊断和治疗GVHD， 对许多其他免疫疾病的影响。我们打算应用最先进的核苷酸测序技术- 基于方法来识别和跟踪T细胞引起GVHD。我们建议表明，这种知识可以 用于指导开发和实施改进的监测和新疗法，以治疗这些疾病， 患者这包括开发免疫抑制性T调节细胞来治疗GVHD 我们建议研究GVHD的临床前模型，并测试新的“受过教育的”供体CD 25 + T细胞是否具有以下特征： 似乎治疗活动性和严重的GVHD可能会转化为临床。这将是实现，如果 我们澄清了eT细胞的表型，并回答了这些细胞是否损害 临床相关环境中的移植物抗肿瘤反应和功能。我们预计，eTounder将需要 MHC相互作用的产生，较少依赖于IL-2发挥作用，并将防止GVHD的扩大。 通过T细胞库测序和/或对已知GVHD抗原的应答鉴定相关T细胞克隆。 我们还建议定义T细胞特征，预测，确认或分层患者的急性GVHD。我们 我建议将流式细胞术和单细胞基因组学整合到这种方法中，这些进展最终可以 允许临床医生在做出临床决策时加入适应性免疫系统的功能。我们将 如果我们证实TCR测序可用于对类固醇难治性进行风险分层， 患者或扩展以帮助预测GVHD。此外，如果我们能够确定 表型和治疗反应的T细胞高度参与GVHD，无论是对患者进行 常规HCT和接受THBE以预防或治疗GVHD的那些。的延伸 这项工作，我们已经设想了新的方法来确定GVHD靶抗原，通过整合高， 用于GVHD靶抗原发现的高通量TCR测序。 该提案将使科学家和临床从业者能够更好地理解和控制同种免疫， 希望有助于预防或治疗GVHD。

Abstract Hematopoietic cell transplantation (HCT) is a potentially curative therapy for a broad range of hematological malignancies and other disorders, however, acute GVHD is a major complication that limits the use and results in significant morbidity and mortality.1-3 About half of patients with acute GVHD will have steroid refractory disease that carries an especially high mortality with few treatment options. Because there are currently no clinical tests to prospectively identify or treat steroid refractory GVHD, therapy is largely empiric.2 My central long-term goal is to combine the use and development of T cell immune monitoring and T cell based immunotherapy to improve outcomes for HCT patients and, eventually, many other patients. In this application, we propose work that could redefine how we understand, diagnose and treat GVHD with implications for many other immunologic diseases. We propose to apply cutting-edge nucleotide sequencing- based approaches to identify and track T cells causing GVHD. We propose to show that this knowledge can be used to guide the development and implementation of improved monitoring and new therapies to treat these patients. This includes the development of immunosuppressive Tregulatory cells to treat GVHD We propose to study pre-clinical models of GVHD and to test if novel 'educated' donor CD25+ Tregs that appear to treat active and severe GVHD might be translated to the clinic. This will be accomplished if the phenotype of eTregs is clarified and we have answered the question as to whether or not these cells impair graft-versus tumor response and function in clinically relevant settings. We anticipate that eTregs will require MHC interactions to be generated, depend less upon IL-2 to function and will prevent the expansion of GVHD- associated T cell clones identified by T cell repertoire sequencing and/or response to known GVHD antigens. We propose also to defining T cell signatures that predict, confirm or stratify acute GVHD in patients. We propose to integrate flow cytometry and single cell genomics into this approach, advances that could finally allow clinicians to accession the function of the adaptive immune system in making clinical decisions. We will accomplish the goals of our study if we confirm TCR sequencing can be used to risk stratify steroid refractory patients or extended to help predict GVHD. In addition, the aim will be accomplished if we can identify the phenotype and treatment response of T cells highly implicated in GVHD, both for patients undergoing conventional HCT and for those receiving Tregs for the prevention or treatment of GVHD. As an extension of this work, we have conceived of novel approaches to identify GVHD target antigens by integrating high- throughput TCR sequencing for GVHD target antigen discovery. This proposal will allow scientists and clinical practitioners to better understand and control alloimmunity and hopefully help to prevent or cure GVHD.