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

摘要 造血细胞移植(Hct)是一种治疗多种血液病的潜在疗法。 然而，恶性肿瘤和其他疾病，急性移植物抗宿主病是限制使用和结果的主要并发症。 1-3大约一半的急性移植物抗宿主病患者会有类固醇耐药 死亡率特别高，几乎没有治疗选择的疾病。因为目前没有 临床试验前瞻性地识别或治疗类固醇难治性移植物抗宿主病，治疗在很大程度上是经验性的。 远期目标是将T细胞免疫监测和T细胞为基础的使用和发展结合起来 免疫治疗以改善HCT患者的预后，并最终改善许多其他患者的预后。 在这项申请中，我们提出了一项工作，可以重新定义我们如何理解、诊断和治疗GVHD 对许多其他免疫性疾病的影响。我们建议应用尖端的核苷酸测序- 基于识别和跟踪引起移植物抗宿主病的T细胞的方法。我们打算证明，这种知识可以 用于指导开发和实施改进的监测和治疗这些疾病的新疗法 病人。这包括开发免疫抑制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捐赠者CD25+Tregs是否 看来治疗活动期和重度GVHD可能会转化为临床。这将在以下情况下实现 ETregs的表型已经明确，我们已经回答了这些细胞是否会损伤的问题 临床相关环境中移植物抗肿瘤的反应和功能。我们预计eTregs将需要 MHC相互作用的产生，较少依赖IL-2发挥作用，并将防止GVHD的扩张- 通过T细胞谱系测序和/或对已知GVHD抗原的反应识别相关的T细胞克隆。 我们还建议定义T细胞信号来预测、确认或分层患者的急性移植物抗宿主病。我们 建议将流式细胞术和单细胞基因组学整合到这一方法中，最终可能取得的进展 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。我们会 如果我们确认TCR测序可以用于对类固醇难治性进行风险分层，就可以实现我们的研究目标 患者或扩展以帮助预测移植物抗宿主病。此外，如果我们能够识别出 T细胞的表型和治疗反应与移植物抗宿主病密切相关，两者都适用于 适用于常规HCT和接受Tregs预防或治疗GVHD的患者。作为对 在这项工作中，我们设想了一种新的方法来识别GVHD靶抗原，方法是将高表达的 通过TCR测序发现GVHD靶抗原。 这一建议将使科学家和临床从业者更好地了解和控制同种异体免疫和 希望有助于预防或治愈移植物抗宿主病。

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.