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

描述（由申请人提供）：造血细胞移植（HCT）是一种治疗多种血液恶性肿瘤和其他疾病的潜在疗法，然而，急性GVHD是一种主要并发症，限制了使用并导致显著的发病率和死亡率。1 -3约一半急性GVHD患者将患有类固醇难治性疾病，其死亡率特别高，治疗选择很少。由于目前还没有临床试验来前瞻性地识别或治疗类固醇难治性GVHD，因此治疗在很大程度上是徒劳的。2我的中心长期目标是将T细胞免疫监测和基于T细胞的免疫疗法的使用和开发结合起来，以改善HCT患者的结局，并最终改善许多其他患者的结局。在这个应用程序中，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与许多其他免疫疾病的影响。我们建议应用尖端的基于核苷酸测序的方法来鉴定和追踪引起GVHD的T细胞。我们建议表明，这些知识可用于指导改进监测和新疗法的开发和实施，以治疗这些患者。我们建议研究GVHD的临床前模型，并测试似乎治疗活动性和严重GVHD的新型“受过教育的”供体CD 25 + T细胞是否可以转化为临床。这将是完成，如果eT细胞的表型是明确的，我们已经回答了这个问题，这些细胞是否损害移植物抗肿瘤反应和功能，在临床相关的设置。我们预期eTclad将需要产生MHC相互作用，较少依赖于IL-2发挥功能，并且将阻止通过T细胞库测序和/或对已知GVHD抗原的应答鉴定的GVHD相关T细胞克隆的扩增。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，这些进步最终可以让临床医生在做出临床决策时加入适应性免疫系统的功能。如果我们确认TCR测序可用于对类固醇难治性患者进行风险分层或扩展以帮助预测GVHD，我们将实现我们的研究目标。此外，如果我们能够鉴定出与GVHD高度相关的T细胞的表型和治疗反应，则对于接受常规HCT的患者和接受THBG以预防或治疗GVHD的患者，都将实现该目标。作为这项工作的延伸，我们已经构思了通过整合用于GVHD靶抗原发现的高通量TCR测序来鉴定GVHD靶抗原的新方法。这一建议将使科学家和临床医生更好地了解和控制同种异体免疫，并希望有助于预防或治疗GVHD。

DESCRIPTION (provided by applicant): 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.