Novel GvHD-Specific Gene Identification for Optimal Control of GvHD

摘要 移植物抗宿主病（GvHD）是同种异体造血干细胞移植成功的最重要障碍之一。 细胞移植（allo-HCT）。该项目的目标是确定新的基因，其遗传/药理学 阻断可以选择性地预防GvHD，同时维持或增强抗白血病活性（aka graft- 抗白血病或GvL效应）。Allo-HCT仍然是血液病患者最有效的治疗方法。 在恶性肿瘤中，因为allo-HCT的治疗益处主要来源于GvL。然而，allo-HCT也 存在移植物中的供体T细胞变得过度活跃并开始攻击非供体T细胞的风险。 不仅白血病，而且患者的皮肤，肠道和肝脏，导致GvHD。因为一个强大的 两种供体T细胞介导的免疫应答之间的关联，特别是限制GvHD仍然是一个问题。 allo-HCT的目标。管理GvHD的威胁将扩大allo-HCT的范围和益处。几 包括我们在内的研究小组已经提出了治疗策略，以减少GvHD，而不废除有益的 动物模型和人类患者中的GvL效应。然而，同种异体供体T细胞 差异调节GvHD和GvL在很大程度上仍然未知。我们对机械论的理解 阻碍了我们特异性预防/治疗GvHD的能力。我们假设我们认为是 与浸润肿瘤的供体T细胞相对于GvHD器官的供体T细胞的差异相关将是免疫治疗的关键靶点。 预防GvHD而不负面影响GvL。为此，我们进行了一个无偏的全基因组 CRISPR/Cas9文库筛选。因此，我们首次鉴定了487个新的GvHD相关基因。 这些基因的指导RNA被富集， 仅在得自肿瘤结节而非得自任何GvHD靶器官的T细胞中。我们发现五组 参与已知途径的基因在这487个基因中最显著地富集。该五项 通路与在同种异体移植中尚未积极研究的JN-RAP 1-FGFR 3轴密切相关， HCT字段。因此，我们将确定是否遗传/药理学抑制的JN-RAP 1-FGFR 3轴导致， GvHD特异性免疫调节，同时维持或增强我们的小鼠模型中的GvL。 GvHD/GvL和人PBMC/T细胞介导的异种-GvHD/GvL。如果成功，我们对新型治疗方法的发现 选择性和最佳控制GvHD的目标将是潜在的游戏规则改变者。我们建议的研究将 为GvHD及其预防提供了重要的机制见解，这可能具有变革性意义。 不仅对实体器官移植排斥反应而且对炎症性排斥反应的治疗发展的意义 炎症性肠病、牛皮癣和类风湿性关节炎等疾病。此外，我们的研究可能 也有助于提高抗癌疗法和免疫疗法的功效， .

ABSTRACT Graft-versus-host disease (GvHD) is one of the most significant barriers to success for allogeneic hematopoietic cell transplantation (allo-HCT). The goal of this project is to identify novel genes whose genetic/pharmacologic blockade may selectively prevent GvHD while maintaining or enhancing anti-leukemia activities (aka graft- versus-leukemia or GvL effect). Allo-HCT remains the most effective treatment for patients with hematologic malignancies, as the therapeutic benefit of allo-HCT is primarily derived from GvL. However, allo-HCT also carries the risk that the donor T cells in the transplant (graft) will become overzealous and begin to attack not only the leukemia but also the patient's skin, intestines and liver, resulting in GvHD. Because of a strong association between the two donor T cell-mediated immune responses, specifically limiting GvHD remains the goal of allo-HCT. Managing the threat of GvHD would broaden the scope and benefit of allo-HCT. Several groups, including ours, have proposed therapeutic strategies to reduce GvHD without abrogating the beneficial GvL effect in animal models and human patients. However, the mechanisms by which allogeneic donor T cells differentially modulate GvHD and GvL remain largely unknown. This gap in our mechanistic understanding hinders our ability to specifically prevent/treat GvHD. We hypothesize that the genes we identify as being differentially associated with donor T cells that infiltrate tumors versus GvHD organs will be critical targets for the prevention of GvHD without negatively affecting GvL. To this end, we performed an unbiased genome-wide CRISPR/Cas9 library screen. As a result, we have for the first time identified 487 novel GvHD-associated genes that could potentially serve as molecular targets in GvHD, as the guide RNAs for these genes were enriched only in T cells obtained from tumor nodules but not from any GvHD target organs. We found that five sets of genes that are involved in known pathways are most significantly enriched in these 487 genes. All of these five pathways are closely linked to the RELN-RAP1-FGFR3 axis that has not been actively investigated in the allo- HCT field. Thus, we will determine if genetic/pharmacologic inhibition of the RELN-RAP1-FGFR3 axis results in GvHD-specific immune modulation while maintaining or enhancing GvL in our mouse models of murine GvHD/GvL and human PBMC/T cell-mediated xeno-GvHD/GvL. If successful, our findings of novel therapeutic targets to selectively and optimally control GvHD will be potential game-changers. Our proposed studies will provide significant mechanistic insights into GvHD and its prevention, which may have transformative implications for therapeutic development not only for solid organ transplant rejection but also inflammatory diseases such as inflammatory bowel disease, psoriasis and rheumatoid arthritis. In addition, our studies may also help improve the efficacies of anti-cancer therapies and immunotherapeutic approaches .