JAK2_STAT3 as a therapeutic target and marker of graft-versus-host disease

JAK2_STAT3 作为移植物抗宿主病的治疗靶点和标志物

• 批准号: 8829894
• 负责人: Brian C Betts
• 金额: $ 12.58万
• 依托单位: H. LEE MOFFITT CANCER CTR & RES INST
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-08-10 至 2016-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8829894
• 关键词: Acute Graft Versus Host Disease; Allogenic; Biological; Biological Markers; Biological Preservation; Bone Marrow; Bone Marrow Transplantation; CD8B1 gene; Cells; Clinical; Clinical Research; Clinical Trials; Complication; Cytokine Signaling; Data; Dendritic Cells; Development; Future; Health; Hematopoietic Neoplasms; Hematopoietic Stem Cell Transplantation; Human; IACUC; Immunosuppression; In Vitro; Institutional Review Boards; Interleukin 2 Receptor Gamma; Interleukin-12; Interleukin-2; Interleukin-6; Investigation; JAK2 gene; JAK3 gene; Kinetics; Knowledge; Laboratories; Learning; Leukocytes; Life; Ligation; Liver; MAPK3 gene; Measures; Mediating; Mentorship; Mission; Morbidity - disease rate; Mus; National Heart, Lung, and Blood Institute; Organ; Pathway interactions; Patients; Phenotype; Phosphorylation; Prevention therapy; Prophylactic treatment; Prospective Studies; Protocols documentation; Publishing; Regimen; Regulatory T-Lymphocyte; Research; Risk; STAT3 gene; Sampling Studies; Severities; Signal Transduction; Signaling Protein; Skin; Small Interfering RNA; Stat5 protein; Stem cell transplant; Surrogate Markers; System; T cell response; T-Cell Proliferation; T-Lymphocyte; Testing; Therapeutic Effect; Tissues; Translating; Transplantation; Withdrawal; Work; career; cytokine; design; graft vs host disease; graft vs leukemia effect; in vivo; inhibitor/antagonist; innovation; insight; interleukin-23; lymph nodes; mortality; mouse model; novel; novel strategies; patient population; peripheral blood; prevent; receptor; research study; response; sample collection; small molecule; therapeutic target

DESCRIPTION (provided by applicant): Graft-versus-host disease (GvHD) is a potentially life threatening complication following allogeneic hematopoietic stem cell transplantation (HSCT). JAK2 inhibition induces allotolerance, while preserving Treg development. Preliminary data supports that JAK2 inhibition combined with IL-2 offers a synergistic effect on suppressing alloreactivity. Moreover, blockade of downstream STAT3 abrogates alloresponses as a single agent. These strategies preserve Treg-dependent IL-2/STAT5 signaling, increasing the ratio of STAT5 to STAT3 phosphorylation. The proposed experiments will investigate the influence of JAK2 inhibition paired with IL-2, or STAT3 blockade alone; on the control of allosensitization. This work is highly relevant to the mission of the NHLBI, as it will offer new insights on eliminating the profound morbidity and mortality of GvHD. The proposed experiments will evaluate the following specific aims: Aim 1) Examine JAK2 inhibition paired with IL-2, or STAT3 blockade alone, as a platform to control alloreactivity and optimize Treg expansion in vitro. Aim 2) Investigate the feasibility, biologic influence, and therapeutic effect of skewing the STAT5/STAT3 phosphorylation ratio in a mouse model of GVHD and GVL. Aim 3) Determine if the pSTAT5/pSTAT3 ratio or ERK1/2 phosphorylation in CD4+ or CD8+ T cells on day +21 associate with the development of acute GvHD before day +100. Dendritic cell (DC)-allostimulated T cells will be used to investigate the influence of pSTAT5/pSTAT3 polarization via JAK2 inhibition combined with IL-2, or STAT3 inhibition alone, as a means to suppress alloreactivity and promote Treg expansion. Mechanistic studies will evaluate secondary effects on Treg suppression, STAT signaling, and skewing of T helper subsets and related functions. The resultant data will be confirmed by molecularly targeting STAT3 with siRNA. This concept will be tested in vivo with a GVHD prophylaxis regimen of a JAK2 and IL-2, or STAT3 inhibitor alone, in an MHC- mismatched mouse model (C57BL/6 => Balb/c). Mice will be assessed for GVHD clinical scores, survival, and alterations in Treg/Th1/Th17 subsets. The concept of STAT5/STAT3 and ERK1/2 phosphorylation as biomarkers of impending acute GvHD will be investigated in a 110 patient sample study. JAK2-mediated STAT3 and parallel ERK1/2 phosphorylation will be measured in patients' T cells on day +21 after allogeneic HSCT. The ratio of STAT5/STAT3 phosphorylation will be studied for any protective influence on GvHD. Logical extensions of this work will focus on translating the concept of JAK2 paired with IL-2, or STAT3 blockade alone, in novel clinical trials for GVHD prevention and therapy. The informative data provided by aim 3, if positive, will be validated in a larger patient population to determine f the STAT3, STAT5, and ERK1/2 phosphorylation predict the risk of acute GvHD.

描述（由申请人提供）：在同种异体造血干细胞移植（HSCT）之后，移植物与宿主病（GVHD）是一种潜在的威胁生命并发症的并发症。 JAK2抑制作用诱导同倍耐耐受性，同时保留Treg发育。初步数据支持JAK2抑制与IL-2结合，对抑制同种异体反应性产生了协同作用。此外，下游STAT3的封锁废除了作为单个代理的同种异体。这些策略保留了依赖于Treg的IL-2/STAT5信号传导，从而增加了STAT5与STAT3磷酸化的比率。提出的实验将研究JAK2抑制与IL-2或STAT3封锁的影响；在控制化敏化方面。这项工作与NHLBI的使命高度相关，因为它将为消除GVHD的深刻发病率和死亡率提供新的见解。提出的实验将评估以下特定目的：目标1）检查与IL-2或STAT3封锁配对的JAK2抑制作用，作为控制同种反应性并在体外优化Treg扩展的平台。目标2）研究GVHD和GVL小鼠模型中STAT5/STAT3磷酸化率偏向STAT5/STAT3磷酸化比的可行性，生物学影响和治疗效应。 AIM 3）确定在+21第+100天之前，+21的CD4 +或CD8 + T细胞中的PSTAT5/PSTAT3比或ERK1/2磷酸化是否与急性GVHD的发展相关。树突状细胞（DC）验证的T细胞将用于研究PSTAT5/PSTAT3通过JAK2抑制与IL-2或STAT3抑制相结合的PSTAT5/PSTAT3极化的影响，作为抑制同种反应性和促进TREG膨胀的一种手段。机械研究将评估对T Helper子集和相关功能的Treg抑制，Stat信号传导和偏斜的次要影响。所得数据将通过用siRNA靶向STAT3来确认。该概念将在MHC-不匹配的小鼠模型（C57BL/6 => balb/c）中使用JAK2和IL-2或STAT3抑制剂的GVHD预防方案进行体内测试。将评估小鼠的GVHD临床评分，存活和Treg/Th1/Th17子集的变化。 STAT5/STAT3和ERK1/2磷酸化作为即将发生的急性GVHD的生物标志物的概念将在110项患者样本研究中进行研究。同种异体HSCT后，将在患者的T细胞中测量JAK2介导的STAT3和平行的ERK1/2磷酸化。将研究STAT5/STAT3磷酸化的比率，以确保对GVHD的任何保护作用。这项工作的逻辑扩展将集中于在GVHD预防和治疗的新型临床试验中，将JAK2与IL-2或STAT3封锁配对的概念翻译。 AIM 3提供的信息数据（如果为阳性）将在较大的患者人群中进行验证，以确定STAT3，STAT5和ERK1/2磷酸化的F型磷酸化。