Cannabinoid-mediated mitigation of graft versus host disease: Roles of CB2 receptors and adenosine signaling

大麻素介导的移植物抗宿主病缓解：CB2 受体和腺苷信号传导的作用

基本信息

批准号：
9402352
负责人：
William R. Drobyski
金额：
$ 53.87万
依托单位：
MEDICAL COLLEGE OF WISCONSIN
依托单位国家：
美国
项目类别：
财政年份：
2017
资助国家：
美国
起止时间：
2017-08-01 至 2021-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9402352
关键词：
2-arachidonylglycerol ADORA2A gene Acute Acute Graft Versus Host Disease Address Adenosine Agonist Allogenic Anti-Inflammatory Agents Anti-inflammatory Biology Bone Marrow Transplantation CNR2 gene Calcineurin Cannabidiol Cannabinoids Cannabis Cells Chronic Phase Chronic Phase of Disease Clinical Complication Development Disease FOXP3 gene Fibrosis Fostering Functional disorder Gastrointestinal tract structure Genetic Goals Hematopoietic Hematopoietic Stem Cell Transplantation Immune Inflammation Inflammatory Lead Ligands Liver Mediating Morbidity - disease rate Organ Pathogenesis Pathway interactions Patients Pharmacology Phase Play Population Production Purinergic P1 Receptors Receptor Signaling Regulatory T-Lymphocyte Reporter Role Severities Severity of illness Signal Pathway Signal Transduction Site Skin Stem cell transplant Syndrome Testing Time Transplant Recipients base chronic graft versus host disease clinically relevant cytokine design experimental study graft vs host disease insight mortality mouse model novel novel strategies phytocannabinoid preclinical study prevent receptor receptor expression reconstitution stem treatment strategy

项目摘要

Graft versus host disease (GVHD) is the major complication associated with allogeneic hematopoietic stem cell transplantation, and is characterized by both acute and chronic phases which have different pathophysiological mechanisms. Despite the use of pharmacological strategies which are primarily based on the administration of calcineurin-based agents, both acute and chronic GVHD remain major clinical problems. In preliminary studies, we have identified that signaling through the cannabinoid type 2 receptor (CB2R) mitigates the severity of both acute GVHD and also plays a pivotal role in reducing fibrosis in chronic GVHD. Moreover, we have discovered that that the nonpsychoactive, phytocannabinoid, cannabidiol (CBD) also inhibits GVH reactivity and potentiates adenosine signaling. Based on these studies, our overall hypothesis is that cannabinoids mitigate the severity of both acute and chronic GVHD, and that these anti-inflammatory effects are mechanistically mediated through the CB2R and the adenosine receptor signaling pathways. Studies in Specific Aim 1 will utilize a novel CB2R reporter mouse model that has flanking lox P sites that will allow us to both identify the critical immune cell populations that express the CB2R, and then eliminate these identified cells to formally test a functional role for these cells in acute GVHD biology. We will also employ several clinically relevant pharmacological approaches to determine whether administration of direct and selective CB2R agonists or, alternatively agents that inhibit the degradation of 2-arachidonyl glycerol (2-AG) which is the major endogenous ligand for the CB2R, can reduce the severity of this disease. Studies in Specific Aim 2 will define the role of CBD in mitigating the severity of acute GVHD by using selective pharmacological antagonists and genetic strategies to test the hypothesis that CBD regulates GVHD through the adenosine 2A receptor (A2AR) signaling pathway. We will also examine whether CBD augments the reconstitution of regulatory T cells and stabilizes Foxp3 expression in these cells through adenosine signaling. Experiments in Specific Aim 3 will determine whether cannabinoid signaling prevents the development of chronic GVHD-associated fibrosis which is a hallmark of this disease. Specifically, we will define the critical CB2R expressing immune cell populations which are present during the fibrotic phase of chronic GVHD and use cell-specific Cre-lox deletion approaches to define the functional significance of CB2R expression in identified cells. We will also assess whether either pharmacological administration of direct CB2R agonists, agents that inhibit the degradation of 2-AG, and CBD can prevent chronic GVHD-associated fibrosis. The overall goal of these studies is to define the mechanisms by which cannabinoids modulate both acute and chronic GVHD, and to test clinically relevant strategies that are designed to mitigate these complications in allogeneic hematopoietic stem cell transplant recipients.

移植物抗宿主病（GVHD）是同种异体造血干相关的主要并发症细胞移植，具有急性期和慢性期的特点，其具有不同的病理生理机制。尽管使用主要基于在基于钙调神经磷酸酶的药物的施用方面，急性和慢性 GVHD 仍然是临床上的主要问题问题。在初步研究中，我们已经确定通过 2 型大麻素发出信号受体 (CB2R) 可减轻急性 GVHD 的严重程度，并在减少纤维化方面发挥关键作用在慢性 GVHD 中。此外，我们发现非精神活性植物大麻素大麻二酚 (CBD) 还可抑制 GVH 反应性并增强腺苷信号传导。基于这些研究中，我们的总体假设是大麻素可以减轻急性和慢性疾病的严重程度慢性 GVHD，并且这些抗炎作用是机械介导的通过CB2R和腺苷受体信号通路。具体目标研究 1 将利用一种新型 CB2R 报告小鼠模型，该模型具有侧翼 lox P 位点，使我们能够识别表达 CB2R 的关键免疫细胞群，然后消除这些识别的细胞细胞正式测试这些细胞在急性 GVHD 生物学中的功能作用。我们还将聘请数名临床相关的药理学方法来确定是否直接或选择性给药 CB2R 激动剂或抑制 2-花生四烯酸甘油 (2-AG) 降解的药物，是CB2R的主要内源性配体，可以减轻这种疾病的严重程度。具体研究目标 2 将定义 CBD 通过选择性使用来减轻急性 GVHD 严重程度的作用药理学拮抗剂和遗传策略来检验 CBD 调节 GVHD 的假设通过腺苷2A受体（A2AR）信号通路。我们还将检查 CBD 是否通过增强调节性 T 细胞的重建并稳定这些细胞中的 Foxp3 表达腺苷信号传导。具体目标 3 中的实验将确定大麻素信号传导是否有效防止慢性 GVHD 相关纤维化的发展，这是该疾病的标志。具体来说，我们将定义关键的 CB2R 表达免疫细胞群，这些细胞群存在于慢性 GVHD 的纤维化阶段，并使用细胞特异性 Cre-lox 缺失方法来定义 CB2R 表达在已识别细胞中的功能意义。我们还将评估是否直接CB2R激动剂、抑制2-AG降解的药物的药理给药，以及 CBD 可以预防慢性 GVHD 相关纤维化。这些研究的总体目标是定义大麻素调节急性和慢性 GVHD 的机制，并测试临床相关性旨在减轻同种异体造血干细胞这些并发症的策略移植受者。