CB2 Receptor Regulatin of Inflammatory Response in EAE

描述（由申请人提供）：多发性硬化症（MS）是一种中枢神经系统的慢性脱髓鞘疾病，是西方国家神经功能障碍的主要原因。尽管进行了大量持续的研究工作，现有的治疗方案并不能基本上防止组织损伤和临床残疾。人们越来越认识到多发性硬化症是一种由中枢神经系统炎症攻击引发的自身免疫性神经退行性疾病。我们建议研究一种新合成的分子（O-1966），它可能会显着减少免疫细胞对中枢神经系统损伤的影响。 O-1966 是大麻素类分子的成员。虽然从大麻植物中提取的大麻素已被使用了数千年，但其治疗多发性硬化症等疾病的价值却因其精神活性而受到限制。目前，可以针对主要在免疫细胞上表达的具有免疫调节功能的CB2受体。 O-1966 是一种选择性 CB2 激动剂，可以改变免疫功能，但不会产生精神作用。初步结果表明，刺激 CB2 受体可减少实验性自身免疫性脑脊髓炎 (EAE) 中的中枢神经系统损伤。我们的初步数据表明，O-1966 对 EAE 的保护作用是通过对炎症的影响而发生的。在本提案中，我们将研究 O-1966 特异性影响免疫细胞功能的分子机制。拟议的研究将在分子/细胞和整体动物水平上进行，以便更好地评估 O-1966 的治疗潜力。我们提出选择性 CB2R 激动剂 O-1966 激活 CB2 受体会导致： 1. 通过诱导调节性 T 细胞改变效应 T 细胞的表型； 2.通过减少T细胞滚动和粘附到血管CNS内皮来抑制致脑炎T细胞向CNS的运输。在具体目标 1 中，我们将评估 O-1966 在 EAE 中的作用，并基于 CB2 激活导致 T 细胞分化从致脑炎 Th17/Th1 转变为调节性 T 细胞的假设，表征 CNS 浸润细胞。基于初步研究，我们提出 CB2 选择性激动剂 O-1966 通过诱导耐受性树突状细胞来影响 T 细胞分化。这代表了一个新的研究领域，因为大麻素对 DC 分化的作用尚未得到解决。在具体目标 2 中，我们将通过活体显微镜检查 O-1966 对致脑炎 T 效应器和 Treg 细胞滚动以及 CNS 毛细血管后微静脉粘附的影响，其对 Teff/Treg 细胞和 CNS 内皮细胞中粘附分子表达的影响，以及对参与活化 T 细胞募集到血管周围空间和 CNS 实质的趋化因子表达的影响。 CB2受体配体抗炎作用中分子/细胞因子和机制的鉴定将对中枢神经系统自身免疫/炎症疾病的理解和干预产生重大影响。该提案的最终目标是开发新的有效治疗药物，旨在改变导致多发性硬化症的炎症反应。公共卫生相关性：多发性硬化症是中枢神经系统最常见的慢性脱髓鞘疾病，是西方国家残疾的主要原因。全球大约有 250 万人和 40 万美国人患有多发性硬化症。除了对多发性硬化症患者的生活质量造成经常性的破坏性后果外，其对个人和国家层面的经济影响也是巨大的。据估计，每年治疗多发性硬化症的费用超过25亿美元。尽管进行了大量持续的研究工作，现有的治疗方案并不能基本上防止组织损伤和临床残疾。当前提案的最终目标是开发新的有效治疗药物来治疗多发性硬化症。

DESCRIPTION (provided by applicant): Multiple sclerosis (MS), a chronic demyelinating disease of the central nervous system, is a major cause of neurological disability in Western countries. In spite of intensive and sustained research efforts, existing treatment options do not substantially prevent tissue damage and clinical disability. MS is increasingly recognized as an autoimmune neurodegenerative disease triggered by inflammatory attacks of the CNS. We propose to study a newly synthesized molecule (O-1966) that may significantly reduce the contribution of immune cells to CNS damage. O-1966 is a member of the cannabinoid class of molecules. While cannabinoids derived from the plant Cannabis Sativa have been used for thousands of years, their value in the treatment of diseases such as MS has been limited by their psychoactive properties. Presently, it is possible to target the CB2 receptors, primarily expressed on immune cells, which exhibit immunomodulatory functions. O-1966, a selective CB2 agonist, alters immune functions without psychoactive effects. Preliminary results showed that stimulation of CB2 receptors decreases CNS damage in experimental autoimmune encephalomyelitis (EAE). Our preliminary data suggest that the protective action of O-1966 in EAE occurs through effects on inflammation. In this proposal we will investigate the molecular mechanisms by which O-1966 specifically influences the function of immune cells. The proposed studies will be conducted at both molecular/cellular and whole animal level, to allow a better evaluation of the therapeutic potential of O- 1966. We propose that CB2 receptor activation by the selective CB2R agonist O-1966 results in: 1. changes in the phenotype of effector T cells through the induction of regulatory T cells; and 2. inhibition of encephalitogenic T cell traffic to the CNS through a reduction in T cell rolling and adhesion to the vascular CNS endothelium. In Specific Aim 1 we will evaluate the effect of O-1966 in EAE and characterize the CNS infiltrating cells, based on the hypothesis that CB2 activation results in a shift in T cell differentiation from encephalitogenic Th17/Th1 to regulatory T cells. Based on preliminary studies, we propose that the CB2 selective agonist O-1966 affects T cell differentiation through the induction of tolerogenic dendritic cells. This represents a new research area, since the role of cannabinoids on DC differentiation has not been addressed. In Specific Aim 2 we will examine the effects of O-1966 on encephalitogenic Teffector and Treg cell rolling and adhesion to CNS postcapillary venules by intravital microscopy, its effect on the expression of adhesion molecules in Teff/Treg cells and in CNS endothelial cells, and on the expression of chemokines involved in the recruitment of activated T cells to the perivascular space and CNS parenchyma. The identification of molecular/cellular factors and of the mechanisms involved in the anti-inflammatory effect of CB2 receptor ligands will have a significant impact on the understanding and intervention in CNS autoimmune/inflammatory diseases. The ultimate goal of this proposal is the development of new effective therapeutic agents targeted at modification of the inflammatory responses that contribute to MS. PUBLIC HEALTH RELEVANCE: Multiple sclerosis, the most common chronic demyelinating disease of the central nervous system, is a major cause of disability in Western countries. Approximately 2.5 million people worldwide and 400,000 Americans suffer from MS. In addition to the often devastating consequences for quality of life for individuals suffering from MS, the economic impact at both the personal and national level is substantial. It is estimated that the annual costs for the treatment of MS is in excess of $2.5 billion. In spite of intensive and sustained research efforts, existing treatment options do not substantially prevent tissue damage and clinical disability. The ultimate goal of the current proposal is the development of new effective therapeutic agents for the treatment of MS.