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慢性脱髓鞘疾病，是西方国家神经系统残疾的主要原因。尽管进行了深入和持续的研究，但现有的治疗方案并不能实质上防止组织损伤和临床残疾。MS越来越多地被认为是由CNS的炎性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CNS损伤的贡献。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经使用了数千年，但它们在治疗MS等疾病方面的价值受到其精神活性的限制。目前，有可能靶向主要在免疫细胞上表达的CB 2受体，其表现出免疫调节功能。O-1966是一种选择性CB 2激动剂，可改变免疫功能，但无精神活性作用。初步结果表明，CB 2受体的刺激减少了实验性自身免疫性脑脊髓炎（EAE）的CNS损伤。我们的初步数据表明，O-1966在EAE中的保护作用是通过对炎症的影响而发生的。在这项提议中，我们将研究O-1966特异性影响免疫细胞功能的分子机制。拟定的研究将在分子/细胞和整个动物水平进行，以便更好地评价O- 1966的治疗潜力。我们提出，CB 2受体激活的选择性CB 2 R激动剂O-1966的结果：1。通过诱导调节性T细胞改变效应T细胞的表型;和2.通过减少T细胞滚动和粘附到血管CNS内皮来抑制致脑炎性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CB 2活化导致T细胞分化从致脑炎性Th 17/Th 1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CB 2选择性激动剂O-1966通过诱导致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。在特定目标2中，我们将通过活体显微镜检查O-1966对致脑炎性T效应细胞和Treg细胞滚动和粘附至CNS毛细血管后小静脉的影响，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细胞向血管周围空间和CNS实质募集的趋化因子表达的影响。识别分子/细胞因子和参与CB 2受体配体抗炎作用的机制将对CNS自身免疫性/炎症性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剂，其目标是改变导致MS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。全世界约有250万人和40万美国人患有MS。除了对患有MS的个人的生活质量通常造成破坏性后果外，对个人和国家层面的经济影响也是巨大的。据估计，每年用于治疗MS的费用超过25亿美元。尽管进行了深入和持续的研究，但现有的治疗方案并不能实质上防止组织损伤和临床残疾。本提案的最终目标是开发用于治疗MS的新的有效治疗剂。

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.