CD-73 adenosine signaling in the central nervous system in disease and health

疾病和健康中枢神经系统中的 CD-73 腺苷信号转导

• 批准号: 8019302
• 负责人: Margaret S. Bynoe
• 金额: $ 3.17万
• 依托单位: CORNELL UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-09-01 至 2013-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8019302
• 关键词: 5' -Nucleotidase; Adenosine; Adenosine A2A Receptor; Adoptive Transfer; Animal Model; Attention; Autoimmune Diseases; Autoimmune Responses; Blood; Blood - brain barrier anatomy; Brain; CD4 Positive T Lymphocytes; Cell Adhesion Molecules; Cell Line; Cells; Central Nervous System Diseases; Cerebrospinal Fluid; Cessation of life; Chronic; Development; Disease; Encephalitis; Encephalomyelitis; Experimental Autoimmune Encephalomyelitis; Feedback; Glycosylphosphatidylinositols; Goals; Health; Immune; Immune response; Immunity; Immunologic Monitoring; In Vitro; Infection; Infiltration; Inflammation; Inflammatory; Lead; Link; Lymphocyte; Mediating; Mediator of activation protein; Membrane Proteins; Multiple Sclerosis; Mus; Myelin; Neuraxis; Neurons; Paralysed; Pattern; Process; Protein Dephosphorylation; Purinergic P1 Receptors; Receptor Signaling; Regulation; Resistance; Role; Signal Transduction; Staining method; Stains; Structure of choroid plexus; T-Lymphocyte; Testing; Time; Vascular blood supply; Wild Type Mouse; base; cell injury; chemokine; extracellular; in vivo; migration; mouse model; neuron loss; pegademase bovine; prevent; public health relevance; therapy development

DESCRIPTION (provided by applicant): Multiple Sclerosis (MS) is an inflammatory attack on the central nervous system (CNS) that results in loss of neuronal function and can lead to death. Experimental Autoimmune Encephalomyelitis (EAE) is a mouse model for MS, and like MS, is an autoimmune disease mediated by proinflammatory immune cells that infiltrate the CNS and mount immune responses against myelin components, resulting in neuronal damage that can cause paralysis and even death. CD73 (Ecto-5'-nucleotidase) is a glycosyl phosphatidylinositol (GPI)-linked membrane protein that catalyzes the extracellular dephosphorylation of AMP to adenosine. During an immune response, ATP released from damaged cells is converted to adenosine, mainly through the enzymatic action of CD73. This extracellular adenosine acts as a negative feedback signal to lymphocytes that express adenosine receptors and essentially turns off the immune response to prevent excessive cellular damage. Preliminary evidence shows CD73 generated adenosine is required for efficient lymphocyte migration into the CNS and for EAE development. The goals of this proposal are to 1) determine the role of adenosine signaling in regulating lymphocyte migration into the CNS to cause autoimmune disease; and 2) define the adhesion molecules and chemokines that are regulated by adenosine signaling to influence lymphocyte migration at the choroid plexus. To achieve these goals we will: test the concentration of extracellular adenosine before and during EAE induction in wild type and CD73-/- mice; investigate the expression pattern and level of adenosine receptors in the brain before and during EAE development; determine the consequences of adenosine receptor blockade during EAE on lymphocyte infiltration and disease development; determine whether mice deficient in adenosine receptor signaling are susceptible to EAE induction; determine the effect of adenosine receptor signaling on lymphocyte passage into the CNS, whether blockade of CD73 enzymatic ability can prevent development of EAE in wild type mice or stabilize disease in mice with preexisting EAE; and investigate whether enhancing adenosine degradation with PEG-ADA can prevent development of EAE in wild type mice or stabilize the disease. To define the adhesion molecules and chemokines that are regulated by adenosine signaling to influence lymphocyte migration at the choroid plexus, we will use a mouse choroid plexus cell line to determine whether adenosine signaling at the choroid plexus can trigger lymphocyte migration in vitro; we will quantify the expression of adhesion molecules on the choroid plexus in vitro and during EAE in the presence or absence of adenosine receptor signaling; we will quantify the expression of adhesion molecules on the choroid plexus and lymphocytes in vivo and in vitro in the presence or absence of adenosine receptor signaling. Finally, we will investigate the impact of adenosine receptor signaling on chemokine secretion at the choroid plexus during EAE. PUBLIC HEALTH RELEVANCE: Multiple sclerosis is a chronic inflammatory disease of the central nervous system. It is mediated by infiltration of immune cells into the central nervous system (CNS), causing inflammation which results in damage to neurons, leading to paralysis and even death. EAE is the animal model for MS. We have evidence suggesting that CD73-generated adenosine regulates the entry of lymphocytes into the brains of mice with EAE. This finding provides potential for development of therapies aimed at blocking immune cell invasion of the CNS in neuroinflammatory diseases such as MS. Such therapies could be used to stop the progress of, or even possibly reverse the damage caused by MS.

描述（由申请人提供）：多发性硬化症（MS）是对中枢神经系统（CNS）的炎症攻击，导致神经元功能的丧失并可能导致死亡。实验性自身免疫性脑脊髓炎（EAE）是MS的小鼠模型，与MS一样，是一种由促炎性免疫细胞介导的自身免疫性疾病，可浸润CNS并针对髓磷脂成分进行免疫反应，导致神经元损伤，导致可能导致麻痹和甚至死亡。 CD73（ECTO-5'-核苷酸酶）是一种糖基磷脂酰肌醇（GPI）连接的膜蛋白，可催化AMP的细胞外去磷酸化为腺苷。在免疫反应过程中，主要通过CD73的酶促作用转化为从受损细胞释放的ATP转化为腺苷。这种细胞外腺苷充当表达腺苷受体的淋巴细胞的负反馈信号，并基本上关闭免疫反应以防止过度的细胞损伤。初步证据表明，有效淋巴细胞迁移到中枢神经系统和EAE发育中需要CD73产生的CD73。该提案的目标是1）确定腺苷信号传导在调节淋巴细胞迁移到中枢神经系统中的作用，以引起自身免疫性疾病； 2）定义粘附分子和趋化因子受腺苷信号传导调节以影响脉络丛处淋巴细胞迁移。为了实现这些目标，我们将：测试野生型和CD73 - / - 小鼠EAE诱导之前和期间细胞外腺苷的浓度；在EAE发育前后，研究大脑中腺苷受体的表达模式和水平；确定EAE期间腺苷受体阻断对淋巴细胞浸润和疾病发育的后果；确定缺乏腺苷受体信号传导的小鼠是否易于EAE诱导；确定腺苷受体信号传导对CNS淋巴细胞传递的影响，CD73酶促能力是否可以防止野生型小鼠中EAE的发育或以前存在EAE的小鼠稳定疾病；并研究用钉蛋白酶增强腺苷降解是否可以防止野生型小鼠的EAE发展或稳定疾病。为了定义腺苷信号传导调节以影响脉络丛淋巴细胞迁移的粘附分子和趋化因子，我们将使用小鼠脉络膜丛细胞系来确定脉络膜上的腺苷信号传导是否可以触发淋巴细胞的迁移。在存在或不存在腺苷受体信号传导的情况下，我们将在体外和EAE期间量化粘附丛上粘附分子的表达；在存在或不存在腺苷受体信号传导的情况下，我们将量化体内和体外淋巴细胞上粘附分子的表达。最后，我们将研究腺苷受体信号传导对EAE期间脉络丛趋化因子分泌的影响。公共卫生相关性：多发性硬化症是中枢神经系统的慢性炎症性疾病。它是通过免疫细胞浸润到中枢神经系统（CNS）的介导的，导致炎症导致神经元损害，导致瘫痪甚至死亡。 EAE是MS的动物模型。我们有证据表明，CD73生成的腺苷调节淋巴细胞进入小鼠的大脑中，用EAE进入小鼠的大脑。这一发现为开发旨在阻止神经炎症性疾病（如MS）中CNS的免疫细胞侵袭的疗法的发展提供了潜力。这种疗法可用于阻止MS造成的损害的进度，甚至可能逆转。