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: 7591371
• 负责人: Margaret S. Bynoe
• 金额: $ 33.69万
• 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/7591371
• 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; Condition; 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; Public Health; 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; 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）是一种中枢神经系统（CNS）炎症性发作，可导致神经元功能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，并且与MS类似，是由促炎性免疫细胞介导的自身免疫性疾病，所述促炎性免疫细胞浸润CNS并产生针对髓鞘组分的免疫应答，导致可引起瘫痪甚至死亡的神经元损伤。CD 73（外-5 '-核苷酸酶）是糖基磷脂酰肌醇（GPI）连接的膜蛋白，其催化AMP的细胞外去磷酸化为腺苷。在免疫反应期间，从受损细胞释放的ATP主要通过CD 73的酶促作用转化为腺苷。这种细胞外腺苷作为一种负反馈信号传递给表达腺苷受体的淋巴细胞，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，CD 73产生的腺苷是淋巴细胞有效迁移到CNS和EAE发展所必需的。本提案的目标是：1）确定腺苷信号传导在调节淋巴细胞迁移到CNS引起自身免疫性疾病中的作用; 2）确定由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。为了实现这些目标，我们将：在野生型和CD 73-/-小鼠中测试EAE诱导之前和期间细胞外腺苷的浓度;研究EAE发展之前和期间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展的后果;确定腺苷受体信号传导缺陷的小鼠是否对EAE诱导敏感;确定腺苷受体信号传导对淋巴细胞进入中枢神经系统的影响，阻断CD 73酶促能力是否可以防止野生型小鼠中EAE的发展或稳定已有EAE的小鼠中的疾病;并研究用PEG-ADA增强腺苷降解是否可以防止野生型小鼠中EAE的发展或稳定疾病。为了确定由腺苷信号调节以影响脉络丛处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丛细胞系来确定脉络丛处的腺苷信号是否可以在体外触发淋巴细胞迁移;我们将在存在或不存在腺苷受体信号的情况下定量体外脉络丛上和EAE期间粘附分子的表达;我们将在存在或不存在腺苷受体信号的情况下，在体内和体外定量脉络丛和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时脉络丛趋化因子分泌的影响。公共卫生相关性：多发性硬化症是一种中枢神经系统慢性炎症性疾病。它是由免疫细胞浸润到中枢神经系统（CNS）介导的，引起炎症，导致神经元损伤，导致瘫痪甚至死亡。EAE是MS的动物模型。我们有证据表明，CD 73产生的腺苷调节淋巴细胞进入EAE小鼠的大脑。这一发现为开发旨在阻断神经炎性疾病（如MS）中CNS的免疫细胞侵袭的疗法提供了潜力。此类疗法可用于阻止MS的进展，甚至可能逆转MS造成的损伤。