Attenuation of Inflammatory Response in Progressive Neurodegeneration in Parkinson's Disease

帕金森病进行性神经变性中炎症反应的减弱

• 批准号: 10158428
• 负责人: NAREN L BANIK
• 金额: --
• 依托单位: RALPH H JOHNSON VA MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-04-01 至 2023-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10158428
• 关键词: 1-Methyl-4-phenylpyridinium; Affect; Astrocytes; Attenuated; Autopsy; Behavioral; Biological Assay; Bradykinesia; Brain; CASP1 gene; CCL2 gene; CCR1 gene; CD4 Positive T Lymphocytes; CD8-Positive T-Lymphocytes; CD8B1 gene; Calcium; Calpain; Caspase; Cell Death; Cells; Characteristics; Chemotaxis; Chronic; DSP 4; Data; Degenerative Disorder; Disease; Disease Progression; Elderly; Enzyme-Linked Immunosorbent Assay; Event; Flow Cytometry; Gliosis; Goals; Human; Immunofluorescence Immunologic; In Situ Nick-End Labeling; In Vitro; Infiltration; Inflammation; Inflammatory; Inflammatory Response; Interleukin-1 beta; Interleukin-12; Invaded; Ionophores; Laboratories; Levodopa; Lewy Bodies; Ligands; MPTP model; MPTP mouse; MPTP treatment; Methods; Microglia; Modeling; Motor Activity; Motor Neurons; Movement Disorders; Mus; Nerve Degeneration; Neurodegenerative Disorders; Neurons; Oxidopamine; Parkinson Disease; Pathogenesis; Patients; Play; Process; Production; Proteins; Publishing; RANTES; Rattus; Regulation; Role; Rotenone; Sampling; Serum; Small Interfering RNA; Spinal Cord; Spinal cord damage; Spleen; Splenocyte; Substantia nigra structure; T cell regulation; T-Cell Activation; T-Lymphocyte; TNF gene; Testing; Therapeutic; Tin; Tissues; Tremor; Tumor-infiltrating immune cells; Veterans; Western Blotting; alpha synuclein; attenuation; calpain inhibitor; cell motility; chemokine; chemokine receptor; cytokine; cytotoxic; disorder control; dopaminergic neuron; glial activation; improved; improved functioning; knock-down; migration; neuron loss; neurotoxicity; novel therapeutics; prevent; progressive neurodegeneration; response; synuclein; tissue degeneration; trafficking

Parkinson's disease (PD), a progressive degenerative disorder, affects almost 80,000 Veterans. Since there is no cure, new therapies must be developed to halt disease progression. While the mechanisms of this degen- erative process remain elusive, chronic inflammation may be involved. Calpain activates microglia and T cells, induces T cell migration/chemotaxis, plays a pivotal role in spinal cord (SC) degeneration, and may be a driver of inflammation and disease progression. Preliminary data from MPTP mice and human PD samples showed infiltration of CD4+ and CD8+ T cells into SC and SN, increased CD4+ T cells in mouse splenocytes, and elevated serum cytokines/chemokines. An expanded subpopulation of cytotoxic CD4+ T cells was detected in splenocytes from MPTP mice and DSP-4/6OHDA-induced rats. Calpeptin (calpain inhibitor) treatment abolished this CD4+ subtype in MPTP mice, suggesting calpain's role in T cell activation and the CD4+ subtype may be critical in the inflammatory process. Chemokine receptor CCR-1 is a Ca2+ mobilizer and, importantly, can activate calpain. Since CCR-1 ligands (MIP-1α, RANTES) promote trafficking of T cells in SC, their role in inflammation was assessed. They were significantly reduced following treatment of MPTP mice with calpeptin, and behavioral function was remarkably improved. SC from PD patients revealed activation of microglia and astrocytes, infiltration of CD4+/CD8+ T cells, and increased calpain. Inhibition of primary microglia activation by Ca2+ ionophore by calpeptin and CCR-1 antagonist (BX471) produced marked reduction in cytokines/ chemokines, suggesting their potential as agents for treatment of MPTP-induced neurotoxicity. Thus, we hypothesize that calpain activation, infiltration of inflammatory T cells (Th1/Th17, CD8+), and released cytokines/chemokines are involved in progressive degeneration in PD, and calpain inhibitor and CCR-1 antagonist treatment may reduce degeneration, slow disease progression, and improve function. Three specific aims are proposed: (1) investigate the role of calpain regulation and T cell infiltration in SC degenera- tion and disease progression in MPTP mice, characterize infiltrating T cells, assess cytokine/chemokine levels in sera, and determine cell death parameters and calpain activation in SC; (2) examine infiltration of T cells and activation of microglia in SC from PD patients, characterize T cell subpopulations, and correlate with Specific Aim 1, determine chemokines/cytokines in serum from PD patients; and (3) examine whether treatment of MPTP mice with calpain inhibitors (calpeptin, SNJ1945) and BX471 alone or in combination will reduce inflam- matory events and degeneration and improve function, correlate with DigiGait analysis, and examine whether primary microglia activated by calcium ionophore upregulate chemokines/cytokines and can be controlled by calpeptin and BX471. The key finding of a CD4-subtype in MPTP mice will also be verified in rotenone and DSP-4/6OHDA rat models. The goal is to develop strategies for PD therapy with agents that block the inflam- matory process, protect neurons, control disease progression, and improve function.

帕金森病 (PD) 是一种进行性退行性疾病，影响着近 80,000 名退伍军人。既然有 无法治愈，必须开发新疗法来阻止疾病进展。虽然这种退化的机制 生成过程仍然难以捉摸，可能涉及慢性炎症。钙蛋白酶激活小胶质细胞和 T 细胞， 诱导 T 细胞迁移/趋化性，在脊髓 (SC) 变性中发挥关键作用，并且可能是驱动因素 炎症和疾病进展。 MPTP 小鼠和人类 PD 样本的初步数据显示 CD4+ 和 CD8+ T 细胞浸润 SC 和 SN，小鼠脾细胞中 CD4+ T 细胞增加， 血清细胞因子/趋化因子升高。检测到细胞毒性 CD4+ T 细胞的扩大亚群 来自 MPTP 小鼠和 DSP-4/6OHDA 诱导的大鼠的脾细胞。 Calpeptin（钙蛋白酶抑制剂）治疗 在 MPTP 小鼠中消除了这种 CD4+ 亚型，表明钙蛋白酶在 T 细胞激活和 CD4+ 亚型中的作用 可能在炎症过程中至关重要。趋化因子受体 CCR-1 是一种 Ca2+ 动员剂，重要的是， 可以激活钙蛋白酶。由于 CCR-1 配体（MIP-1α、RANTES）促进 SC 中 T 细胞的运输，因此它们在 评估炎症。用 calpeptin 处理 MPTP 小鼠后，它们显着减少， 行为功能显着改善。 PD 患者的 SC 揭示了小胶质细胞的激活 星形胶质细胞、CD4+/CD8+ T 细胞浸润以及钙蛋白酶增加。抑制原代小胶质细胞活化 Calpeptin 和 CCR-1 拮抗剂 (BX471) 产生的 Ca2+ 离子载体显着减少细胞因子/ 趋化因子，表明它们作为治疗 MPTP 诱导的神经毒性的药物的潜力。因此，我们 假设钙蛋白酶激活、炎症 T 细胞（Th1/Th17、CD8+）浸润并释放 细胞因子/趋化因子参与 PD 进行性退化，钙蛋白酶抑制剂和 CCR-1 拮抗剂治疗可以减少变性、减缓疾病进展并改善功能。三 提出了具体目标：（1）研究钙蛋白酶调节和 T 细胞浸润在 SC 退化中的作用 MPTP 小鼠的免疫和疾病进展，表征浸润 T 细胞，评估细胞因子/趋化因子水平 在血清中，并测定 SC 中的细胞死亡参数和钙蛋白酶激活； (2)检查T细胞的浸润情况 PD 患者 SC 中小胶质细胞的激活，表征 T 细胞亚群，并与特异性相关 目标1，测定PD患者血清中的趋化因子/细胞因子； (3) 检查是否治疗 MPTP 小鼠单独使用钙蛋白酶抑制剂（calpeptin，SNJ1945）和 BX471 或联合使用可减少炎症 发生事件和退化并改善功能，与 DigiGait 分析相关联，并检查是否 由钙离子载体激活的原代小胶质细胞上调趋化因子/细胞因子，并且可以通过以下方式控制 钙肽和BX471。 MPTP 小鼠中 CD4 亚型的关键发现也将在鱼藤酮和 DSP-4/6OHDA 大鼠模型。我们的目标是制定 PD 治疗策略，使用阻断炎症的药物。 生过程，保护神经元，控制疾病进展，并改善功能。