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.

帕金森氏病（PD）是一种进行性退行性疾病，影响了近80,000名退伍军人。因为有 不能治愈，必须开发新的疗法来阻止疾病进展。而这种degen的机制 erication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）变性中起关键作用，可能是驱动器 炎症和疾病进展。来自MPTP小鼠和人类PD样品的初步数据 CD4+和CD8+ T细胞浸润到SC和SN中，小鼠脾细胞中的CD4+ T细胞增加，并且 血清细胞因子/趋化因子升高。在 来自MPTP小鼠和DSP-4/6OHDA诱导的大鼠的脾细胞。钙肽蛋白（钙蛋白酶抑制剂）治疗 在MPTP小鼠中废除了这种CD4+亚型，这表明Calpain在T细胞激活和CD4+亚型中的作用 在炎症过程中可能至关重要。趋化因子受体CCR-1是CA2+动员，重要的是 可以激活钙蛋白酶。由于CCR-1配体（MIP-1α，Rantes）促进了T细胞在SC中的运输，因此它们在 评估炎症。用Calpeptin治疗MPTP小鼠后，它们大大降低了 行为功能得到了极大的改善。 PD患者的SC显示小胶质细胞激活和 星形胶质细胞，CD4+/CD8+ T细胞的浸润，并增加钙蛋白酶。通过抑制原发性小胶质细胞激活 CA2+离子载体由Calpeptin和CCR-1拮抗剂（BX471）产生的细胞因子/ 趋化因子，表明它们作为治疗MPTP诱导神经毒性的药物的潜力。那，我们 假设钙蛋白酶激活，炎性T细胞的浸润（TH1/TH17，CD8+），并释放 细胞因子/趋化因子参与PD进行性变性，CALPAIN抑制剂和CCR-1 拮抗剂治疗可以减少变性，疾病进展缓慢并改善功能。三 提出了具体目的：（1）研究钙蛋白酶调节和T细胞浸润在SC degenera-的作用 MPTP小鼠的疾病和疾病进展，表征浸润T细胞，评估细胞因子/趋化因子水平 在血清中，并确定SC中的细胞死亡参数和钙蛋白酶激活； （2）检查T细胞和 PD患者SC中的小胶质细胞激活，表征T细胞亚群，并与特定相关 AIM 1，确定PD患者血清中的趋化因子/细胞因子； （3）检查是否治疗 使用钙蛋白酶抑制剂（Calpeptin，SNJ1945）和BX471或组合使用的MPTP小鼠将减少毒气 机器事件以及变性和改进功能，与数字观念分析相关，并检查是否是否 由钙离子载体激活的一级小胶质细胞上调趋化因子/细胞因子，可以由 Calpeptin和BX471。 MPTP小鼠中CD4-Subtype的关键发现也将在鱼藤酮和 DSP-4/6OHDA大鼠模型。目的是通过使用阻止Inferm-的药物制定PD治疗策略 机械过程，保护神经元，控制疾病进展并改善功能。