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

帕金森病(PD)是一种进行性退行性疾病，影响着近8万退伍军人。既然有 没有治愈方法，必须开发新的疗法来阻止疾病的发展。而这种脱根的机制- 发病过程仍然难以捉摸，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，在脊髓(SC)退变中起关键作用，并可能是驱动因素 炎症和疾病进展。来自MPTP小鼠和人类PD样本的初步数据显示 CD4+和CD8+T细胞向SC和SN的渗透，增加小鼠脾细胞中的CD4+T细胞，以及 血清细胞因子/趋化因子升高。检测到扩增的细胞毒性CD4+T细胞亚群 MPTP小鼠和DSP4/6OHDA诱导的大鼠脾细胞。钙肽蛋白(钙蛋白酶抑制剂)治疗 在MPTP小鼠中取消了这种CD4+亚型，表明Calain在T细胞激活和CD4+亚型中的作用 可能在炎症过程中起关键作用。趋化因子受体CCR-1是一种钙离子激动剂，重要的是， 可以激活钙蛋白酶。由于Ccr-1配体(MIP-1α，RANTES)促进T细胞在SC中的运输，它们在 对炎症反应进行评估。在用钙肽治疗MPTP小鼠后，它们的数量显著减少， 行为功能明显改善。帕金森病患者的SC显示小胶质细胞活化和 星形胶质细胞、CD_4~+/CD_8~+T细胞的浸润和钙蛋白的增加。对原代小胶质细胞激活的抑制作用 钙肽蛋白和CCR-1拮抗剂(BX471)诱导的钙离子载体显著降低细胞因子/ 趋化因子，提示它们作为治疗MPTP诱导的神经毒性的药物的潜力。因此，我们 假设Calain激活，炎性T细胞(Th1/Th17，CD8+)的浸润和释放 细胞因子/趋化因子参与帕金森病的进行性退变，以及钙蛋白酶抑制剂和CCR-1 拮抗剂治疗可以减少退行性变，减缓疾病进展，改善功能。三 具体目标是：(1)研究钙蛋白酶调节和T细胞浸润在SC退变中的作用。 MPTP小鼠的发病和疾病进展，表征浸润性T细胞，评估细胞因子/趋化因子水平 测定血清中细胞死亡参数和钙蛋白酶活性；(2)检测T细胞和 帕金森病患者SC中小胶质细胞的激活，T细胞亚群的特征，并与特异性 目的1、测定PD患者血清中的趋化因子/细胞因子；(3)检测治疗前后PD患者血清中趋化因子/细胞因子的变化。 应用钙蛋白酶抑制剂(CalPeptin，SNJ1945)和BX471的MPTP小鼠单独或联合应用可减少炎症反应。 运动事件与功能退化和改善，与DigiGait分析相关，并检查是否 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，并可通过 钙肽蛋白和BX471。在MPTP小鼠中发现的一种CD4亚型的关键发现也将在鱼藤酮和 DSP4/60OHDA大鼠模型。我们的目标是开发PD治疗的策略，使用阻断炎症的药物。 运动过程，保护神经元，控制疾病进展，改善功能。

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.