Ly Diane, Dongol Anjila, Cuthbertson Peter, Guy Thomas V, Geraghty Nicholas J, Sophocleous Reece A, Sin Lucia, Turner Bradley J, Watson Debbie, Yerbury Justin J, Sluyter Ronald
Illawarra Health and Medical Research Institute, Wollongong, NSW, 2522, Australia.
Molecular Horizons and School of Chemistry and Molecular Bioscience, University of Wollongong, Wollongong, NSW, 2522, Australia.
Purinergic Signal. 2020 Mar;16(1):109-122. doi: 10.1007/s11302-020-09692-4. Epub 2020 Mar 13.
The ATP-gated P2X7 ion channel has emerging roles in amyotrophic lateral sclerosis (ALS) progression. Pharmacological blockade of P2X7 with Brilliant Blue G can ameliorate disease in SOD1 mice, but recent data suggests that this antagonist displays poor penetration of the central nervous system (CNS). Therefore, the current study aimed to determine whether the CNS-penetrant P2X7 antagonist, JNJ-47965567, could ameliorate ALS progression in SOD1 mice. A flow cytometric assay revealed that JNJ-47965567 impaired ATP-induced cation dye uptake in a concentration-dependent manner in murine J774 macrophages. Female and male SOD1 mice were injected intraperitoneally with JNJ-47965567 (30 mg/kg) or 2-(hydroxypropyl)-beta-cyclodextrin (vehicle control) three times a week from disease onset until end stage, when tissues were collected and studied. JNJ-47965567 did not impact weight loss, clinical score, motor (rotarod) coordination or survival compared to control mice. NanoString analysis revealed altered spinal cord gene expression in JNJ-47965567 mice compared to control mice, but such differences were not confirmed by quantitative PCR. Flow cytometric analyses revealed no differences between treatments in the frequencies or activation status of T cell or dendritic cell subsets in lymphoid tissues or in the concentrations of serum cytokines. Notably, serum IL-27, IFNβ and IL-10 were present in relatively high concentrations compared to other cytokines in both groups. In conclusion, JNJ-47965567 administered thrice weekly from disease onset did not alter disease progression or molecular and cellular parameters in SOD1 mice.
ATP门控的P2X7离子通道在肌萎缩侧索硬化症（ALS）进展中发挥着新出现的作用。用亮蓝G对P2X7进行药理阻断可改善SOD1小鼠的病情，但最近的数据表明，这种拮抗剂对中枢神经系统（CNS）的穿透力较差。因此，本研究旨在确定具有CNS穿透性的P2X7拮抗剂JNJ-47965567是否能改善SOD1小鼠的ALS进展。流式细胞术分析显示，JNJ-47965567在鼠J774巨噬细胞中以浓度依赖性方式损害ATP诱导的阳离子染料摄取。从疾病发作到终末期，每周三次给雌性和雄性SOD1小鼠腹腔注射JNJ-47965567（30mg/kg）或2-（羟丙基）-β-环糊精（溶剂对照），之后收集组织并进行研究。与对照小鼠相比，JNJ-47965567对体重减轻、临床评分、运动（转棒）协调性或存活率没有影响。NanoString分析显示，与对照小鼠相比，JNJ-47965567处理的小鼠脊髓基因表达发生了改变，但定量PCR未证实这些差异。流式细胞术分析显示，各处理组在淋巴组织中T细胞或树突状细胞亚群的频率或激活状态或血清细胞因子浓度方面没有差异。值得注意的是，与两组中的其他细胞因子相比，血清IL-27、IFNβ和IL-10的浓度相对较高。总之，从疾病发作开始每周三次给予JNJ-47965567不会改变SOD1小鼠的疾病进展或分子及细胞参数。
