Retention of normal glia function by an isoform-selective protein kinase inhibitor drug candidate that modulates cytokine production and cognitive outcomes.
Retention of normal glia function by an isoform-selective protein kinase inhibitor drug candidate that modulates cytokine production and cognitive outcomes.
复制标题
DOI：
10.1186/s12974-017-0845-2
复制
发表时间：
2017-04-05
影响因子：
9.3
通讯作者：
Van Eldik LJ
中科院分区：
文献类型：
--
作者：
Zhou Z;Bachstetter AD;Späni CB;Roy SM;Watterson DM;Van Eldik LJ
脑内p38 α丝裂原活化蛋白激酶（MAPK）是一种基于病理生理学进展的神经炎症-突触功能障碍循环的潜在治疗靶点，通过抑制胶质细胞和神经元中相同的活化靶点，提供了一种创新的药理学策略，从而提高疗效的可能性。高选择性脑渗透性p38 α MAPK抑制剂MW150可减轻两种不同阿尔茨海默病（AD）相关模型中的认知功能障碍，并避免了先前混合激酶抑制剂候选药物遇到的问题。因此，至关重要的是，这种CNS活性激酶抑制剂的神经胶质细胞的影响得到解决，以预期未来的临床研究中使用。我们在AD相关APP/PS1基因敲入（KI）小鼠模型中探索了MW150对神经胶质生物学的影响，我们先前在该模型中显示了对抑制突触依赖性联想和空间记忆缺陷的疗效。每天向11 - 12月龄KI小鼠给予MW150（2.5 mg/kg/天），持续14天，并使用ELISA测量小鼠皮质匀浆中促炎细胞因子IL-1 β、TNF α和IL-6的水平。通过免疫组化评估胶质细胞标志物IBA1、CD45、CD68和GFAP。通过免疫荧光染色，然后通过共聚焦成像来定量小胶质细胞和淀粉样蛋白斑块。采用ELISA法测定可溶性和不溶性A β 40和A β 42的水平。对神经炎症标志物的体内药效学效应研究通过在小鼠小胶质细胞BV2细胞系中进行的机制研究进行补充，使用活细胞成像技术监测增殖、迁移和吞噬活性。在确定的治疗时间窗内对KI小鼠进行MW150干预可减弱IL-1 β和TNF α水平的升高，但对IL-6无影响。MW150处理还增加了淀粉样蛋白斑块15 μ m半径内的IBA1+小胶质细胞，而不显著影响总体小胶质细胞或斑块体积。MW150处理对IBA1、CD45、CD68、GFAP、A β 40和A β 42水平无影响。MW150没有显著改变BV2细胞中的小胶质细胞迁移、增殖或吞噬作用。我们的研究结果表明，有效剂量的MW150可以选择性地调节与病理进展相关的神经炎症反应，而不会对小胶质细胞的正常生理功能产生泛抑制。本文的在线版本（doi：10.1186/s12974 - 017 - 0845 - 2）包含补充材料，可供授权用户使用。
Brain p38α mitogen-activated protein kinase (MAPK), a potential therapeutic target for cognitive dysfunction based on the neuroinflammation-synaptic dysfunction cycle of pathophysiology progression, offers an innovative pharmacological strategy via inhibiting the same activated target in both glia and neurons, thereby enhancing the possibility for efficacy. The highly selective, brain-penetrant p38αMAPK inhibitor MW150 attenuates cognitive dysfunction in two distinct Alzheimer’s disease (AD)-relevant models and avoids the problems encountered with previous mixed-kinase inhibitor drug candidates. Therefore, it is essential that the glial effects of this CNS-active kinase inhibitor be addressed in order to anticipate future use in clinical investigations. We explored the effects of MW150 on glial biology in the AD-relevant APP/PS1 knock-in (KI) mouse model where we previously showed efficacy in suppression of hippocampal-dependent associative and spatial memory deficits. MW150 (2.5 mg/kg/day) was administered daily to 11–12-month-old KI mice for 14 days, and levels of proinflammatory cytokines IL-1β, TNFα, and IL-6 measured in homogenates of mouse cortex using ELISA. Glial markers IBA1, CD45, CD68, and GFAP were assessed by immunohistochemistry. Microglia and amyloid plaques were quantified by immunofluorescence staining followed by confocal imaging. Levels of soluble and insoluble of Aβ40 and Aβ42 were measured by ELISA. The studies of in vivo pharmacodynamic effects on markers of neuroinflammation were complemented by mechanistic studies in the murine microglia BV2 cell line, using live cell imaging techniques to monitor proliferation, migration, and phagocytosis activities. Intervention with MW150 in KI mice during the established therapeutic time window attenuated the increased levels of IL-1β and TNFα but not IL-6. MW150 treatment also increased the IBA1+ microglia within a 15 μm radius of the amyloid plaques, without significantly affecting overall microglia or plaque volume. Levels of IBA1, CD45, CD68, GFAP, and Aβ40 and Aβ42 were not affected by MW150 treatment. MW150 did not significantly alter microglial migration, proliferation, or phagocytosis in BV2 cells. Our results demonstrate that MW150 at an efficacious dose can selectively modulate neuroinflammatory responses associated with pathology progression without pan-suppression of normal physiological functions of microglia. The online version of this article (doi:10.1186/s12974-017-0845-2) contains supplementary material, which is available to authorized users.