Department of Pharmacology, Neuroscience and Mental Health Institute and Glycomics Institute of Alberta, University of Alberta, Edmonton, AB, Canada.
Department of Neurology, Johns Hopkins University School of Medicine, Baltimore, MD, USA.
J Neuroinflammation. 2023 Nov 23;20(1):276. doi: 10.1186/s12974-023-02963-y.
Chronic activation and dysfunction of microglia have been implicated in the pathogenesis and progression of many neurodegenerative disorders, including Huntington's disease (HD). HD is a genetic condition caused by a mutation that affects the folding and function of huntingtin (HTT). Signs of microglia activation have been observed in HD patients even before the onset of symptoms. It is unclear, however, whether pro-inflammatory microglia activation in HD results from cell-autonomous expression of mutant HTT, is the response of microglia to a diseased brain environment, or both. In this study, we used primary microglia isolated from HD knock-in (Q140) and wild-type (Q7) mice to investigate their response to inflammatory conditions in vitro in the absence of confounding effects arising from brain pathology. We show that naïve Q140 microglia do not undergo spontaneous pro-inflammatory activation and respond to inflammatory triggers, including stimulation of TLR4 and TLR2 and exposure to necrotic cells, with similar kinetics of pro-inflammatory gene expression as wild-type microglia. Upon termination of the inflammatory insult, the transcription of pro-inflammatory cytokines is tapered off in Q140 and wild-type microglia with similar kinetics. However, the ability of Q140 microglia to develop tolerance in response to repeated inflammatory stimulations is partially impaired in vitro and in vivo, potentially contributing to the establishment of chronic neuroinflammation in HD. We further show that ganglioside GM1, a glycosphingolipid with anti-inflammatory effects on wild-type microglia, not only decreases the production of pro-inflammatory cytokines and nitric oxide in activated Q140 microglia, but also dramatically dampen microglia response to re-stimulation with LPS in an experimental model of tolerance. These effects are independent from the expression of interleukin 1 receptor associated kinase 3 (Irak-3), a strong modulator of LPS signaling involved in the development of innate immune tolerance and previously shown to be upregulated by immune cell treatment with gangliosides. Altogether, our data suggest that external triggers are required for HD microglia activation, but a cell-autonomous dysfunction that affects the ability of HD microglia to acquire tolerance might contribute to the establishment of neuroinflammation in HD. Administration of GM1 might be beneficial to attenuate chronic microglia activation and neuroinflammation.
慢性小胶质细胞激活和功能障碍与许多神经退行性疾病的发病机制和进展有关,包括亨廷顿病 (HD)。HD 是一种由影响亨廷顿蛋白 (HTT) 折叠和功能的突变引起的遗传疾病。即使在症状出现之前,HD 患者也已经观察到小胶质细胞激活的迹象。然而,HD 中小胶质细胞的促炎激活是源自突变 HTT 的细胞自主表达,还是对患病大脑环境的反应,或者两者兼而有之,目前尚不清楚。在这项研究中,我们使用从小鼠 HD 基因敲入 (Q140) 和野生型 (Q7) 中分离的原代小胶质细胞,在不存在脑病理学引起的混杂效应的情况下,研究它们在体外对炎症条件的反应。我们表明,幼稚的 Q140 小胶质细胞不会自发发生促炎激活,并且对炎症触发因素(包括 TLR4 和 TLR2 的刺激以及暴露于坏死细胞)的反应与野生型小胶质细胞相似,其促炎基因表达的动力学相似。在炎症刺激终止后,Q140 和野生型小胶质细胞中的促炎细胞因子转录以相似的动力学减弱。然而,Q140 小胶质细胞在体外和体内对反复炎症刺激产生耐受的能力部分受损,这可能导致 HD 中的慢性神经炎症的建立。我们进一步表明,神经节苷脂 GM1 是一种具有抗炎作用的糖脂,不仅可以降低激活的 Q140 小胶质细胞中促炎细胞因子和一氧化氮的产生,而且可以显著抑制 LPS 在耐受实验模型中小胶质细胞对 LPS 的再刺激反应。这些作用独立于白细胞介素 1 受体相关激酶 3 (Irak-3) 的表达,后者是 LPS 信号转导的一个强有力调节剂,参与先天免疫耐受的发展,先前已被证明通过神经节苷脂处理免疫细胞而被上调。总的来说,我们的数据表明,外部触发因素是 HD 小胶质细胞激活所必需的,但一种影响 HD 小胶质细胞获得耐受能力的自主功能障碍可能有助于 HD 中神经炎症的建立。GM1 的给药可能有益于减轻慢性小胶质细胞激活和神经炎症。