Effect of siRNA-mediated GSK3 knock-down on the transcriptional signature of human GM-CSF-dependent monocyte-derived macrophages

The functional versatility of macrophages is intrincately tied to factors such as their ontogeny and the specific tissue and extracellular environment. Monocyte-derived macrophages are oppositely instructed by M-CSF or GM-CSF. GM-CSF drives monocyte-derived macrophages towards heightened pro-inflammatory activity and the acquisition of the lung alveolar macrophage phenotype and gene profile whereas M-CSF gives rise to anti-inflammatory, pro-resolving, and immunosuppressive monocyte-derived macrophages. We explored the molecular impact of siRNA mediated knocking-down GSK3A, GSK3B or both (GSK3A/B) on the gene expression profile of GM-CSF-primed human monocyte derived macrophages. GSK3A/B knowdown skewed the transcriptional profile of GM-MØ towards an anti-inflammatory phenotype. mRNA profiles of human macrophages differentiated with GM-CSF (GM-MØ) and transfected with siRNA control (siCNT), siRNA for GSK3A (siGSK3A), siRNA for GSK3B (siGSK3B) or both (siGSK3A and siGSK3B) and analyzed 72 hours later.

巨噬细胞（macrophages）的功能多样性与其个体发育、所处特定组织及细胞外微环境等因素紧密相关。单核细胞来源的巨噬细胞的极化方向会因M-CSF与GM-CSF的刺激而呈现截然不同的调控结果：GM-CSF可促使单核细胞来源的巨噬细胞向促炎活性增强的方向分化，并获得肺泡巨噬细胞的表型与基因表达特征；而M-CSF则诱导产生具有抗炎、促炎症消退及免疫抑制特性的单核细胞来源巨噬细胞。本研究探究了小干扰RNA（small interfering RNA, siRNA）介导的GSK3A、GSK3B或二者联合（GSK3A/B）敲低，对经GM-CSF预处理的人单核细胞来源巨噬细胞基因表达谱的分子调控效应。实验结果表明，GSK3A/B敲低可将GM-MØ的转录谱向抗炎表型偏移。本实验的分析样本为经GM-CSF诱导分化的人巨噬细胞（GM-MØ），该类细胞分别被转染对照siRNA（siCNT）、靶向GSK3A的siRNA（siGSK3A）、靶向GSK3B的siRNA（siGSK3B）或二者联合转染（siGSK3A与siGSK3B），并在转染72小时后进行mRNA表达谱分析。