In response to inflammatory cytokines and chemokines, monocytes differentiate into macrophages. Comprehensive analysis of gene expression regulation of neuronal guidance cue (NGC) ligands and receptors in the monocyte-to-macrophage differentiation process is not available yet. We performed transcriptome profiling in both human primary PBMCs/PBMC-derived macrophages and THP-1 cells/THP-1-macrophages using microarray or RNA sequencing methods. Pathway analysis showed that the axonal guidance pathway is significantly regulated upon monocyte differentiation. We confirmed NGC ligands and receptors which were consistently regulated, including SEMA4D, SEMA7A, NRP1, NRP2, PLXNA1 and PLXNA3. The involvement of RNA-binding protein quaking (QKI) in the regulation of NGC expression was investigated using monocytes and macrophages from a QKI haplo-insufficient patient and her healthy sibling. This revealed a positive correlation of SEMA7A expression with QKI expression. In silico analysis of 3′UTRs of NGCs proposed the competitive binding of QKI to proximal microRNA targeting sites as the mechanism of QKI-dependent regulation of SEMA7A. RNA immunoprecipitation confirmed an interaction of QKI with the 3′UTR of SEMA7A. Loss of SEMA7A resulted in monocyte differentiation towards a more anti-inflammatory macrophage. Taken together, the axonal guidance pathway is regulated during monocyte-to-macrophage differentiation, and the regulation is in line with the necessary functional adaption for the specialised role of macrophages.

响应炎性细胞因子和趋化因子，单核细胞分化成巨噬细胞。尚未全面分析单核细胞-巨噬细胞分化过程中神经元引导线索 (NGC) 配体和受体的基因表达调控。我们使用微阵列或 RNA 测序方法对人类原代 PBMC/PBMC 衍生的巨噬细胞和 THP-1 细胞/THP-1-巨噬细胞进行了转录组分析。通路分析表明，轴突引导通路在单核细胞分化时受到显着调节。我们确认了受到一致调节的 NGC 配体和受体，包括 SEMA4D、SEMA7A、NRP1、NRP2、PLXNA1 和 PLXNA3。使用来自 QKI 单倍体不足的患者及其健康兄弟姐妹的单核细胞和巨噬细胞研究了 RNA 结合蛋白 quaking (QKI) 在 NGC 表达调节中的作用。这揭示了 SEMA7A 表达与 QKI 表达的正相关。NGC 的 3'UTR 的计算机分析提出 QKI 与近端 microRNA 靶向位点的竞争性结合是 QKI 依赖性 SEMA7A 调节的机制。RNA 免疫沉淀证实了 QKI 与 SEMA7A 的 3'UTR 的相互作用。SEMA7A 的缺失导致单核细胞分化为更具抗炎性的巨噬细胞。总之，轴突引导通路在单核细胞向巨噬细胞分化过程中受到调节，这种调节与巨噬细胞特殊作用所必需的功能适应相一致。