School of Biological Sciences, Nanyang Technological University, 637551 Singapore.
Nanyang Institute of Technology in Health and Medicine, Interdisciplinary Graduate School (IGS), Nanyang Technological University, 637551 Singapore.
Nucleic Acids Res. 2018 Jul 6;46(12):6069-6086. doi: 10.1093/nar/gky401.
We report the detailed transcriptomic profiles of human innate myeloid cells using RNA sequencing. Monocytes migrate from blood into infected or wounded tissue to differentiate into macrophages, and control inflammation via phagocytosis or cytokine secretion. We differentiated culture primary monocytes with either GM- or M-CSF to obtain pro- or anti-inflammatory macrophages, and respectively activated them with either LPS/IFNγ or anti-inflammatory cytokines. We also treated the THP-1 monocytic cell line with PMA and similar cytokines to mimic differentiation and activation. We detected thousands of expression and alternative-splicing changes during monocyte-to-macrophage differentiation and activation, and a net increase in exon inclusion. MBNL1 knockdown phenocopies several alternative-splicing changes and strongly impairs PMA differentiation, suggesting functional defects in monocytes from Myotonic Dystrophy patients. This study provides general insights into alternative splicing in the monocyte-macrophage lineage, whose future characterization will elucidate their contribution to immune functions, which are altered in immunodeficiencies, autoimmunity, atherosclerosis and cancer.
我们使用 RNA 测序报告了人类固有髓样细胞的详细转录组谱。单核细胞从血液迁移到感染或受伤的组织中，分化为巨噬细胞，并通过吞噬作用或细胞因子分泌来控制炎症。我们用 GM-CSF 或 M-CSF 分化培养原代单核细胞，获得促炎或抗炎巨噬细胞，并分别用 LPS/IFNγ 或抗炎细胞因子激活它们。我们还用 PMA 和类似细胞因子处理 THP-1 单核细胞系，以模拟分化和激活。我们在单核细胞向巨噬细胞分化和激活过程中检测到数千个表达和可变剪接变化，外显子包含增加。MBNL1 敲低模拟了几种可变剪接变化，并严重损害了 PMA 的分化，表明肌强直性营养不良患者单核细胞存在功能缺陷。这项研究为单核细胞-巨噬细胞谱系中的可变剪接提供了一般性的见解，其未来的特征将阐明它们对免疫功能的贡献，这些功能在免疫缺陷、自身免疫、动脉粥样硬化和癌症中发生改变。
