Zhang Hanrui, Xue Chenyi, Shah Rhia, Bermingham Kate, Hinkle Christine C, Li Wenjun, Rodrigues Amrith, Tabita-Martinez Jennifer, Millar John S, Cuchel Marina, Pashos Evanthia E, Liu Ying, Yan Ruilan, Yang Wenli, Gosai Sager J, VanDorn Daniel, Chou Stella T, Gregory Brian D, Morrisey Edward E, Li Mingyao, Rader Daniel J, Reilly Muredach P
From the Cardiovascular Institute (H.Z., C.X., R.S., K.B., C.C.H., W.L., A.R., J.T.-M., E.E.P., E.E.M., D.J.R., M.P.R.), and Department of Biostatistics and Epidemiology (M.L.), Perelman School of Medicine, Institute for Translational Medicine and Therapeutics, Institute for Diabetes, Obesity, and Metabolism (A.R., M.C., E.E.P., D.J.R.), Department of Medicine, Metabolic Tracer Resource, Institute for Diabetes, Obesity, and Metabolism (J.S.M.), Institute for Regenerative Medicine (Y.L., R.Y., W.Y., E.E.M.), Department of Biology, Perelman School of Medicine and School of Arts and Science (S.J.G., B.D.G.), PENN Genome Frontiers Institute (S.J.G., B.D.G.), Department of Pediatrics, Perelman School of Medicine (S.T.C.), Department of Cell and Developmental Biology, Perelman School of Medicine (E.E.M.), and Department of Biostatistics and Epidemiology, Perelman School of Medicine, University of Pennsylvania, Philadelphia; and Division of Hematology, The Children's Hospital of Philadelphia, PA (D.V., S.T.C.).
Circ Res. 2015 Jun 19;117(1):17-28. doi: 10.1161/CIRCRESAHA.117.305860. Epub 2015 Apr 22.
An efficient and reproducible source of genotype-specific human macrophages is essential for study of human macrophage biology and related diseases.
To perform integrated functional and transcriptome analyses of human induced pluripotent stem cell-derived macrophages (IPSDMs) and their isogenic human peripheral blood mononuclear cell-derived macrophage (HMDM) counterparts and assess the application of IPSDM in modeling macrophage polarization and Mendelian disease.
We developed an efficient protocol for differentiation of IPSDM, which expressed macrophage-specific markers and took up modified lipoproteins in a similar manner to HMDM. Like HMDM, IPSDM revealed reduction in phagocytosis, increase in cholesterol efflux capacity and characteristic secretion of inflammatory cytokines in response to M1 (lipopolysaccharide+interferon-γ) activation. RNA-Seq revealed that nonpolarized (M0) as well as M1 or M2 (interleukin-4) polarized IPSDM shared transcriptomic profiles with their isogenic HMDM counterparts while also revealing novel markers of macrophage polarization. Relative to IPSDM and HMDM of control individuals, patterns of defective cholesterol efflux to apolipoprotein A-I and high-density lipoprotein-3 were qualitatively and quantitatively similar in IPSDM and HMDM of patients with Tangier disease, an autosomal recessive disorder because of mutations in ATP-binding cassette transporter AI. Tangier disease-IPSDM also revealed novel defects of enhanced proinflammatory response to lipopolysaccharide stimulus.
Our protocol-derived IPSDM are comparable with HMDM at phenotypic, functional, and transcriptomic levels. Tangier disease-IPSDM recapitulated hallmark features observed in HMDM and revealed novel inflammatory phenotypes. IPSDMs provide a powerful tool for study of macrophage-specific function in human genetic disorders as well as molecular studies of human macrophage activation and polarization.
对于人类巨噬细胞生物学及相关疾病的研究而言，高效且可重复的基因型特异性人类巨噬细胞来源至关重要。
对人类诱导多能干细胞衍生的巨噬细胞（IPSDM）及其同基因的人类外周血单个核细胞衍生的巨噬细胞（HMDM）进行综合功能和转录组分析，并评估IPSDM在巨噬细胞极化和孟德尔疾病建模中的应用。
我们开发了一种高效的IPSDM分化方案，其表达巨噬细胞特异性标志物，并以与HMDM相似的方式摄取修饰的脂蛋白。与HMDM一样，IPSDM在M1（脂多糖+干扰素-γ）激活后，吞噬作用降低，胆固醇流出能力增加，且有特征性的炎性细胞因子分泌。RNA测序显示，非极化（M0）以及M1或M2（白细胞介素-4）极化的IPSDM与其同基因的HMDM具有相同的转录组图谱，同时也揭示了巨噬细胞极化的新标志物。与对照个体的IPSDM和HMDM相比，患有常染色体隐性疾病丹吉尔病（因ATP结合盒转运体AI突变所致）的患者的IPSDM和HMDM中，向载脂蛋白A-I和高密度脂蛋白-3的胆固醇流出缺陷模式在定性和定量上相似。丹吉尔病-IPSDM还揭示了对脂多糖刺激的促炎反应增强的新缺陷。
我们方案衍生的IPSDM在表型、功能和转录组水平上与HMDM相当。丹吉尔病-IPSDM重现了在HMDM中观察到的标志性特征，并揭示了新的炎症表型。IPSDM为研究人类遗传疾病中巨噬细胞特异性功能以及人类巨噬细胞激活和极化的分子研究提供了有力工具。
