Pluripotent stem cell models of Blau syndrome reveal an IFN-γ-dependent inflammatory response in macrophages
Pluripotent stem cell models of Blau syndrome reveal an IFN-γ-dependent inflammatory response in macrophages
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DOI：
10.1016/j.jaci.2017.04.013
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发表时间：
2018-01-01
影响因子：
14.2
通讯作者：
Saito, Megumu K.
中科院分区：
文献类型：
--
作者：
Takada, Sanami;Kambe, Naotomo;Saito, Megumu K.
背景：BLAU综合征，或早发性结节病，是一种与核苷酸结合寡聚化结构域2(NOD2)突变相关的青少年起病的系统性肉芽肿。BLAU综合征导致自身炎症的机制尚不清楚，目前尚无有效的针对BLAU综合征的特效治疗方法。目的：为了阐明BLAU综合征患者自身炎症的机制，我们试图在人类标本中阐明疾病相关突变NOD2与炎症反应的关系。使用CRISPR-Cas9系统校正与疾病相关的IPSC的NOD2突变，以准确评估IPSC来源的细胞的体外表型。我们还将相同的NOD2突变引入对照IPSC系。然后将这些同基因的IPSCs分化为单核细胞系，并观察核因子-kappaB途径和促炎细胞因子的分泌状态。结果：干扰素-γ通过上调NOD2起到启动信号的作用。在含有突变NOD2的IPSC来源的巨噬细胞中，干扰素-γ处理诱导非配体依赖的核因子-kappaB激活和促炎细胞因子的产生。RNA测序分析显示，突变的巨噬细胞在干扰素-γ治疗前后的转录图谱不同。患者来源的巨噬细胞表现出类似的干扰素-γ依赖的炎症反应。结论：我们的数据支持配体非依赖性自身炎症在BLAU综合征的病理生理学中的重要性。我们全面的等基因疾病特异性IPSC小组为探索BLAU综合征患者的治疗和诊断线索提供了一个有用的平台。
Background: Blau syndrome, or early-onset sarcoidosis, is a juvenile-onset systemic granulomatosis associated with a mutation in nucleotide-binding oligomerization domain 2 (NOD2). The underlying mechanisms of Blau syndrome leading to autoinflammation are still unclear, and there is currently no effective specific treatment for Blau syndrome.Objectives: To elucidate the mechanisms of autoinflammation in patients with Blau syndrome, we sought to clarify the relation between disease-associated mutant NOD2 and the inflammatory response in human samples.Methods: Blau syndrome specific induced pluripotent stem cell (iPSC) lines were established. The disease-associated NOD2 mutation of iPSCs was corrected by using a CRISPR-Cas9 system to precisely evaluate the in vitro phenotype of iPSC-derived cells. We also introduced the same NOD2 mutation into a control iPSC line. These isogenic iPSCs were then differentiated into monocytic cell lineages, and the statuses of nuclear factor kappa B pathway and proinflammatory cytokine secretion were investigated.Results: IFN-gamma acted as a priming signal through upregulation of NOD2. In iPSC-derived macrophages with mutant NOD2, IFN-gamma treatment induced ligand-independent nuclear factor kappa B activation and proinflammatory cytokine production. RNA sequencing analysis revealed distinct transcriptional profiles of mutant macrophages both before and after IFN-gamma treatment. Patient-derived macrophages demonstrated a similar IFN-gamma dependent inflammatory response.Conclusions: Our data support the significance of ligand-independent autoinflammation in the pathophysiology of Blau syndrome. Our comprehensive isogenic disease-specific iPSC panel provides a useful platform for probing therapeutic and diagnostic clues for the treatment of patients with Blau syndrome.