Human iPS cell-derived mural cells as an in vitro model of hereditary cerebral small vessel disease
Human iPS cell-derived mural cells as an in vitro model of hereditary cerebral small vessel disease
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DOI：
10.1186/s13041-020-00573-w
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发表时间：
2020-03-19
期刊：
影响因子：
3.6
通讯作者：
Inoue, Haruhisa
中科院分区：
文献类型：
--
作者：
Yamamoto, Yumi;Kojima, Katsutoshi;Inoue, Haruhisa
伴有皮质下梗死和白质脑病的常染色体显性遗传性脑动脉病（CADASIL）是遗传性脑小血管疾病的最常见形式之一，由NOTCH 3突变引起。我们的研究小组以前曾报道过在CADASIL特异性颗粒嗜锇物质中掺入NOTCH 3细胞外结构域（N3 ECD），以及在CADASIL死后大脑中增加PDGFR β免疫反应性。在这里，我们的目的是建立一个体外模型的CADASIL，它可以概括这些CADASIL表型，使用诱导多能干细胞（iPSC）。我们已经完善了内皮细胞的分化协议，以获得成熟的壁细胞（MCs）与他们的特性。将来自3名携带p.Arg182Cys、p.Arg141Cys和p.Cys106Arg突变的CADASIL患者的iPSC分化为MC，并比较其功能和分子特征。分化的CADASIL MCs重现了先前报道的致病性变化：PDGFR β增加和丝状肌动蛋白网络的异常结构/分布，以及N3 ECD/LTBP-1/HtrA 1免疫阳性沉积物。CADASIL MCs的迁移率增加，但受到NOTCH 3或PDGFRB敲低的抑制。CADASIL MCs显示对PDGF-BB的反应性改变。患者来源的MC可以概括CADASIL病理，因此有助于了解发病机制和制定潜在的治疗策略。
Cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy (CADASIL) is one of the most common forms of hereditary cerebral small vessel diseases and is caused by mutations in NOTCH3. Our group has previously reported incorporation of NOTCH3 extracellular domain (N3ECD) in the CADASIL-specific granular osmiophilic materials and increase of PDGFR beta immunoreactivity in CADASIL postmortem brains. Here, we aimed to establish an in vitro model of CADASIL, which can recapitulate those CADASIL phenotypes, using induced pluripotent stem cells (iPSCs). We have refined a differentiation protocol of endothelial cells to obtain mature mural cells (MCs) with their characteristic properties. iPSCs from three CADASIL patients with p.Arg182Cys, p.Arg141Cys and p.Cys106Arg mutations were differentiated into MCs and their functional and molecular profiles were compared. The differentiated CADASIL MCs recapitulated pathogenic changes reported previously: increased PDGFR beta and abnormal structure/distribution of filamentous actin network, as well as N3ECD/LTBP-1/HtrA1-immunopositive deposits. Migration rate of CADASIL MCs was enhanced but suppressed by knockdown of NOTCH3 or PDGFRB. CADASIL MCs showed altered reactivity to PDGF-BB. Patient-derived MCs can recapitulate CADASIL pathology and are therefore useful in understanding the pathogenesis and developing potential treatment strategies.