Department of Oral Health and Diagnostic Sciences, School of Dental Medicine, University of Connecticut Health, 263 Farmington Avenue, Farmington, CT 06030, USA.
Department of Oral Health and Diagnostic Sciences, School of Dental Medicine, University of Connecticut Health, 263 Farmington Avenue, Farmington, CT 06030, USA.
Stem Cell Reports. 2017 Nov 14;9(5):1369-1376. doi: 10.1016/j.stemcr.2017.09.016. Epub 2017 Oct 19.
We identified osteoclast defects in craniometaphyseal dysplasia (CMD) using an easy-to-use protocol for differentiating osteoclasts from human induced pluripotent stem cells (hiPSCs). CMD is a rare genetic bone disorder, characterized by life-long progressive thickening of craniofacial bones and abnormal shape of long bones. hiPSCs from CMD patients with an in-frame deletion of Phe377 or Ser375 in ANKH are more refractory to in vitro osteoclast differentiation than control hiPSCs. To exclude differentiation effects due to genetic variability, we generated isogenic hiPSCs, which have identical genetic background except for the ANKH mutation. Isogenic hiPSCs with ANKH mutations formed fewer osteoclasts, resorbed less bone, expressed lower levels of osteoclast marker genes, and showed decreased protein levels of ANKH and vacuolar proton pump v-ATP6v0d2. This proof-of-concept study demonstrates that efficient and reproducible differentiation of isogenic hiPSCs into osteoclasts is possible and a promising tool for investigating mechanisms of CMD or other osteoclast-related disorders.
我们使用一种易于使用的方案从人诱导多能干细胞（hiPSC）中区分破骨细胞，从而鉴定出颅颌面骨发育不良（CMD）中的破骨细胞缺陷。CMD 是一种罕见的遗传性骨疾病，其特征是终生进行性颅面骨增厚和长骨形状异常。ANKH 中 Phe377 或 Ser375 框内缺失的 CMD 患者的 hiPSC 比对照 hiPSC 更难以进行体外破骨细胞分化。为了排除由于遗传变异引起的分化效应，我们生成了同基因 hiPSC，它们除了 ANKH 突变外具有相同的遗传背景。具有 ANKH 突变的同基因 hiPSC 形成的破骨细胞较少，吸收的骨量较少，表达的破骨细胞标记基因水平较低，并且 ANKH 和液泡质子泵 v-ATP6v0d2 的蛋白水平降低。这项概念验证研究表明，将同基因 hiPSC 有效地和可再现地分化为破骨细胞是可能的，并且是研究 CMD 或其他破骨细胞相关疾病的机制的有前途的工具。
