Direct conversion of human fibroblasts into functional osteoblasts by defined factors
Direct conversion of human fibroblasts into functional osteoblasts by defined factors
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DOI：
10.1073/pnas.1420713112
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发表时间：
2015-05-12
影响因子：
11.1
通讯作者：
Mazda, Osam
中科院分区：
文献类型：
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作者：
Yamamoto, Kenta;Kishida, Tsunao;Mazda, Osam
成骨细胞产生钙化骨基质并有助于骨形成和重塑。在这项研究中，我们建立了一种通过转导一些确定的因子并在成骨培养基中培养来直接将人成纤维细胞转化为成骨细胞的程序。成骨细胞特异性转录因子、Runt 相关转录因子 2 (Runx2) 和 Osterix，与八聚体结合转录因子 3/4 (Oct4) 和 L-Myc (RXOL) 转导相结合，将大约 80% 的成纤维细胞转化为产生骨钙素的细胞。 RXOL诱导的直接转化成骨细胞（dOB）显示出与正常人成骨细胞相似的基因表达谱，并在移植到人工骨缺损病变处的免疫缺陷小鼠体内后有助于骨修复。 dOB 表达内源 Runx2 和 Osterix，并且不需要外源基因的持续表达来维持其表型。另一种组合，Oct4 加 L-Myc (OL)，也诱导成纤维细胞产生骨基质，但 OL 转导的细胞不表达 Osterix，并且与 RXOL 转导的细胞相比，与成骨细胞表现出更远的基因表达谱。这些发现强烈表明，RXOL 可以成功地将成纤维细胞直接重编程为功能性成骨细胞，这项技术可以提供针对骨疾病的骨再生治疗。
Osteoblasts produce calcified bone matrix and contribute to bone formation and remodeling. In this study, we established a procedure to directly convert human fibroblasts into osteoblasts by transducing some defined factors and culturing in osteogenic medium. Osteoblast-specific transcription factors, Runt-related transcription factor 2 (Runx2), and Osterix, in combination with Octamer-binding transcription factor 3/4 (Oct4) and L-Myc (RXOL) transduction, converted similar to 80% of the fibroblasts into osteocalcin-producing cells. The directly converted osteoblasts (dOBs) induced by RXOL displayed a similar gene expression profile as normal human osteoblasts and contributed to bone repair after transplantation into immunodeficient mice at artificial bone defect lesions. The dOBs expressed endogenous Runx2 and Osterix, and did not require continuous expression of the exogenous genes to maintain their phenotype. Another combination, Oct4 plus L-Myc (OL), also induced fibroblasts to produce bone matrix, but the OL-transduced cells did not express Osterix and exhibited a more distant gene expression profile to osteoblasts compared with RXOL-transduced cells. These findings strongly suggest successful direct reprogramming of fibroblasts into functional osteoblasts by RXOL, a technology that may provide bone regeneration therapy against bone disorders.