Identification of a potent combination of osteogenic genes for bone regeneration using embryonic stem (ES) cell-based sensor
Identification of a potent combination of osteogenic genes for bone regeneration using embryonic stem (ES) cell-based sensor
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DOI：
10.1096/fj.06-7571com
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发表时间：
2007-06-01
期刊：
影响因子：
4.8
通讯作者：
Chung, Ung-il
中科院分区：
文献类型：
--
作者：
Ohba, Shinsuke;Ikeda, Toshiyuki;Chung, Ung-il
通过全面筛选来确定体内组织再生的有效生物活性因子仍然是再生医学的挑战。在这里，我们报告的开发的ES细胞为基础的监测系统，成骨分化，成骨基因的有效组合，使用这样的系统的识别，并评估其治疗潜力。从携带由大鼠I型胶原α（1）启动子的2.3kb片段驱动的表达GFP的转基因的小鼠中分离ES细胞。使用这些经过改造以在成骨分化中发出荧光的细胞，我们筛选了cDNA文库和主要成骨相关基因的组合。其中，组成型激活素受体样激酶6（caALK 6）和runt相关转录因子2（Runx 2）的组合是诱导荧光的最小单位。该组合有效地诱导各种细胞类型的成骨分化，包括终末分化的非成骨细胞。该组合的协同作用通过核心结合因子β（Cbfb）的蛋白质稳定、Runx 2-Cbfb复合物形成的诱导及其DNA结合而发生。此外，在小鼠颅骨骨缺损中，移植用组合物转导的成纤维细胞单层片在4周内实现了骨再生。因此，我们成功地确定了骨再生基因的有效组合，这有助于扩大细胞来源。Ohba，S.，Ikeda，T.，Kugimiya，F.，Yano，F.，Lichtler，A. C.的方法，中村，K.，Takato，T.，川口，H.，Chung，U. I.使用基于胚胎干细胞的传感器鉴定用于骨再生的成骨基因的有效组合。
To identify potent bioactive factors for in vivo tissue regeneration by comprehensive screening remains a challenge for regenerative medicine. Here we report the development of an ES cell-based monitoring system for osteogenic differentiation, the identification of a potent combination of osteogenic genes using such a system, and an evaluation of its therapeutic potentials. ES cells were isolated from mice carrying a transgene expressing GFP driven by the 2.3 kb fragment of rat type I collagen alpha( 1) promoter. Using these cells engineered to fluoresce on osteogenic differentiation, we screened cDNA libraries and combinations of major osteogenesis-related genes. Among them, the combination of constitutively active activin receptorlike kinase 6 (caALK6) and runt-related transcription factor 2 ( Runx2) was the minimal unit that induced fluorescence. The combination efficiently induced osteogenic differentiation in various cell types, including terminally differentiated nonosteogenic cells. The cooperative action of the combination occurred through protein stabilization of core binding factor beta (Cbfb), induction of Runx2-Cbfb complex formation, and its DNA binding. Furthermore, transplantation of a monolayer sheet of fibroblasts transduced with the combination achieved bone regeneration within 4 wk in mouse calvarial bone defects. Thus, we successfully identified the potent combination of genes for bone regeneration, which helped broaden cell sources.-Ohba, S., Ikeda, T., Kugimiya, F., Yano, F., Lichtler, A. C., Nakamura, K., Takato, T., Kawaguchi, H., Chung, U. I. Identification of a potent combination of osteogenic genes for bone regeneration using embryonic stem ( ES) cell-based sensor.