TAT-mediated neurogenin 3 protein transduction stimulates pancreatic endocrine differentiation in vitro
TAT-mediated neurogenin 3 protein transduction stimulates pancreatic endocrine differentiation in vitro
复制标题
DOI：
10.2337/diabetes.54.3.720
复制
发表时间：
2005-03-01
期刊：
影响因子：
7.7
通讯作者：
Edlund, H
中科院分区：
文献类型：
--
作者：
Domínguez-Bendala, J;Klein, D;Edlund, H
干细胞技术在治疗 I 型糖尿病方面具有巨大潜力，只要能够以有效的方式选择性地产生功能性可移植 R 细胞。这样的过程至少在一定程度上应该重现β细胞的体外胚胎发育。然而，在鉴定参与 P 细胞发育的转录因子方面取得的进展并没有伴随着阐明其指导性细胞外信号模式的成功。在这里，我们提出了一种基于 HIV/TAT 蛋白转导结构域的使用的新方法的原理证明，以规避这个问题。神经原素 3 (ngn3) 是一种其表达对于胰腺内分泌分化至关重要的因子，与 TAT 结构域融合。对培养的胰腺外植体施用 TAT/ngn3 会导致下游报告基因和内源基因的有效摄取、核转位和刺激。与蛋白质的预测活性一致，在 TAT/ngn3 存在下培养的 e9.5 和 e13.5 小鼠胰腺外植体显示与对照样品相比内分泌分化水平增加。我们的结果提出了通过使用适当的序列和 TAT 融合转录因子的组合，将干细胞/祖细胞顺序指定为 P 细胞谱系的可能性。
Stem cell technologies hold great potential for the treatment of type I diabetes, provided that functional transplantable R-cells can be selectively generated in an efficient manner. Such a process should recapitulate, at least to a certain extent, the embryonic development of beta-cells in vitro. However, progress at identifying the transcription factors involved in P-cell development has not been accompanied by a parallel success at unraveling the pattern of their instructive extracellular signals. Here we present proof of principle of a novel approach to circumvent this problem, based on the use of the HIV/TAT protein transduction domain. Neurogenin 3 (ngn3), a factor whose expression is essential for Pancreatic endocrine differentiation, was fused to the TAT domain. Administration of TAT/ngn3 to cultured pancreatic explants results in efficient uptake, nuclear translocation, and stimulation of downstream reporter and endogenous genes. Consistent with the predicted activity of the protein, e9.5 and e13.5 mouse pancreatic explants cultured in the presence of TAT/ngn3 show an increased level of endocrine differentiation compared with control samples. Our results raise the possibility of sequentially specifying stem/progenitor cells toward the P-cell lineage, by using the appropriate sequence and combination of TAT-fused transcription factors.