Department of Stem Cell Biology, Institute of Molecular Embryology and Genetics, Kumamoto University, Kumamoto, Japan.
1] Department of Stem Cell Biology, Institute of Molecular Embryology and Genetics, Kumamoto University, Kumamoto, Japan. [2] Department of Pediatrics, Graduate School of Medical Sciences, Kumamoto University, Kumamoto, Japan.
Nat Chem Biol. 2014 Feb;10(2):141-8. doi: 10.1038/nchembio.1410. Epub 2013 Dec 15.
Cell replacement therapy for diabetes mellitus requires cost-effective generation of high-quality, insulin-producing, pancreatic β cells from pluripotent stem cells. Development of this technique has been hampered by a lack of knowledge of the molecular mechanisms underlying β-cell differentiation. The present study identified reserpine and tetrabenazine (TBZ), both vesicular monoamine transporter 2 (VMAT2) inhibitors, as promoters of late-stage differentiation of Pdx1-positive pancreatic progenitor cells into Neurog3 (referred to henceforth as Ngn3)-positive endocrine precursors. VMAT2-controlled monoamines, such as dopamine, histamine and serotonin, negatively regulated β-cell differentiation. Reserpine or TBZ acted additively with dibutyryl adenosine 3',5'-cyclic AMP, a cell-permeable cAMP analog, to potentiate differentiation of embryonic stem (ES) cells into β cells that exhibited glucose-stimulated insulin secretion. When ES cell-derived β cells were transplanted into AKITA diabetic mice, the cells reversed hyperglycemia. Our protocol provides a basis for the understanding of β-cell differentiation and its application to a cost-effective production of functional β cells for cell therapy.
用于治疗糖尿病的细胞替代疗法需要从多能干细胞高效地生成高质量、可产生胰岛素的胰腺β细胞。由于缺乏对β细胞分化的分子机制的了解，该技术的发展一直受到阻碍。本研究发现，囊泡单胺转运体 2（VMAT2）抑制剂利血平和四苯嗪（TBZ）均可促进 Pdx1 阳性胰腺祖细胞向神经调节素 3（下文简称 Ngn3）阳性内分泌前体细胞的晚期分化。由 VMAT2 控制的单胺类物质，如多巴胺、组胺和血清素，负调控β细胞分化。利血平或 TBZ 与可渗透细胞的 cAMP 类似物二丁酰环磷腺苷（db-cAMP）协同作用，增强胚胎干细胞（ES 细胞）向可进行葡萄糖刺激胰岛素分泌的β细胞的分化。当 ES 细胞衍生的β细胞被移植到 AKITA 糖尿病小鼠中时，这些细胞逆转了高血糖症。我们的方案为理解β细胞分化及其在高效生产用于细胞治疗的功能性β细胞方面的应用提供了基础。
