Redifferentiation of expanded human islet β cells by inhibition of ARX.
Redifferentiation of expanded human islet β cells by inhibition of ARX.
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DOI：
10.1038/srep20698
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发表时间：
2016-02-09
影响因子：
4.6
通讯作者：
Efrat S
中科院分区：
文献类型：
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作者：
Friedman-Mazursky O;Elkon R;Efrat S
已经评价了成人胰岛β细胞的离体扩增以产生用于移植的丰富的胰岛素产生细胞;然而，谱系追踪已经证明该过程导致β细胞去分化。β-细胞衍生（BCD）细胞的再分化可以使用称为再分化混合物（RC）的可溶性因子的组合来实现;然而，这种处理仅导致一部分BCD细胞的再分化。本研究旨在通过影响RC处理激活的胰岛祖细胞转录因子的平衡来提高再分化效率。具体而言，RC处理诱导转录因子PAX 4和ARX，其分别在引导胰腺内分泌祖细胞进入β/δ或α/PP发育途径中起关键作用。在RC处理的BCD细胞中，ARX的误激活可能抑制其再分化为β细胞。通过shRNA阻断ARX表达，胰岛素mRNA水平升高了12.8倍，胰岛素阳性BCD细胞的数量增加了一倍多。在用RC处理的扩增的α-细胞衍生的细胞中，ARX抑制不会导致它们转分化为胰岛素产生细胞。RC和ARX shRNA联合治疗可能有助于产生大量的胰岛素产生细胞，用于移植到1型糖尿病患者体内。
Ex-vivo expansion of adult human islet β cells has been evaluated for generation of abundant insulin-producing cells for transplantation; however, lineage-tracing has demonstrated that this process results in β-cell dedifferentiation. Redifferentiation of β-cell-derived (BCD) cells can be achieved using a combination of soluble factors termed Redifferentiation Cocktail (RC); however, this treatment leads to redifferentiation of only a fraction of BCD cells. This study aimed at improving redifferentiation efficiency by affecting the balance of islet progenitor-cell transcription factors activated by RC treatment. Specifically, RC treatment induces the transcription factors PAX4 and ARX, which play key roles in directing pancreas endocrine progenitor cells into the β/δ or α/PP developmental pathways, respectively. Misactivation of ARX in RC-treated BCD cells may inhibit their redifferentiation into β cells. Blocking ARX expression by shRNA elevated insulin mRNA levels 12.8-fold, and more than doubled the number of insulin-positive BCD cells. ARX inhibition in expanded α-cell-derived cells treated with RC did not cause their transdifferentiation into insulin-producing cells. The combination of RC and ARX shRNA treatment may facilitate the generation of abundant insulin-producing cells for transplantation into patients with type 1 diabetes.