Mbd3 Promotes Reprogramming of Primary Human Fibroblasts.
Mbd3 Promotes Reprogramming of Primary Human Fibroblasts.
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DOI：
10.15283/ijsc18036
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发表时间：
2018-11-30
影响因子：
2.3
通讯作者：
Nathwani AC
中科院分区：
文献类型：
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作者：
Jaffer S;Goh P;Abbasian M;Nathwani AC
Mbd3（甲基- cpg结合域蛋白）是NuRD（核小体重构和去乙酰化）的核心成员，对胚胎发生至关重要。然而，其在体细胞重编程成诱导多能干细胞（iPSC）中的作用仍存在争议。一些报道表明Mbd3抑制多能性，而另一些则表明它大大提高了重编程效率。我们的研究首次评估了Mbd3在无饲料条件下使用Yamanaka episomal质粒（重编程因子（reprogramming factors， RF））对原代人成纤维细胞重编程的作用。我们发现shrna介导的Mbd3的部分缺失导致原代人成纤维细胞重编程效率降低了5倍。此外，敲除Mbd3后产生的iPSC即使表达了所有多能性标记，也不能进行三龄分化。相比之下，Mbd3b亚型和Yamanaka episomal质粒的过表达使成纤维细胞衍生的iPSC菌落的数量增加了至少两倍。所得菌落具有三龄分化能力。因此，我们的研究结果表明Mbd3似乎在原代人成纤维细胞的重编程中发挥了重要作用，这为重编程的生物学提供了进一步的见解，同时也对iPSC向临床的转化具有直接意义。
Mbd3 (Methyl-CpG binding domain protein), a core member of NuRD (nucleosome remodelling and deacetylation) is essential for embryogenesis. However, its role in reprogramming of somatic cells into induced pluripotent stem cells (iPSC) remains controversial. Some reports suggest that Mbd3 inhibits pluripotency, whilst others show that it greatly enhances reprogramming efficiency. Our study is the first to assess the role of Mbd3 on reprogramming of primary human fibroblasts using Yamanaka episomal plasmids (Reprogramming factors (RF) under feeder-free conditions. We showed that shRNA-mediated partial depletion of Mbd3 resulted in >5-fold reduction in the efficiency of reprogramming of primary human fibroblasts. Furthermore, iPSC that emerged after knock-down of Mbd3 were incapable of trilineage differentiation even though they expressed all markers of pluripotency. In contrast, over-expression of the Mbd3b isoform along with the Yamanaka episomal plasmids increased the number of fibroblast derived iPSC colonies by at least two-fold. The resulting colonies were capable of trilineage differentiation. Our results, therefore, suggest that Mbd3 appears to play an important role in reprogramming of primary human fibroblasts, which provides further insight into the biology of reprogramming but also has direct implication for translation of iPSC to clinic.