Rapid differentiation of hiPSCs into functional oligodendrocytes using an OLIG2 synthetic modified messenger RNA.
Rapid differentiation of hiPSCs into functional oligodendrocytes using an OLIG2 synthetic modified messenger RNA.
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DOI：
10.1038/s42003-022-04043-y
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发表时间：
2022-10-14
影响因子：
5.9
通讯作者：
中科院分区：
文献类型：
--
作者：
已经引入转录因子（TF）来驱动人诱导的多能干细胞（hiPSC）高效分化为谱系特异性少突胶质细胞（OL）。然而，有效的策略目前主要依赖于基因组整合病毒。在这里，我们展示了一种基于合成修饰信使RNA（smRNA）的重编程方法，该方法可以产生无转基因的OL。编码修饰形式的0 LIG 2的smRNA（其中丝氨酸147磷酸化位点被丙氨酸取代，0 LIG 2S 147 A）被设计用于将hiPSC重编程为OL。我们证明，重复施用编码OLIG 2 S147 A的smRNA导致更高和更稳定的蛋白质表达。使用单突变体0 LIG 2 smRNA形态原，我们建立了6天smRNA转染方案，并且神经胶质诱导导致从hiPSC快速产生NG 2 + OL祖细胞（OPC）（>70%纯度）。smRNA诱导的NG 2 + OPCs在体外可成熟为功能性OLs，在体内可促进髓鞘再生。总之，我们提出了一种安全有效的smRNA驱动的hiPSC分化为OL的策略，可用于神经退行性疾病患者的治疗性OPC/OL移植。合成修饰的信使RNA（smRNA）的使用允许人诱导的多能干细胞（hiPSC）分化成谱系特异性少突胶质细胞。
Transcription factors (TFs) have been introduced to drive the highly efficient differentiation of human-induced pluripotent stem cells (hiPSCs) into lineage-specific oligodendrocytes (OLs). However, effective strategies currently rely mainly on genome-integrating viruses. Here we show that a synthetic modified messenger RNA (smRNA)-based reprogramming method that leads to the generation of transgene-free OLs has been developed. An smRNA encoding a modified form of OLIG2, in which the serine 147 phosphorylation site is replaced with alanine, OLIG2S147A, is designed to reprogram hiPSCs into OLs. We demonstrate that repeated administration of the smRNA encoding OLIG2 S147A lead to higher and more stable protein expression. Using the single-mutant OLIG2 smRNA morphogen, we establish a 6-day smRNA transfection protocol, and glial induction lead to rapid NG2+ OL progenitor cell (OPC) generation (>70% purity) from hiPSC. The smRNA-induced NG2+ OPCs can mature into functional OLs in vitro and promote remyelination in vivo. Taken together, we present a safe and efficient smRNA-driven strategy for hiPSC differentiation into OLs, which may be utilized for therapeutic OPC/OL transplantation in patients with neurodegenerative disease. The use of synthetic modified messenger RNA (smRNA) allows for the differentiation of human-induced pluripotent stem cells (hiPSCs) into lineage-specific oligodendrocytes.
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影响因子：
5.3
作者：
Cayre M;Falque M;Mercier O;Magalon K;Durbec P
通讯作者：
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Kuhlmann T
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Verfaillie, Catherine M.
影响因子：
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通讯作者：
Farooq, M
影响因子：
4.8
作者：
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通讯作者：
Simons, Mikael