PTBP2 attenuation facilitates fibroblast to neuron conversion by promoting alternative splicing of neuronal genes.
PTBP2 attenuation facilitates fibroblast to neuron conversion by promoting alternative splicing of neuronal genes.
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DOI：
10.1016/j.stemcr.2023.09.012
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发表时间：
2023-11-14
影响因子：
5.9
通讯作者：
Feng, Jian
中科院分区：
文献类型：
--
作者：
Zhu, Binglin;Fisher, Emily;Li, Li;Zhong, Ping;Yan, Zhen;Feng, Jian
人皮肤成纤维细胞直接转化为神经元的效率较低，机制尚不清楚。在这里，我们发现，PTBP2基因的敲除显著增强了ASCL1、MIR9/9、∗-124和P53shRNA(Amp)诱导的转分化，产生了主要的GABA能神经元。纵向RNA测序分析证实了许多RNA剪接调控因子的持续诱导。其中，RBFOX3(Neun)基因的敲除显著抑制了细胞的转分化。过表达RBFOX3可显著增强Amp诱导的转化率，这种增强作用可被PTBP2基因敲除所阻断。我们发现，PTBP2的衰减显著有利于参与突触传递、信号转导和轴突形成的许多基因的神经元特异性选择性剪接(AS)。RBFOX3基因敲除显著逆转了这种作用，而RBFOX3过表达则阻断了这种增强。这项研究揭示了神经元特异性AS在人皮肤成纤维细胞直接转化为神经元过程中的关键作用，表明PTBP2衰减与RBFOX3协同增强了这一机制。PTBP2基因敲除增强了ASCL1介导的人成纤维细胞向神经元的转化，成纤维细胞向神经元的转化诱导RSR，包括RBFOX3基因敲除或过表达，PTBP2基因敲除或阻断PTBP2基因敲除的作用促进神经元特异性，与RBFOX3协同，人皮肤成纤维细胞向神经元的直接转化效率低，机制尚不清楚。冯和他的同事们发现，PTBP2(NPTB)基因敲除显著增强了ASCL1、MIR9/9、∗-124和P53shRNA(Amp)诱导的转分化，主要产生GABA能神经元。AMNP诱导RBFOX3，RBFOX3促进神经元特异性选择性剪接，与nPTB基因敲除协同促进转分化。
The direct conversion of human skin fibroblasts to neurons has a low efficiency and unclear mechanism. Here, we show that the knockdown of PTBP2 significantly enhanced the transdifferentiation induced by ASCL1, MIR9/9∗-124, and p53 shRNA (AMp) to generate mostly GABAergic neurons. Longitudinal RNA sequencing analyses identified the continuous induction of many RNA splicing regulators. Among these, the knockdown of RBFOX3 (NeuN), significantly abrogated the transdifferentiation. Overexpression of RBFOX3 significantly enhanced the conversion induced by AMp; the enhancement was occluded by PTBP2 knockdown. We found that PTBP2 attenuation significantly favored neuron-specific alternative splicing (AS) of many genes involved in synaptic transmission, signal transduction, and axon formation. RBFOX3 knockdown significantly reversed the effect, while RBFOX3 overexpression occluded the enhancement. The study reveals the critical role of neuron-specific AS in the direct conversion of human skin fibroblasts to neurons by showing that PTBP2 attenuation enhances this mechanism in concert with RBFOX3. PTBP2 knockdown enhances ASCL1-mediated conversion of human fibroblasts to neurons The fibroblast-to-neuron conversion induces RSRs including RBFOX3 RBFOX3 knockdown or overexpression ablates or occludes the effect of PTBP2 knockdown PTBP2 knockdown promotes neuron-specific AS in concert with RBFOX3 The direct conversion of human skin fibroblasts to neurons has a low efficiency and unclear mechanism. Here, Feng and colleagues show that PTBP2 (nPTB) knockdown significantly enhanced the transdifferentiation induced by ASCL1, MIR9/9∗-124, and p53 shRNA (AMp) to generate mostly GABAergic neurons. AMnp induced RBFOX3, which promoted neuron-specific alternative splicing in concert with nPTB knockdown to enhance the transdifferentiation.
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