PRRX2 and HEY2 double knock-down facilitates ASCL1-induced neuron conversion in human dermal fibroblasts II

Forced expression of ASCL1, Nurr1, Lmx1a, miRNA-124 and p53shRNA (ANLmp) in fibroblasts reprograms fibroblasts to induced dopaminergic neurons (iDA). While human lung fibroblasts can be converted rapidly and efficiently, iDA of dermal fibroblast is very unefficient and incompleted. To address this issue,  we performed time series RNAseq on both lung and dermal fibroblasts during the first several days of ANLmp induced neuron convertion. Bioinformatics analysis revealed the stable fibroblast gene regulatory network (GRN) was a potential repressive factor for iDA in human dermal fibroblasts. Overall design: Time serial comparison of transcriptomes of non-isogenic human skin fibroblasts C002 and human lung fibroblasts MRC5 during their transdifferentiation to dopaminergic neuron induced by ANLmp.

在成纤维细胞（fibroblasts）中过表达ASCL1、Nurr1、Lmx1a、miRNA-124及p53shRNA（合称ANLmp），可将成纤维细胞重编程为诱导性多巴胺能神经元（induced dopaminergic neurons，iDA）。尽管人肺成纤维细胞可被快速且高效地重编程为iDA，但真皮成纤维细胞向iDA的转化效率极低且过程极不完整。为解决这一问题，我们在ANLmp诱导神经元转分化的最初数天内，对肺成纤维细胞和真皮成纤维细胞开展了时间序列RNA-seq（RNA测序）分析。生物信息学分析显示，稳定的成纤维细胞基因调控网络（gene regulatory network，GRN）是人真皮成纤维细胞向iDA转化的潜在抑制因素。实验整体设计：在ANLmp诱导非同基因人皮肤成纤维细胞C002与肺成纤维细胞MRC5向多巴胺能神经元转分化的过程中，对二者的转录组进行时间序列比对分析。