Single cell qPCR reveals that additional HAND2 and microRNA-1 facilitate the early reprogramming progress of seven-factor-induced human myocytes.
Single cell qPCR reveals that additional HAND2 and microRNA-1 facilitate the early reprogramming progress of seven-factor-induced human myocytes.
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DOI：
10.1371/journal.pone.0183000
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发表时间：
2017
期刊：
影响因子：
3.7
通讯作者：
Fu JD
中科院分区：
文献类型：
--
作者：
Bektik E;Dennis A;Prasanna P;Madabhushi A;Fu JD
将心脏成纤维细胞直接重编程为诱导的心肌细胞（CM）样细胞（iCM）在恢复心脏功能方面具有很大的前景。我们以前发现，人成纤维细胞可以通过7个重编程因子向CM样细胞重编程;然而，人成纤维细胞中的iCM重编程比小鼠细胞中的iCM重编程更困难，也更耗时。在这项研究中，我们研究了额外的重编程因子是否可以通过单细胞定量PCR定量和/或定性地改善7因子介导的人iCM重编程。我们首先验证了46对TaqMan®引物/探针，这些引物/探针具有足够的效率和灵敏度来检测单个H9人胚胎干细胞（ESC）分化的CM（H9 CM）和人成纤维细胞之间基因表达的显着差异。这46个基因的表达谱显示，与通过7个因子重编程的4周iCM相比，12周iCM中的重编程有所改善，表明人iCM重编程期间的随机期延长。尽管没有一个额外的重编程因子产生更多数量的iCM，但我们的单细胞qPCR显示，额外的HAND 2或microRNA-1可以促进成纤维细胞基因的沉默，并在更多的重编程iCM中产生更好程度的重编程。值得注意的是，HAND 2表达越多，7 Fs + HAND 2重编程iCM中激活的心脏基因水平越高。总之，HAND 2和microRNA-1可以帮助7个因子促进人成纤维细胞iCM重编程的早期进展。我们的研究为进一步优化人类细胞中直接iCM重编程的方法提供了有价值的信息。
The direct reprogramming of cardiac fibroblasts into induced cardiomyocyte (CM)-like cells (iCMs) holds great promise in restoring heart function. We previously found that human fibroblasts could be reprogrammed toward CM-like cells by 7 reprogramming factors; however, iCM reprogramming in human fibroblasts is both more difficult and more time-intensive than that in mouse cells. In this study, we investigated if additional reprogramming factors could quantitatively and/or qualitatively improve 7-factor-mediated human iCM reprogramming by single-cell quantitative PCR. We first validated 46 pairs of TaqMan® primers/probes that had sufficient efficiency and sensitivity to detect the significant difference of gene expression between individual H9 human embryonic stem cell (ESC)-differentiated CMs (H9CMs) and human fibroblasts. The expression profile of these 46 genes revealed an improved reprogramming in 12-week iCMs compared to 4-week iCMs reprogrammed by 7 factors, indicating a prolonged stochastic phase during human iCM reprogramming. Although none of additional one reprogramming factor yielded a greater number of iCMs, our single-cell qPCR revealed that additional HAND2 or microRNA-1 could facilitate the silencing of fibroblast genes and yield a better degree of reprogramming in more reprogrammed iCMs. Noticeably, the more HAND2 expressed, the higher-level were cardiac genes activated in 7Fs+HAND2-reprogrammed iCMs. In conclusion, HAND2 and microRNA-1 could help 7 factors to facilitate the early progress of iCM-reprogramming from human fibroblasts. Our study provides valuable information to further optimize a method of direct iCM-reprogramming in human cells.
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影响因子：
64.8
作者：
Qian, Li;Huang, Yu;Srivastava, Deepak
通讯作者：
Srivastava, Deepak
影响因子：
5
作者：
Protze, Stephanie;Khattak, Shahryar;Ravens, Ursula
通讯作者：
Ravens, Ursula
通过对心脏转录因子的非肌细胞进行重编程通过重编程。
影响因子：
64.8
作者：
Song, Kunhua;Nam, Young-Jae;Luo, Xiang;Qi, Xiaoxia;Tan, Wei;Huang, Guo N.;Acharya, Asha;Smith, Christopher L.;Tallquist, Michelle D.;Neilson, Eric G.;Hill, Joseph A.;Bassel-Duby, Rhonda;Olson, Eric N.
通讯作者：
Olson, Eric N.
DOI：
10.1146/annurev-cellbio-101011-155739
发表时间：
2012
影响因子：
11.3
作者：
Kikuchi K;Poss KD
通讯作者：
Poss KD
影响因子：
5
作者：
Addis RC;Ifkovits JL;Pinto F;Kellam LD;Esteso P;Rentschler S;Christoforou N;Epstein JA;Gearhart JD
通讯作者：
Gearhart JD