Single Cell QPCR Reveals That Additional HAND2 and MicroRNA-1 Facilitate the Early Reprogramming Progress of Seven-factor-induced Human Myocytes

Overview

Journal PLoS One

Specialties General Medicine
Science

Date 2017 Aug 11

PMID 28796841

Citations 11

Authors

Emre Bektik

Adrienne Dennis

Prateek Prasanna

Anant Madabhushi

Ji-Dong Fu

Affiliations

Soon will be listed here.

Abstract

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.

Citing Articles

MicroRNA-1 Deficiency Is a Primary Etiological Factor Disrupting Cardiac Contractility and Electrophysiological Homeostasis.

Yang D, Wan X, Schwieterman N, Cavus O, Kacira E, Xu X Circ Arrhythm Electrophysiol. 2023; 17(1):e012150.

PMID: 38126205 PMC: 10842700. DOI: 10.1161/CIRCEP.123.012150.

Direct Cardiac Reprogramming: Current Status and Future Prospects.

Haridhasapavalan K, Borthakur A, Thummer R Adv Exp Med Biol. 2023; 1436:1-18.

PMID: 36662416 DOI: 10.1007/5584_2022_760.

Improving Cardiac Reprogramming for Heart Regeneration in Translational Medicine.

Liu L, Guo Y, Li Z, Wang Z Cells. 2021; 10(12).

PMID: 34943805 PMC: 8699771. DOI: 10.3390/cells10123297.

Fibroblast transition to an endothelial "trans" state improves cell reprogramming efficiency.

Mathison M, Sanagasetti D, Singh V, Pugazenthi A, Pinnamaneni J, Ryan C Sci Rep. 2021; 11(1):22605.

PMID: 34799643 PMC: 8604927. DOI: 10.1038/s41598-021-02056-x.

Inhibition of CREB-CBP Signaling Improves Fibroblast Plasticity for Direct Cardiac Reprogramming.

Bektik E, Sun Y, Dennis A, Sakon P, Yang D, Deschenes I Cells. 2021; 10(7).

PMID: 34206684 PMC: 8307124. DOI: 10.3390/cells10071572.

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