Journal Article

Transient Activation of Reprogramming Transcription Factors Using Protein Transduction Facilitates Conversion of Human Fibroblasts Toward Cardiomyocyte-Like Cells

Fonoudi, Hananeh, Ghazizadeh, Zaniar, Rassouli, Hassan, Darbandi-Azar, Amir, Salekdeh, Ghasem Hosseini, Talkhabi, Mahmood, Baharvand, Hossein, Alikhani, Mehdi, Chen, Shuibing, Aghdami, Nasser

2017

Overview

Derivation of cardiomyocytes directly from patients' own fibroblasts could offer a new therapeutic approach for those with ischemic heart disease. An essential step toward clinical application is to establish safe conversion of human fibroblasts into a cardiac fate. Here we aimed to efficiently and safely generate cardiomyocytes from human fibroblasts by direct delivery of reprogramming recombinant cell permeant form of reprogramming proteins followed by cardio-inductive signals. Human fetal and adult fibroblasts were transiently exposed to transactivator of transcription-fused recombinant OCT4, SOX2, KLF4 and c-MYC for 2 weeks and then were directly differentiated toward protein-induced cardiomyocyte-like cells (p-iCLCs) in a cardiac fate niche, carried out by treatment with a set of cardiogenic small molecules (sequential treatment of Chir, and IWP-2, SB431542 and purmorphamine). The cells showed cardiac phenotype over a period of 3 weeks without first undergoing reprogramming into or through a pluripotent intermediate, shown by lack of expression of key pluripotency markers. p-iCLCs exhibited cardiac features at both the gene and protein levels. Our study provides an alternative method for the generation of p-iCLCs which shortcut reprogramming toward allogeneic cardiomyocytes in a safe and efficient manner and could facilitate generation of genetic material-free cardiomyocytes.

• Subject: Cellular Reprogramming - genetics / Cellular Reprogramming - physiology / Fibroblasts - metabolism / Humans / Kruppel-Like Factor 4 / Kruppel-Like Transcription Factors - genetics / Kruppel-Like Transcription Factors - metabolism / Myocytes, Cardiac - metabolism / Octamer Transcription Factor-3 - genetics / Octamer Transcription Factor-3 - metabolism / Proto-Oncogene Proteins c-myc - genetics / Proto-Oncogene Proteins c-myc - metabolism / Recombinant Proteins - genetics / Recombinant Proteins - metabolism / SOXB1 Transcription Factors - genetics / SOXB1 Transcription Factors - metabolism / Transcription Factors - genetics / Transcription Factors - metabolism