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

• Published in: Molecular biotechnology 2017-06, Vol.59 (6), p.207-220
• Main Authors: Ghazizadeh, Zaniar, Rassouli, Hassan, Fonoudi, Hananeh, Alikhani, Mehdi, Talkhabi, Mahmood, Darbandi-Azar, Amir, Chen, Shuibing, Baharvand, Hossein, Aghdami, Nasser, Salekdeh, Ghasem Hosseini
• Format: Article
• Language: English
• Subjects: 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
• ISSN: 1559-0305
• DOI: 10.1007/s12033-017-0007-x

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.