Transcription factor induced conversion of human fibroblasts towards the hair cell lineage

• Published in: PloS one Vol. 13; no. 7; p. e0200210
• Main Authors: Duran Alonso, María Beatriz, Lopez Hernandez, Iris, de la Fuente, Miguel Angel, Garcia-Sancho, Javier, Giraldez, Fernando, Schimmang, Thomas
• Format: Journal Article
• Language: English
• Published: United States Public Library of Science 06.07.2018; Public Library of Science (PLoS)
• Subjects: Acoustics; Basic Helix-Loop-Helix Transcription Factors - genetics; Basic Helix-Loop-Helix Transcription Factors - metabolism; Biology and Life Sciences; Biomarkers - metabolism; Brain; Cardiomyocytes; Cell lineage; Cell Lineage - drug effects; Cell Lineage - genetics; Cell Lineage - physiology; Cell Transdifferentiation - drug effects; Cell Transdifferentiation - genetics; Cell Transdifferentiation - physiology; Cells (biology); Cells, Cultured; Cochlea; Cèl·lules acústiques; Direct conversion; DNA-Binding Proteins - genetics; DNA-Binding Proteins - metabolism; Embryo cells; Embryonic stem cells; Epidermal Growth Factor - pharmacology; Epigenetics; Fibroblasts; Fibroblasts - cytology; Fibroblasts - drug effects; Fibroblasts - metabolism; Gene expression; Gene Expression - drug effects; Genes; Hair; Hair cells; Hair Cells, Auditory - cytology; Hair Cells, Auditory - drug effects; Hair Cells, Auditory - metabolism; Hearing loss; Homeodomain Proteins - genetics; Homeodomain Proteins - metabolism; Humans; Inhibitory postsynaptic potentials; Mammals; Math1 protein; Mechanoreceptors; Medicine and Health Sciences; Myosin VIIa; Myosins - genetics; Nanoparticles; Neonates; Pluripotency; Progenitor cells; Receptors; Recombinant Proteins - genetics; Recombinant Proteins - metabolism; Regeneration; Research and Analysis Methods; Restoration; Risk factors; RNA, Messenger - genetics; RNA, Messenger - metabolism; Somatic cells; Stem cells; Transcription activation; Transcription Factor Brn-3C - genetics; Transcription Factor Brn-3C - metabolism; Transcription factors; Transcription Factors - genetics; Transcription Factors - metabolism; Tretinoin - pharmacology; United States > US; Grand Island New York; Spain
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0200210

Hearing loss is the most common sensorineural disorder, affecting over 5% of the population worldwide. Its most frequent cause is the loss of hair cells (HCs), the mechanosensory receptors of the cochlea. HCs transduce incoming sounds into electrical signals that activate auditory neurons, which in turn send this information to the brain. Although some spontaneous HC regeneration has been observed in neonatal mammals, the very small pool of putative progenitor cells that have been identified in the adult mammalian cochlea is not able to replace the damaged HCs, making any hearing impairment permanent. To date, guided differentiation of human cells to HC-like cells has only been achieved using either embryonic stem cells (ESCs) or induced pluripotent stem cells (iPSCs). However, use of such cell types suffers from a number of important disadvantages, such as the risk of tumourigenicity if transplanted into the host´s tissue. We have obtained cells expressing hair cell markers from cultures of human fibroblasts by overexpression of GFI1, Pou4f3 and ATOH1 (GPA), three genes that are known to play a critical role in the development of HCs. Immunocytochemical, qPCR and RNAseq analyses demonstrate the expression of genes typically expressed by HCs in the transdifferentiated cells. Our protocol represents a much faster approach than the methods applied to ESCs and iPSCs and validates the combination of GPA as a set of genes whose activation leads to the direct conversion of human somatic cells towards the hair cell lineage. Our observations are expected to contribute to the development of future therapies aimed at the regeneration of the auditory organ and the restoration of hearing.