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Generation of Inner Ear Hair Cells by Direct Lineage Conversion of Primary Somatic Cells

ABSTRACT: This SuperSeries is composed of the SubSeries listed below.

ORGANISM(S): Mus musculus

PROVIDER: GSE149260 | GEO | 2020/05/28

REPOSITORIES: GEO

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Generation of inner ear hair cells by direct lineage conversion of primary somatic cells.

Project description:The mechanoreceptive sensory hair cells in the inner ear are selectively vulnerable to numerous genetic and environmental insults. In mammals, hair cells lack regenerative capacity, and their death leads to permanent hearing loss and vestibular dysfunction. Their paucity and inaccessibility has limited the search for otoprotective and regenerative strategies. Growing hair cells in vitro would provide a route to overcome this experimental bottleneck. We report a combination of four transcription factors (Six1, Atoh1, Pou4f3, and Gfi1) that can convert mouse embryonic fibroblasts, adult tail-tip fibroblasts and postnatal supporting cells into induced hair cell-like cells (iHCs). iHCs exhibit hair cell-like morphology, transcriptomic and epigenetic profiles, electrophysiological properties, mechanosensory channel expression, and vulnerability to ototoxin in a high-content phenotypic screening system. Thus, direct reprogramming provides a platform to identify causes and treatments for hair cell loss, and may help identify future gene therapy approaches for restoring hearing.

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Generation of Inner Ear Hair Cells by Direct Lineage Conversion of Primary Somatic Cells [ChIP-seq]

Project description:We purified seven different cell populations and performed RNA sequencing to profile transcriptional similarities and differences between them. The seven cell types were 1) Atoh1-GFP positive cochlear hair cells from the organ of Corti of postnatal day one mice, 2) Atoh1-GFP negative cells from the organ of Corti of postnatal day one mice, 3) Atoh1-GFP positive induced hair cells generated by overexpression of Six1, Atoh1, Pou4f3, and Gfi1, 4) dsRed transduced control mouse embryonic fibroblasts, 5) Atoh1-GFP positive Merkel cells from postnatal day 1 mice, 6) Atoh1-GFP positive Cerebellar granule precursor cells from postnatal day 1 mice, and 7) Atoh1-GFP positive Gut secretory cells from postnatal day one mice.

2020-05-28 | GSE149259 | GEO

Generation of Inner Ear Hair Cells by Direct Lineage Conversion of Primary Somatic Cells [ATAC-seq]

2020-05-28 | GSE149258 | GEO

Generation of Inner Ear Hair Cells by Direct Lineage Conversion of Primary Somatic Cells [RNA-seq]

2020-05-28 | GSE149257 | GEO

Generation of Inner Ear Hair Cells by Direct Lineage Conversion of Primary Somatic Cells

Project description:Generation of Inner Ear Hair Cells by Direct Lineage Conversion of Primary Somatic Cells

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Generation of Inner Ear Hair Cells by Direct Lineage Conversion of Primary Somatic Cells [ATAC-seq]

Project description:Generation of Inner Ear Hair Cells by Direct Lineage Conversion of Primary Somatic Cells [ATAC-seq]

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Generation of Inner Ear Hair Cells by Direct Lineage Conversion of Primary Somatic Cells [ChIP-seq]

Project description:Generation of Inner Ear Hair Cells by Direct Lineage Conversion of Primary Somatic Cells [ChIP-seq]

| PRJNA627750 | ENA

Generation of Inner Ear Hair Cells by Direct Lineage Conversion of Primary Somatic Cells [RNA-seq]

Project description:Generation of Inner Ear Hair Cells by Direct Lineage Conversion of Primary Somatic Cells [RNA-seq]

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Generation of oligodendroglial cells by direct lineage conversion.

Project description:Transplantation of oligodendrocyte precursor cells (OPCs) is a promising potential therapeutic strategy for diseases affecting myelin. However, the derivation of engraftable OPCs from human pluripotent stem cells has proven difficult and primary OPCs are not readily available. Here we report the generation of induced OPCs (iOPCs) by direct lineage conversion. Forced expression of the three transcription factors Sox10, Olig2 and Zfp536 was sufficient to reprogram mouse and rat fibroblasts into iOPCs with morphologies and gene expression signatures resembling primary OPCs. More importantly, iOPCs gave rise to mature oligodendrocytes that could ensheath multiple host axons when co-cultured with primary dorsal root ganglion cells and formed myelin after transplantation into shiverer mice. We propose direct lineage reprogramming as a viable alternative approach for the generation of OPCs for use in disease modeling and regenerative medicine.

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Generation of inner ear organoids containing functional hair cells from human pluripotent stem cells.

Project description:The derivation of human inner ear tissue from pluripotent stem cells would enable in vitro screening of drug candidates for the treatment of hearing and balance dysfunction and may provide a source of cells for cell-based therapies of the inner ear. Here we report a method for differentiating human pluripotent stem cells to inner ear organoids that harbor functional hair cells. Using a three-dimensional culture system, we modulate TGF, BMP, FGF, and WNT signaling to generate multiple otic-vesicle-like structures from a single stem-cell aggregate. Over 2 months, the vesicles develop into inner ear organoids with sensory epithelia that are innervated by sensory neurons. Additionally, using CRISPR-Cas9, we generate an ATOH1-2A-eGFP cell line to detect hair cell induction and demonstrate that derived hair cells exhibit electrophysiological properties similar to those of native sensory hair cells. Our culture system should facilitate the study of human inner ear development and research on therapies for diseases of the inner ear.

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