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Description

Hair cells (HCs) are the sensory cells of the inner ear responsible for detecting sound and movement information and sending it to the brain. Embryonic HC development relies on the transcriptional regulators ATOH1, POU4F3, and GFI1. Recently, analysis of GFI1 mutant HCs from both the cochlear (hearing) and vestibular (balance) systems revealed that GFI1 likely functions both as a transcriptional repressor to downregulate neuronal gene expression in early HCs, as well as a transcriptional activator in concert with ATOH1 to upregulate HC-specific genes. However, single cell-level transcriptional profiling of GFI1 mutant HCs has yet to be performed. This is likely due to the scarcity of HCs in the in vivo-derived inner ear for single cell RNA sequencing (scRNA-seq) analysis, as well as the sensitivity of mutant HCs to dissociation. Here, we use an inner ear organoid model to further study the role of GFI1 in HC development. Inner ear organoids produce sensory epithelia-like structures with thousands of vestibular-like HCs that can be used for developmental studies. Using scRNA-seq of later-stage inner ear organoids, we show that Gfi1 is expressed at the earliest stages of organoid HC formation (day in vitro (DIV) 11). We further validate the expression of GFI1 protein in organoid-derived HCs at DIV11, DIV16, and DIV21. In the future, we plan to utilize scRNA-seq of GFI1 CRISPRi mutant HCs to gain a better understanding of the specific genes GFI1 regulates during development. This research will be critical for establishing future regenerative therapies for inner ear dysfunction.

Publication Date

4-30-2026

Keywords

GFI1, organoids, hair cells

Investigating the Expression of the GFI1 Transcription Factor in Inner Ear Organoids

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