# Engineering multi-lineage human inner ear organoids

> **NIH NIH R01** · BOSTON CHILDREN'S HOSPITAL · 2021 · $528,331

## Abstract

ABSTRACT
Inner ear development requires the assembly of diverse cells from multiple embryonic lineages. The epithelial,
neuronal, and glial components of the inner ear are ectoderm-derived, whereas the mesenchymal components
are predominantly mesoderm-derived. A major engineering challenge is to establish multi-lineage inner ear
tissues in vitro, which researchers could use to study human hearing and balance-related diseases, investigate
developmental biology questions, and evaluate promising therapeutics. The routine use of patient-derived
inner ear explants for research is not feasible because the human inner ear is difficult to biopsy. Therefore, our
long-term goal is to define the chemical and physical signals required to recapitulate formation of functional
human inner ear tissue in vitro from human pluripotent stem cells (hPSCs). This project builds upon a recent
technological innovation reported by our laboratory: a multi-stage 3D culture system for generating inner ear
organoids that contain sensory hair cells and neurons. Despite significant progress, there are remaining
questions about how faithfully inner ear organoids mimic normal embryonic development. Moreover, there are
technical hurdles that may limit integration of inner ear organoids into tissue-chip drug discovery platforms.
Specifically, organoid production efficiency is variable and the full range of cell types in organoids is unclear.
Moreover, our ability to track the development or physiology of inner ear sensory cells in real-time is limited.
Our research plan will define a next-generation inner ear organoid system. For Aim 1, we will use high-
throughput single-cell analysis to generate a cell fate map of developing inner ear organoids. In Aim 2, we will
generate dual-reporter hPSC lines for real-time monitoring of inner ear organoid sensorineural networks. In
Aim 3, we will engineer chemically-defined inner ear organoids with improved fidelity to mammalian
development. Finally, we will verify inner ear organoid production from a set of four human induced pluripotent
stem cell lines to ensure the reproducibility of our results. Together, completion of this project will deepen our
characterization of the human inner ear organoid model and facilitate transfer of the technology to other
research laboratories. Future investigations could pursue unexplored cell signaling mechanisms, model genetic
diseases, or integrate organoids into tissue-chip systems. We anticipate that our study will provide broadly
applicable insights that should aid the production of organoids of other sensory systems and should provide a
powerful tool for otolaryngology research.

## Key facts

- **NIH application ID:** 10067370
- **Project number:** 5R01DC017461-04
- **Recipient organization:** BOSTON CHILDREN'S HOSPITAL
- **Principal Investigator:** Karl Russell Koehler
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2021
- **Award amount:** $528,331
- **Award type:** 5
- **Project period:** 2018-12-01 → 2023-11-30

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10067370

## Citation

> US National Institutes of Health, RePORTER application 10067370, Engineering multi-lineage human inner ear organoids (5R01DC017461-04). Retrieved via AI Analytics 2026-05-23 from https://api.ai-analytics.org/grant/nih/10067370. Licensed CC0.

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