# Engineering multi-lineage inner ear organoids

> **NIH NIH R01** · BOSTON CHILDREN'S HOSPITAL · 2024 · $695,343

## Abstract

ABSTRACT
This research proposal focuses on the complex development of inner ear cells from various embryonic
lineages and the challenges of establishing multi-lineage inner ear tissues in vitro. The inability to routinely
utilize patient-derived inner ear explants due to their delicate nature prompts a necessity for a unique
approach. Thus, the project's long-term objective is to uncover the chemical and physical signals required to
cultivate functional inner ear tissue from human pluripotent stem cells (hPSCs) in vitro. Building on past
technological advancements, the research will refine a three-stage 3D culture system to create inner ear
organoids (IEOs) – a model that successfully generates sensory hair cells, neurons, glia, and mesenchymal
cells. The project seeks to overcome current limitations of IEO heterogeneity and free-floating nature, which
restrict live-cell imaging applications essential for advancing drug discovery and gene therapy testing. The
primary goal is to develop a standardized, reproducible IEO-on-a-chip system that mirrors the human fetal
inner ear, comprising sensory epithelia, peri-otic mesenchyme, and neural inputs. The project has three
specific goals: 1) refine the production and purification of otic organoid cellular components, 2) decipher the
physio-chemical needs for on-chip otic morphogenesis, and 3) establish image-based assays to investigate
hair cell function and dysfunction in IEOs-on-chip. We intend to use genomic data from every stage of IEO
development and leverage cell-cell communication analysis to enhance otic cell production. Next, we aim to
engineer an on-chip microenvironment promoting self-assembly of tubular otic sensory epithelia. Finally, the
study plans to establish image-based assays for on-chip assessment of genetic perturbation to hair cell
functional maturation using models related to the deafness-blindness disorder Usher Type 1. The successful
completion of this project would lead to a first-of-its-kind tissue chip for inner ear research, offering significant
value in the development of innovative gene therapies, as well as protective and regenerative drugs. The
proposed system shows potential in addressing current research challenges and spurring advancements in
understanding and treating hearing and balance-related diseases.

## Key facts

- **NIH application ID:** 10981671
- **Project number:** 2R01DC017461-07
- **Recipient organization:** BOSTON CHILDREN'S HOSPITAL
- **Principal Investigator:** Karl Russell Koehler
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2024
- **Award amount:** $695,343
- **Award type:** 2
- **Project period:** 2018-12-01 → 2029-06-30

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10981671

## Citation

> US National Institutes of Health, RePORTER application 10981671, Engineering multi-lineage inner ear organoids (2R01DC017461-07). Retrieved via AI Analytics 2026-05-24 from https://api.ai-analytics.org/grant/nih/10981671. Licensed CC0.

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