# Autologous cell transplantation for the treatment of colorectal aganglionosis

> **NIH NIH R03** · MASSACHUSETTS GENERAL HOSPITAL · 2021 · $77,472

## Abstract

PROJECT SUMMARY
Hirschsprung disease is a potentially lethal congenital disorder characterized by the absence of
enteric nervous system (ENS) along variable lengths of distal intestine due to the failure of
neural crest-derived cells to colonize the entire intestine during development. The surgical
treatment of Hirschsprung disease involves removing the aganglionic segment. While this is life-
saving, significant complications commonly occur after surgery, including constipation, fecal
incontinence, and enterocolitis. Transplantation of enteric neural crest-derived cells (ENCCs)
offers a promising new approach to replacing missing or abnormal enteric neurons in
Hirschsprung disease and other neurointestinal diseases. Recent studies, including from our
laboratory, demonstrate that ENCCs can be isolated from the postnatal intestine, propagated in
culture, and transplanted into the gut wall. However, major challenges remain: (1) the numbers
of neurons generated have been limited, (2) their ability to integrate into neuroglial networks
capable of restoring gut function has yet to be demonstrated, and (3) mice with Hirschsprung
disease die at 4-6 weeks of age, limiting the time window available for analysis after cell
transplantation. To overcome these challenges, we propose the following aims: (1) to
incorporate an innovative approach to isolating and expanding postnatal gut-derived ENCCs
that generates greater numbers of progenitor ENCCs than prior methods and (2) to transplant
autologously-derived ENCCs into a novel non-lethal model of colorectal aganglionosis
generated by local injection of human diphtheria toxin (DT) into transgenic mice whose neural
crest-derived cells express DT receptor. To optimize ENCC expansion, a combinatorial drug
screen approach will be used. ENCCs will be cultured with candidate molecules, together with
known growth factors, to identify the optimal “cocktail” for progenitor cell expansion. Single cell
RNAseq will be performed to characterize the transcriptome profile of cells prior to
transplantation. Autologously-derived donor cells will be delivered into the experimentally-
generated aganglionic segment of colon. Analyses will include quantitative determination of
ENCC survival, proliferation, and neuronal differentiation, and functional characterization of
neuronal activity. The results obtained will establish an optimized approach to ENCC isolation,
expansion, and transplantation, and demonstrate the potential of cell-based therapy to restore
GI function in Hirschsprung disease and other neurointestinal diseases.

## Key facts

- **NIH application ID:** 10128804
- **Project number:** 1R03HD100762-01A1
- **Recipient organization:** MASSACHUSETTS GENERAL HOSPITAL
- **Principal Investigator:** Ryo Hotta
- **Activity code:** R03 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2021
- **Award amount:** $77,472
- **Award type:** 1
- **Project period:** 2020-12-01 → 2022-11-30

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10128804

## Citation

> US National Institutes of Health, RePORTER application 10128804, Autologous cell transplantation for the treatment of colorectal aganglionosis (1R03HD100762-01A1). Retrieved via AI Analytics 2026-05-23 from https://api.ai-analytics.org/grant/nih/10128804. Licensed CC0.

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