# A Human Enteroid Model of Cholera Toxin Pathophysiology

> **NIH NIH K01** · JOHNS HOPKINS UNIVERSITY · 2020 · $158,220

## Abstract

PROJECT SUMMARY/ABSTRACT
Vibrio cholerae infection continues to be a life-threatening concern in regions with crowded living conditions
and poor sanitation. The emergence of El Tor variant, a strain that exhibits enhanced cholera toxin (CT)
production and innate immune activation, necessitates examination of the factors that contribute to increased
disease severity in recent outbreaks. CT elicits production and secretion of prostaglandin E2 (PGE2), serotonin
(5-HT), and numerous cytokines that may not only regulate intestinal fluid transport, but may also contribute to
the innate immune response. In our novel observation, the CT B subunit was found to induce cell cycle arrest
in actively dividing intestinal stem/progenitor cells, and we hypothesize that this phenomenon may arise from
the same signaling that yields PGE2, 5-HT, cAMP, or cytokine release. To facilitate mechanistic studies of CT-
induced biomolecule secretion and cell cycle arrest, we will employ primary untransformed human enteroids
derived from adult stem cells of the intestine. We have developed a method to grow human enteroids as 2-
dimensional epithelial monolayers, overcoming the limitation of 3-dimensional cultures that prevents direct
access to the polarized apical cell surface. This K01 proposal will establish understanding of CT-mediated
innate immune response and cell cycle dysregulation using the enteroid monolayer model in the following
aims: We will 1) identify how CT induces PGE2 synthesis and IL-8 secretion through CRISPR/Cas9-
guided receptor knock-outs, pharmacological inhibitor studies, and secreted PGE2 and cytokine ELISA. Using
a recently reported method to enrich enteroid cultures with the enterochromaffin cell lineage, we will 2)
determine how CT increases extracellular 5-HT and how this contributes to cytokine release. This will
include evaluation of direct or indirect stimulation of 5-HT secretion, function of the serotonin transporter
(SERT), and enterocyte 5-HT receptor roles in cytokine production. Finally, we will 3) determine the origin
and duration of CT-induced cell cycle arrest using EdU incorporation, FACS analysis of cell cycle phase,
and RNA-Seq to compare transcriptional changes induced by CT, CTB, PGE2, 5-HT, cAMP, and specific
cytokines. Not only will these aims advance understanding of CT-induced pathophysiology with potential
clinical significance, but they will also foster technical skill development in cytokine detection/quantitation,
CRISPR/Cas9-driven gene editing, and computational bioinformatics to analyze RNA-Seq data. My advisory
committee of highly qualified scientific and professional role models and the resource-rich environment in the
Division of Gastroenterology at the Johns Hopkins University School of Medicine will guide my development
from a mentored researcher to an independent scientist in the field of diarrheal diseases and intestinal stem
cell effects caused by enteric microbial pathogens.

## Key facts

- **NIH application ID:** 9830641
- **Project number:** 5K01DK113043-03
- **Recipient organization:** JOHNS HOPKINS UNIVERSITY
- **Principal Investigator:** Jennifer Foulke-Abel
- **Activity code:** K01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2020
- **Award amount:** $158,220
- **Award type:** 5
- **Project period:** 2018-01-09 → 2022-11-30

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/9830641

## Citation

> US National Institutes of Health, RePORTER application 9830641, A Human Enteroid Model of Cholera Toxin Pathophysiology (5K01DK113043-03). Retrieved via AI Analytics 2026-05-23 from https://api.ai-analytics.org/grant/nih/9830641. Licensed CC0.

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