# Project 1: Pathogenesis of Enteroaggregative E. coli using human colonoids

> **NIH NIH P01** · UNIVERSITY OF MARYLAND BALTIMORE · 2020 · $268,600

## Abstract

Abstract
Enteroaggregative E. coli (EAEC) is an important pathogen of traveler's diarrhea, diarrhea in industrialized
countries and growth faltering in developing countries. In this application, we propose to study EAEC
pathogenesis in the human intestinal mucosa through the use of newly established human enteroid/colonoid
model, which is an ex vivo self–propagating human intestinal primary culture. This application comprises a
consortium among investigators at Johns Hopkins University (JHU), who have pioneered the development of
the enteroid/colonoid model, and researchers at the University of Maryland (UMD) and the University of
Virginia, who are leaders in the study of the pathogenesis of diarrheal disease cause by diarrhegenic E. coli
and Shigella. This project will leverage the existence of the new colonoid monolayer model. We will focus on
characterization of EAEC interactions with colonoid and colonoids/leukocyte models, in particular to dissect the
roles of AAFs, which have emerged as the principle EAEC virulence factor in multiple models, and the function
of SPATEs, which have shown broad spectrum activities on mucin and leukocyte glycoproteins, promoting
immunomodulatory effects. This project will comprise two Specific Aims. In Aim 1, we will characterize
interactions of EAEC with the intestinal mucosa. We will extend ongoing studies which suggest that EAEC
binding to MUC glycoproteins is fundamental to its pathogenesis. We will extend structure-function studies of
the AAF adhesin and illuminate the contribution of AAF basic residues to interaction of EAEC with the human
intestinal mucosa. We will determine the contribution of AAF mucin-binding residues to EAEC interaction with
the human mucosa. We will characterize the contributions of Pic and dispersin to AAF-mediated interaction
with the mucosa using time-lapse microscopy methods. In Aim 2 we will characterize the immunomodulatory
role of class 2 SPATEs on the intestinal mucosa. We will address the potential roles of cleavage of intestinal
glycoproteins in EAEC pathogenesis. We will dissect the inflammatory response through binding and cleavage
of O-linked glycoproteins such as the MUC signaling proteins on the intestinal mucosa. We will also
characterize the immunomodulatory effect of Pic through cleavage of inflammatory cytokine receptors in the
colonoid/macrophage system. The work described here will continue to advance the general study of EAEC
pathogenesis, generating fundamental insights that will illuminate aspects of pathogenesis relevant to other
enteric pathogens. We will work closely and synergistically with investigators in other projects and the Core
components.

## Key facts

- **NIH application ID:** 9982181
- **Project number:** 5P01AI125181-05
- **Recipient organization:** UNIVERSITY OF MARYLAND BALTIMORE
- **Principal Investigator:** James P. Nataro
- **Activity code:** P01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2020
- **Award amount:** $268,600
- **Award type:** 5
- **Project period:** — → 2021-06-30

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/9982181

## Citation

> US National Institutes of Health, RePORTER application 9982181, Project 1: Pathogenesis of Enteroaggregative E. coli using human colonoids (5P01AI125181-05). Retrieved via AI Analytics 2026-05-23 from https://api.ai-analytics.org/grant/nih/9982181. Licensed CC0.

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