# Investigating Novel Methods to Combat Urinary Tract Infections

> **NIH NIH K99** · UNIVERSITY OF WASHINGTON · 2022 · $99,999

## Abstract

PROJECT SUMMARY/ABSTRACT
Urinary Tract Infections (UTIs) are common in children, the elderly, diabetics, and immunocompromised
patients. UTIs have become difficult to treat due to the rise in multi-drug resistant bacteria such as the
extended spectrum beta lactamase (ESBL) producing Escherichia coli and the carbapenem resistant Klebsiella
pneumoniae. Therefore, there is an urgent need to develop alternative treatments, for instance, anti-adhesive
molecules that block bacteria adhesion to host cells, the all important first step in initiating an infection.
Pathogenic bacteria, like E. coli and K. pneumoniae establish infections in the urinary tract by adhering to host
epithelial cells via adhesion proteins, such as FimH, FmlH and MrkD. The long-term objective of this
proposal is to develop novel methods of inhibiting bacterial adhesion. A monoclonal antibody (mAb) that
inhibits FimH-mediated bacterial adhesion has been discovered, but its mechanism of action has not been
investigated. Moreover, previous studies show that this mAb is neither a competitive nor an allosteric inhibitor,
indicating a novel mechanism of biomolecular inhibition. The aim of this proposal is to elucidate the mechanism
by which the mAb inhibits FimH-mediated bacterial adhesion and develop molecules capable of inhibiting FimH
function. Aim 1 studies will investigate the mechanism of mAb inhibition by mapping the binding site of
the mAb on FimH using Nuclear Magnetic Resonance Spectroscopy (NMR) and Hydrogen Deuterium
Exchange Mass Spectrometry (HDX-MS) and determining the X-ray crystal structure of the FimH-mAb
complex. Furthermore, NMR and HDX-MS will provide information on how mAb binding alters the
conformational dynamics of FimH. These structural and dynamics studies will provide insights into the mAb
mechanism of action. In Aim 2, peptides of the mAb that interact with FimH will be identified using HDX-
MS and tested for binding and inhibitory function in bacterial adhesion assays. The binding and inhibitory
function of the mAb peptides will be optimized using Deep Mutational Scanning (DMS) and Rosetta
computational peptide design. In Aim 3, the structures, and dynamics of FmlH and MrkD will be
investigated, and peptide inhibitors will be developed. Mutagenesis and NMR studies will be carried out to
investigate the different structural conformations and dynamics of FmlH and MrkD. Finally, computational
peptide design using Rosetta will be employed in the de novo design of peptide inhibitors against FmlH and
MrkD.

## Key facts

- **NIH application ID:** 10369290
- **Project number:** 1K99GM141364-01A1
- **Recipient organization:** UNIVERSITY OF WASHINGTON
- **Principal Investigator:** PEARL MAGALA
- **Activity code:** K99 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2022
- **Award amount:** $99,999
- **Award type:** 1
- **Project period:** 2022-09-16 → 2024-09-15

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10369290

## Citation

> US National Institutes of Health, RePORTER application 10369290, Investigating Novel Methods to Combat Urinary Tract Infections (1K99GM141364-01A1). Retrieved via AI Analytics 2026-05-26 from https://api.ai-analytics.org/grant/nih/10369290. Licensed CC0.

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