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A Semi-Quantitative Assay to Measure Glycosaminoglycan Degradation by the Urinary Microbiota

Glycosaminoglycans (GAGs) are linear polysaccharides and are among the primary components of mucosal surfaces in mammalian systems. The GAG layer lining the mucosal surface of the urinary tract is thought to play a critical role in urinary tract homeostasis and provide a barrier against urinary trac...

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Autores principales: Nguyen, Vivian H., Khan, Fatima, Shipman, Braden M., Neugent, Michael L., Hulyalkar, Neha V., Cha, Natalie Y., Zimmern, Philippe E., De Nisco, Nicole J.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8762050/
https://www.ncbi.nlm.nih.gov/pubmed/35047421
http://dx.doi.org/10.3389/fcimb.2021.803409
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author Nguyen, Vivian H.
Khan, Fatima
Shipman, Braden M.
Neugent, Michael L.
Hulyalkar, Neha V.
Cha, Natalie Y.
Zimmern, Philippe E.
De Nisco, Nicole J.
author_facet Nguyen, Vivian H.
Khan, Fatima
Shipman, Braden M.
Neugent, Michael L.
Hulyalkar, Neha V.
Cha, Natalie Y.
Zimmern, Philippe E.
De Nisco, Nicole J.
author_sort Nguyen, Vivian H.
collection PubMed
description Glycosaminoglycans (GAGs) are linear polysaccharides and are among the primary components of mucosal surfaces in mammalian systems. The GAG layer lining the mucosal surface of the urinary tract is thought to play a critical role in urinary tract homeostasis and provide a barrier against urinary tract infection (UTI). This key component of the host-microbe interface may serve as a scaffolding site or a nutrient source for the urinary microbiota or invading pathogens, but its exact role in UTI pathogenesis is unclear. Although members of the gut microbiota have been shown to degrade GAGs, the utilization and degradation of GAGs by the urinary microbiota or uropathogens had not been investigated. In this study, we developed an in vitro plate-based assay to measure GAG degradation and utilization and used this assay to screen a library of 37 urinary bacterial isolates representing both urinary microbiota and uropathogenic species. This novel assay is more rapid, inexpensive, and quantitative compared to previously developed assays, and can measure three of the major classes of human GAGs. Our findings demonstrate that this assay captures the well-characterized ability of Streptococcus agalactiae to degrade hyaluronic acid and partially degrade chondroitin sulfate. Additionally, we present the first known report of chondroitin sulfate degradation by Proteus mirabilis, an important uropathogen and a causative agent of acute, recurrent, and catheter-associated urinary tract infections (CAUTI). In contrast, we observed that uropathogenic Escherichia coli (UPEC) and members of the urinary microbiota, including lactobacilli, were unable to degrade GAGs.
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spelling pubmed-87620502022-01-18 A Semi-Quantitative Assay to Measure Glycosaminoglycan Degradation by the Urinary Microbiota Nguyen, Vivian H. Khan, Fatima Shipman, Braden M. Neugent, Michael L. Hulyalkar, Neha V. Cha, Natalie Y. Zimmern, Philippe E. De Nisco, Nicole J. Front Cell Infect Microbiol Cellular and Infection Microbiology Glycosaminoglycans (GAGs) are linear polysaccharides and are among the primary components of mucosal surfaces in mammalian systems. The GAG layer lining the mucosal surface of the urinary tract is thought to play a critical role in urinary tract homeostasis and provide a barrier against urinary tract infection (UTI). This key component of the host-microbe interface may serve as a scaffolding site or a nutrient source for the urinary microbiota or invading pathogens, but its exact role in UTI pathogenesis is unclear. Although members of the gut microbiota have been shown to degrade GAGs, the utilization and degradation of GAGs by the urinary microbiota or uropathogens had not been investigated. In this study, we developed an in vitro plate-based assay to measure GAG degradation and utilization and used this assay to screen a library of 37 urinary bacterial isolates representing both urinary microbiota and uropathogenic species. This novel assay is more rapid, inexpensive, and quantitative compared to previously developed assays, and can measure three of the major classes of human GAGs. Our findings demonstrate that this assay captures the well-characterized ability of Streptococcus agalactiae to degrade hyaluronic acid and partially degrade chondroitin sulfate. Additionally, we present the first known report of chondroitin sulfate degradation by Proteus mirabilis, an important uropathogen and a causative agent of acute, recurrent, and catheter-associated urinary tract infections (CAUTI). In contrast, we observed that uropathogenic Escherichia coli (UPEC) and members of the urinary microbiota, including lactobacilli, were unable to degrade GAGs. Frontiers Media S.A. 2022-01-03 /pmc/articles/PMC8762050/ /pubmed/35047421 http://dx.doi.org/10.3389/fcimb.2021.803409 Text en Copyright © 2022 Nguyen, Khan, Shipman, Neugent, Hulyalkar, Cha, Zimmern and De Nisco https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Cellular and Infection Microbiology
Nguyen, Vivian H.
Khan, Fatima
Shipman, Braden M.
Neugent, Michael L.
Hulyalkar, Neha V.
Cha, Natalie Y.
Zimmern, Philippe E.
De Nisco, Nicole J.
A Semi-Quantitative Assay to Measure Glycosaminoglycan Degradation by the Urinary Microbiota
title A Semi-Quantitative Assay to Measure Glycosaminoglycan Degradation by the Urinary Microbiota
title_full A Semi-Quantitative Assay to Measure Glycosaminoglycan Degradation by the Urinary Microbiota
title_fullStr A Semi-Quantitative Assay to Measure Glycosaminoglycan Degradation by the Urinary Microbiota
title_full_unstemmed A Semi-Quantitative Assay to Measure Glycosaminoglycan Degradation by the Urinary Microbiota
title_short A Semi-Quantitative Assay to Measure Glycosaminoglycan Degradation by the Urinary Microbiota
title_sort semi-quantitative assay to measure glycosaminoglycan degradation by the urinary microbiota
topic Cellular and Infection Microbiology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8762050/
https://www.ncbi.nlm.nih.gov/pubmed/35047421
http://dx.doi.org/10.3389/fcimb.2021.803409
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