SEX INFLUENCES ON THE GUT MICROBIOME IN A MOUSE MODEL OF AN ENVIRONMENTAL ALLERGEN CHALLENGE
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Date
2023-05
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[Bloomington, Ind.] : Indiana University
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Abstract
Asthma is a chronic inflammatory disease of the airway that compromises lung function and affects millions of people worldwide. Disparities in sensitivity between men and women to the initiation and exacerbation of asthma phenotypes have been identified; however, it remains unclear whether mediators such as the gut microbiome contribute to such differences. Commensal microbial communities found throughout the body are necessary for initiating immune responses and they contribute to the development of chronic inflammation. In this study, we used a mouse model of house dust mite (HDM) challenge and identified sex differences in the composition of the gut and lung microbiota. The aim being to discover to what extent the microbiome undergoes alteration in response to the induction of allergic asthma in mice challenged with HDM. For this, fecal pellets and whole lung tissue of male and female C57BL/6 mice intranasally exposed to 25 µg of house dust mite extract (HDM) in 50µL of phosphate buffered saline (PBS) or 50µL PBS (control) daily for 5 weeks (n=4-6/ treatment group) were collected before starting treatment and at the end of week 5. DNA from fecal pellets was extracted using the ZymoBIOMICS®-96 MagBead DNA Kit and analyzed with the ZymoBIOMICS® Service to determine the 16S microbiome: Targeted Metagenomic Sequencing (Zymo Research, Irvine, CA). DNA isolation from whole lung tissue and 16S rRNA sequencing was performed by the Microbiome Core Facility of the University of North Carolina at Chapel Hill. Visual and statistical comparisons using stacked taxa bar plots and Firmicutes to Bacteroidetes ratios expressed community composition changes in samples. Nonmetric Multidimensional Scaling illustrated structural alteration to the microbiota, which was verified through PERMANOVA testing. We concluded that the gut microbiome did experience statistically significant sex-specific changes to its composition and structure; while the lung microbiome saw significant differences as a result of treatment, not sex.
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Thesis (Master of Science) - Indiana University, Department of Environmental and Occupational Health of the School of Public Health, 2023
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microbiome, sex differences, asthma, environmental exposure
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Thesis