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Author Spotlight: Accelerating Research on Bacterial Extracellular Vesicles Separation and Heterogeneity
Published on: September 1, 2023
The Origin of Plasma-Derived Bacterial Extracellular Vesicles in Healthy Individuals and Patients with Inflammatory
Emily Jones1, Régis Stentz1, Andrea Telatin1
1Gut Microbes and Health Research Programme, Quadram Institute, Norwich Research Park, Norwich NR4 7UQ, UK.
Abstract:
The gastrointestinal tract harbors the gut microbiota, structural alterations of which (dysbiosis) are linked with an increase in gut permeability ("leaky gut"), enabling luminal antigens and bacterial products such as nanosized bacterial extracellular vesicles (BEVs) to access the circulatory system. Blood-derived BEVs contain various cargoes and may be useful biomarkers for diagnosis and monitoring of disease status and relapse in conditions such as inflammatory bowel disease (IBD). To progress this concept, we developed a rapid, cost-effective protocol to isolate BEV-associated DNA and used 16S rRNA gene sequencing to identify bacterial origins of the blood microbiome of healthy individuals and patients with Crohn's disease and ulcerative colitis. The 16S rRNA gene sequencing successfully identified the origin of plasma-derived BEV DNA. The analysis showed that the blood microbiota richness, diversity, or composition in IBD, healthy control, and protocol control groups were not significantly distinct, highlighting the issue of 'kit-ome' contamination in low-biomass studies. Our pilot study provides the basis for undertaking larger studies to determine the potential use of blood microbiota profiling as a diagnostic aid in IBD.
Insights
Altered gut microbiota can increase gut permeability, allowing bacterial extracellular vesicles (BEVs) into the blood. This study explored blood BEV DNA for inflammatory bowel disease (IBD) diagnostics but found no significant differences due to contamination.
Area of Science:
- Microbiology
- Gastroenterology
- Immunology
Background:
- Gut dysbiosis is linked to increased intestinal permeability, allowing bacterial products like bacterial extracellular vesicles (BEVs) into circulation.
- Blood-borne BEVs may serve as biomarkers for inflammatory bowel disease (IBD) diagnosis and monitoring.
Purpose of the Study:
- To develop a method for isolating BEV-associated DNA from blood.
- To identify bacterial origins in the blood microbiome of healthy individuals and IBD patients using 16S rRNA gene sequencing.
- To assess the potential of blood microbiota profiling as a diagnostic aid for IBD.
Main Methods:
- Developed a rapid, cost-effective protocol for isolating BEV-associated DNA.
- Utilized 16S rRNA gene sequencing to analyze bacterial DNA from plasma-derived BEVs.
- Compared blood microbiota composition in IBD patients, healthy controls, and protocol controls.
Main Results:
- Successfully identified bacterial origins of plasma-derived BEV DNA.
- Found no significant differences in blood microbiota richness, diversity, or composition across groups.
- Identified 'kit-ome' contamination as a significant issue in low-biomass studies.
Conclusions:
- The developed protocol can identify bacterial origins of plasma-derived BEV DNA.
- Blood microbiota profiling showed no significant differences in IBD patients versus controls, likely due to contamination.
- Further large-scale studies are needed to validate blood microbiota profiling as an IBD diagnostic tool, addressing contamination issues.
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