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Exosomal Micro RNA Isolation in White-Tailed Deer (Odocoileus virginianus) for Diagnostic Biomarker Discovery
Maite De María1, Lillian G Maxwell2,3, Margaret E Hunter2
1Cherokee Nation System Solutions, contractor to the U.S. Geological Survey Wetland and Aquatic Research Center, 7920 NW 71st Street, Gainesville, Florida 32653, USA.
Abstract:
Molecular approaches are becoming more prevalent for the diagnosis of neurodegenerative diseases in human medicine and can be extended to diagnosis of wildlife diseases such as chronic wasting disease and other prion diseases. These diseases have been associated with exosome-bound molecular biomarkers of disease progression, such as proteins and micro RNA molecules (miRNA). We tested and optimized a method for exosomal miRNA isolation from minimally invasive, small-volume serum samples obtained from white-tailed deer (Odocoileus virginianus). We confirmed the isolation of exosomes and optimized a commercially available benchtop kit to obtain sufficient and pure RNA for miRNA sequencing. The selected method for RNA extraction combines two 500-μL serum aliquots into one elution column and re-eluting the final product of the column. We identified 137 miRNA present in healthy white-tailed deer that can be used as a baseline to identify putative miRNA biomarkers of disease progression and mechanisms of infection in future comparative disease studies. This approach to biomarker discovery may help to inform biological processes in wildlife populations and provide alternatives to invasive or postmortem samples.
Insights
Researchers optimized a method to isolate exosomal microRNA (miRNA) from white-tailed deer serum. This technique identifies potential miRNA biomarkers for wildlife prion diseases, offering a less invasive diagnostic approach.
Area of Science:
- Veterinary Medicine
- Molecular Biology
- Wildlife Disease Diagnostics
Background:
- Neurodegenerative diseases like chronic wasting disease (CWD) are increasingly diagnosed using molecular biomarkers.
- Exosomes carrying microRNA (miRNA) are associated with disease progression in prion diseases.
- Developing non-invasive diagnostic methods for wildlife is crucial.
Purpose of the Study:
- To optimize a method for isolating exosomal miRNA from small-volume white-tailed deer serum samples.
- To establish a baseline of miRNA profiles in healthy deer for future biomarker discovery.
- To support the development of molecular diagnostics for wildlife prion diseases.
Main Methods:
- Serum samples from white-tailed deer (Odocoileus virginianus) were used.
- A commercially available kit was optimized for exosomal RNA extraction, combining two 500-μL aliquots.
- Exosome isolation and RNA purity for miRNA sequencing were confirmed.
Main Results:
- A reliable method for exosomal miRNA isolation from deer serum was established and optimized.
- 137 miRNAs were identified in healthy white-tailed deer, providing a baseline dataset.
- Sufficient pure RNA was obtained for miRNA sequencing.
Conclusions:
- The optimized method enables the discovery of exosomal miRNA biomarkers in wildlife.
- This approach offers a less invasive alternative to traditional diagnostic methods for prion diseases.
- The identified baseline miRNA profile aids in understanding disease mechanisms and progression in wildlife populations.
