Author:
AA Kelani1, A Bruch1, C Visser2, AA Brakhage2, MG Blango1*
Author address:
1Junior Research Group RNA Biology of Fungal Infections, Leibniz Institute for Natural Product Research and Infection Biology (HKI), Jena, Germany
2Department of Molecular and Applied Microbiology, Leibniz Institute for Natural Product Research and Infection Biology (HKI), Jena, Germany
Full conference title:
10th Advances Against Aspergillosis and Mucormycosis
Date: 2 February 2022
Abstract:
Purpose:
Diagnostic markers and therapeutic options remain limited for identification and treatment of human fungal pathogens. RNA-based approaches offer a potential pathway forward, but many difficulties remain. As an example, previous study has shown that the cell wall of the filamentous fungus Aspergillus fumigatus is a cationic, impermeable barrier that limits uptake of many different drugs and sequesters negatively charged nucleic acids. Recent work revealed that host-derived extracellular vesicles produced by neutrophils are capable of delivering antifungal cargo across this cationic barrier to inhibit fungal growth, yet the RNA cargo of these vesicles remains unexplored. Our group aims to identify and characterize these extracellular RNAs with the hope of designing smarter therapeutic delivery options in the future. The protected RNA cargo of host-derived extracellular vesicles will likely also provide a window into the infection status of patients and improve early diagnosis.
Methods:
In the lab, we primarily study the pathogen A. fumigatus using a series of well-established molecular biological techniques and approaches. We then employ a mix of primary human immune cells and cell culture systems to investigate the contribution of RNA within extracellular vesicles to host defense against the pathogen. Extracellular vesicles are isolated using a variety of means, including centrifugation and size-exclusion chromatography.
Results:
In this study, we have isolated RNA from extracellular vesicles produced in response to fungal infection to identify the cargo delivered to the fungus during infection. Additionally, we have confirmed the presence of RNA binding proteins in extracellular vesicles that may contribute to loading specificity during infection. Finally, we have further characterized the small RNA machinery of A. fumigatus to better understand how a delivered RNA will be processed once internalized by the fungus and the parameters that dictate gene silencing in A. fumigatus.
Conclusion:
In the future, we hope that our increased understanding of the trafficking of RNA during fungal pathogenesis will facilitate the development of RNA-based diagnostics and therapeutics against human fungal pathogens and serve as a new way to combat these terrible infections.
Abstract Number: 64
Conference Year: 2022
Link to conference website: https://aaam2022.org/
URL Conference abstract: