Abstract
Rhizopus delemar is an emerging fungal pathogen causing devastating mucormycosis in immunocompromised individuals. The organism remains understudied and there are urgent needs for new methods of rapid disease diagnosis for timely therapy. Extracellular vesicles with encapsulated RNAs have recently been discovered to have great potential applications for disease diagnoses and treatments. To explore the utilization of ex-RNA in studies of mucormycosis, we have performed RNA-Seq of ex-sRNAs from two clinical strains of R. delemar. Approximately 3.3 and 3.2 million clean reads were obtained from FGSC-9543 and CDC-8219 strains, respectively. The median sequence length of the sRNAs was 22 nts, with a minimum of 18 and a maximum of 30 nts. Further annotation identified 560 and 526 miRNAs from FGSC-9543 and CDC-8219 strains, respectively. miRNA target prediction and analysis of GO and KEGG pathways have revealed that the regulation of metabolism, secondary metabolite biosynthesis, and two-component system signaling are important during growth. We have also validated RNA-Seq by qRT-PCR and Northern blotting analysis of randomly selected miRNAs. Our results show that R. delemar has a rich reservoir of secreted ex-sRNAs and our studies could facilitate the development of improved diagnostic methods as well as elucidating virulence mechanisms for R. delemar infection.
Highlights
Mucormycosis is a highly invasive and life-threatening fungal infection that occurs in patients with immune deficiency due to hematologic diseases, diabetic ketoacidosis, or organ transplantation
Even though the numbers of exRNAs identified from C. albicans and C. neoformans were relatively low (2,000 and 1,244, respectively)[13,14], the results still suggested that R. delemar may produce extracellular vesicles (EVs) with encapsulated exRNAs
In our ongoing research effort to develop an efficient genetic transformation method for R. delemar, we were interested in the identification of type III RNA polymerase (Pol III) promoter sequences involved in the synthesis of a large variety of small non-coding RNA
Summary
Mucormycosis is a highly invasive and life-threatening fungal infection that occurs in patients with immune deficiency due to hematologic diseases, diabetic ketoacidosis, or organ transplantation. EVs and exRNAs have emerged as important mediators of intercellular communication that is involved in the regulation of a diverse range of biological processes. They are recognized in diseases, including cancer, neurodegenerative disorders, and infectious diseases, as they carry signals that identify themselves but are capable of altering the function of targeted cells (reviewed in[11,12]). Our studies provide a preliminary, yet important step in the development of exRNA signature for use in a biomarker-based diagnosis method for mucormycosis These exRNA profiles may illuminate molecular mechanisms associated with R. delemar pathogenesis within the host
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