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Published on: January 7, 2019
Dermatophagoides farinae microRNAs released to external environments via exosomes regulate inflammation-related gene
Kaiyue He1, Ting Yang2, Jinyan Yu3
1Department of Clinical Laboratory, Jiangnan University Medical Center, Wuxi, Jiangsu, China.
Background:
Dermatophagoides farinae (DFA) is an important species of house dust mites (HDMs) that causes allergic diseases. Previous studies have focused on allergens with protein components to explain the allergic effect of HDMs; however, there is little knowledge on the role of microRNAs (miRNAs) in the allergic effect of HDMs. This study aimed to unravel the new mechanism of dust mite sensitization from the perspective of cross-species transport of extracellular vesicles-encapsulated miRNAs from HDMs.
Methods:
Small RNA (sRNA) sequencing was performed to detect miRNAs expression profiles from DFA, DFA-derived exosomes and DFA culture supernatants. A quantitative fluorescent real-time PCR (qPCR) assay was used to detect miRNAs expression in dust specimens. BEAS-2B cells endocytosed exosomes were modeled in vitro to detect miRNAs from DFA and the expression of related inflammatory factors. Representative dfa-miR-276-3p and dfa-novel-miR2 were transfected into BEAS-2B cells, and then differentially expressed genes (DEGs) were analyzed by RNA sequencing. Protein-protein interaction (PPI) network analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway and Gene Ontology (GO) terms enrichment analyses were performed on the first 300 nodes of DEGs.
Results:
sRNA sequencing identified 42 conserved miRNAs and 66 novel miRNAs in DFA, DFA-derived exosomes, and DFA culture supernatants. A homology analysis was performed on the top 18 conserved miRNAs with high expression levels. The presence of dust mites and miRNAs from HDMs in living environment were also validated. Following uptake of DFA-derived exosomes by BEAS-2B cells, exosomes transported miRNAs from DFA to target cells and produced pro-inflammatory effects in corresponding cells. RNA sequencing identified DEGs in dfa-miR-276-3p and dfa-novel-miR2 transfected BEAS-2B cells. GO and KEGG enrichment analyses revealed the role of exosomes with cross-species transporting of DFA miRNAs in inflammatory signaling pathways, such as JAK-STAT signaling pathway, PI3K/AKT signaling pathway and IL-6-mediated signaling pathway.
Conclusion:
Our findings demonstrate the miRNAs expression profiles in DFA for the first time. The DFA miRNAs are delivered into living environments via exosomes, and engulfed by human bronchial epithelial cells, and cross-species regulation may contribute to inflammation-related processes.
Insights
House dust mites deliver microRNAs (miRNAs) via exosomes to human cells, triggering inflammation and allergic responses. This study reveals a novel cross-species mechanism contributing to dust mite sensitization and allergic diseases.
Area of Science:
- Molecular Biology
- Immunology
- Environmental Health
Background:
- House dust mites (HDMs), specifically *Dermatophagoides farinae* (DFA), are significant causes of allergic diseases.
- While protein allergens are well-studied, the role of microRNAs (miRNAs) in HDM-induced allergies remains largely unknown.
- This research investigates a novel mechanism of dust mite sensitization involving cross-species transfer of exosome-encapsulated miRNAs from HDMs.
Purpose of the Study:
- To profile microRNA (miRNA) expression in *Dermatophagoides farinae* (DFA) and its derived extracellular vesicles.
- To investigate the role of DFA-derived exosomal miRNAs in cross-species communication and induction of inflammatory responses in human cells.
- To elucidate the molecular pathways involved in dust mite sensitization mediated by miRNA transfer.
Main Methods:
- Small RNA sequencing to identify miRNAs in DFA, DFA-derived exosomes, and culture supernatants.
- Quantitative real-time PCR (qPCR) to detect miRNAs in dust specimens and assess exosome uptake by BEAS-2B cells.
- RNA sequencing and bioinformatics analyses (GO, KEGG, PPI) to identify differentially expressed genes and signaling pathways affected by transfected miRNAs.
Main Results:
- Identified 42 conserved and 66 novel miRNAs in DFA; confirmed presence of HDM miRNAs in environmental dust.
- Demonstrated that DFA-derived exosomes deliver miRNAs into human bronchial epithelial cells (BEAS-2B), inducing pro-inflammatory effects.
- RNA sequencing and pathway analyses revealed that transferred DFA miRNAs modulate key inflammatory pathways, including JAK-STAT and PI3K/AKT signaling.
Conclusions:
- This study provides the first comprehensive miRNA expression profile for DFA.
- DFA miRNAs are transported via exosomes into the environment and subsequently internalized by human cells.
- Cross-species miRNA regulation by dust mite exosomes represents a novel mechanism contributing to inflammation and allergic diseases.
