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Updated: May 10, 2026
Cerebrospinal Fluid MicroRNA Profiling Using Quantitative Real Time PCR
Published on: January 22, 2014
Microarray analysis of microRNA expression in peripheral blood mononuclear cells of critically ill patients with
1MOA Key Laboratory of Animal Biotechnology of National Ministry of Agriculture, Institute of Veterinary Immunology, and Research Laboratory of Virology, Immunology & Bioinformatics, Division of Veterinary Microbiology & Virology, Department of Preventive Veterinary Medicine, College of Veterinary Medicine, Northwest A & F University, Yangling, Xi'an City, Shaanxi Province, 712100, China.
Background:
With concerns about the disastrous health and economic consequences caused by the influenza pandemic, comprehensively understanding the global host response to influenza virus infection is urgent. The role of microRNA (miRNA) has recently been highlighted in pathogen-host interactions. However, the precise role of miRNAs in the pathogenesis of influenza virus infection in humans, especially in critically ill patients is still unclear.
Methods:
We identified cellular miRNAs involved in the host response to influenza virus infection by performing comprehensive miRNA profiling in peripheral blood mononuclear cells (PBMCs) from critically ill patients with swine-origin influenza pandemic H1N1 (2009) virus infection via miRNA microarray and quantitative reverse-transcription polymerase chain reaction (qRT-PCR) assays. Receiver operator characteristic (ROC) curve analysis was conducted and area under the ROC curve (AUC) was calculated to evaluate the diagnostic accuracy of severe H1N1 influenza virus infection. Furthermore, an integrative network of miRNA-mediated host-influenza virus protein interactions was constructed by integrating the predicted and validated miRNA-gene interaction data with influenza virus and host-protein-protein interaction information using Cytoscape software. Moreover, several hub genes in the network were selected and validated by qRT-PCR.
Results:
Forty-one significantly differentially expressed miRNAs were found by miRNA microarray; nine were selected and validated by qRT-PCR. QRT-PCR assay and ROC curve analyses revealed that miR-31, miR-29a and miR-148a all had significant potential diagnostic value for critically ill patients infected with H1N1 influenza virus, which yielded AUC of 0.9510, 0.8951 and 0.8811, respectively. We subsequently constructed an integrative network of miRNA-mediated host-influenza virus protein interactions, wherein we found that miRNAs are involved in regulating important pathways, such as mitogen-activated protein kinase signaling pathway, epidermal growth factor receptor signaling pathway, and Toll-like receptor signaling pathway, during influenza virus infection. Some of differentially expressed miRNAs via in silico analysis targeted mRNAs of several key genes in these pathways. The mRNA expression level of tumor protein T53 and transforming growth factor beta receptor 1 were found significantly reduced in critically ill patients, whereas the expression of Janus kinase 2, caspase 3 apoptosis-related cysteine peptidase, interleukin 10, and myxovirus resistance 1 were extremely increased in critically ill patients.
Conclusions:
Our data suggest that the dysregulation of miRNAs in the PBMCs of H1N1 critically ill patients can regulate a number of key genes in the major signaling pathways associated with influenza virus infection. These differentially expressed miRNAs could be potential therapeutic targets or biomarkers for severe influenza virus infection.
Insights
This study identifies specific microRNAs (miRNAs) as potential biomarkers for severe H1N1 influenza infection. These miRNAs, found in peripheral blood mononuclear cells (PBMCs), regulate key pathways and could offer new therapeutic targets for critical influenza cases.
Area of Science:
- Virology
- Immunology
- Genetics
Background:
- Influenza pandemics pose significant health and economic threats, necessitating a deeper understanding of host responses.
- MicroRNAs (miRNAs) play a role in pathogen-host interactions, but their specific involvement in severe human influenza, particularly in critically ill patients, remains unclear.
Purpose of the Study:
- To identify cellular miRNAs involved in the host response to influenza virus infection in critically ill patients.
- To evaluate the diagnostic accuracy of identified miRNAs for severe H1N1 influenza.
- To construct a miRNA-mediated host-pathogen interaction network.
Main Methods:
- Comprehensive miRNA profiling of peripheral blood mononuclear cells (PBMCs) from critically ill H1N1 patients using miRNA microarray and qRT-PCR.
- Receiver operator characteristic (ROC) curve analysis to assess diagnostic value.
- Integrative network construction of miRNA-gene and protein-protein interactions using Cytoscape.
Main Results:
- Forty-one differentially expressed miRNAs were identified; nine were validated by qRT-PCR.
- miR-31, miR-29a, and miR-148a showed significant diagnostic potential for severe H1N1 influenza (AUCs 0.9510, 0.8951, 0.8811).
- Network analysis revealed miRNA regulation of key pathways (MAPK, EGFR, Toll-like receptor) and identified altered expression of host genes (e.g., TP53, TGFBR1, JAK2, CASP3, IL10, MX1).
Conclusions:
- Dysregulated miRNAs in PBMCs of H1N1 patients modulate key genes in major signaling pathways relevant to influenza infection.
- These differentially expressed miRNAs represent potential therapeutic targets and biomarkers for severe influenza virus infection.
