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MicroRNA regulation of human protease genes essential for influenza virus replication.

Meliopoulos VA, Andersen LE, Brooks P, Yan X, Bakre A, Coleman JK, Tompkins SM, Tripp RA - PLoS ONE (2012)

Bottom Line: However, the rapid emergence of drug resistance has emphasized the need for new drug targets.The genes validated as critical for influenza virus replication were ADAMTS7, CPE, DPP3, MST1, and PRSS12, and pathway analysis showed these genes were in global host cell pathways governing inflammation (NF-κB), cAMP/calcium signaling (CRE/CREB), and apoptosis.Analyses of host microRNAs predicted to govern expression of these genes showed that eight miRNAs regulated gene expression during virus replication.

View Article: PubMed Central - PubMed

Affiliation: Department of Infectious Diseases, University of Georgia, Athens, Georgia, United States of America.

ABSTRACT
Influenza A virus causes seasonal epidemics and periodic pandemics threatening the health of millions of people each year. Vaccination is an effective strategy for reducing morbidity and mortality, and in the absence of drug resistance, the efficacy of chemoprophylaxis is comparable to that of vaccines. However, the rapid emergence of drug resistance has emphasized the need for new drug targets. Knowledge of the host cell components required for influenza replication has been an area targeted for disease intervention. In this study, the human protease genes required for influenza virus replication were determined and validated using RNA interference approaches. The genes validated as critical for influenza virus replication were ADAMTS7, CPE, DPP3, MST1, and PRSS12, and pathway analysis showed these genes were in global host cell pathways governing inflammation (NF-κB), cAMP/calcium signaling (CRE/CREB), and apoptosis. Analyses of host microRNAs predicted to govern expression of these genes showed that eight miRNAs regulated gene expression during virus replication. These findings identify unique host genes and microRNAs important for influenza replication providing potential new targets for disease intervention strategies.

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Related in: MedlinePlus

Effect of miRNA inhibition on influenza replication.A549 cells were treated with the appropriate miRNA inhibitor (25 nM) for 48 hours, followed by infection with A/WSN/33 (MOI = 0.001). Cellular supernatant was tested for infectious virus production by a modified TCID50 48 hpi. Data is expressed as TCID50/ml and is representative of two independent experiments.
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pone-0037169-g006: Effect of miRNA inhibition on influenza replication.A549 cells were treated with the appropriate miRNA inhibitor (25 nM) for 48 hours, followed by infection with A/WSN/33 (MOI = 0.001). Cellular supernatant was tested for infectious virus production by a modified TCID50 48 hpi. Data is expressed as TCID50/ml and is representative of two independent experiments.

Mentions: Given the evidence that miR-1254, miR-1272, miR-17-5p, miR-17-3p, miR-106B, miR-106B*, miR-124-a, and miR-124* are involved in governing aspects of ADAMTS7, CPE, DPP3, MST1, and PRSS12 gene expression (Figure 5), the role of these miRNAs in the regulation of influenza virus replication was determined (Figure 6). A549 cells were treated with individual miRNA inhibitors and the cells infected with A/WSN/33 to determine the effect on virus replication (Figure 6). Of the 8 miRNA inhibitors tested, inhibition of miR-106B was associated with a decrease in influenza virus replication, while inhibition of miR-124 resulted in an increase in virus replication with respect to the negative control. Inhibition of the other eight miRNAs had more subtle effects with slight increases or decreases of influenza virus replication. The decrease of virus replication associated with inhibition of miR-106B is likely associated with a decrease of CPE gene expression as RNAi silencing of CPE was associated with low levels of virus replication (Figures 1 and 2). The level of virus replication was confirmed by qPCR M gene levels and was consistent with the findings observed in Figure 5 (data not shown). The results show that some of the miRNAs modulate host genes critical for influenza virus replication, thus it likely that the tempo of host gene expression is differentially regulated in response to influenza virus infection. Further, the results provide evidence that targeting miRNAs may offer an alternative disease intervention approach to control influenza virus replication.


MicroRNA regulation of human protease genes essential for influenza virus replication.

Meliopoulos VA, Andersen LE, Brooks P, Yan X, Bakre A, Coleman JK, Tompkins SM, Tripp RA - PLoS ONE (2012)

Effect of miRNA inhibition on influenza replication.A549 cells were treated with the appropriate miRNA inhibitor (25 nM) for 48 hours, followed by infection with A/WSN/33 (MOI = 0.001). Cellular supernatant was tested for infectious virus production by a modified TCID50 48 hpi. Data is expressed as TCID50/ml and is representative of two independent experiments.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3351457&req=5

pone-0037169-g006: Effect of miRNA inhibition on influenza replication.A549 cells were treated with the appropriate miRNA inhibitor (25 nM) for 48 hours, followed by infection with A/WSN/33 (MOI = 0.001). Cellular supernatant was tested for infectious virus production by a modified TCID50 48 hpi. Data is expressed as TCID50/ml and is representative of two independent experiments.
Mentions: Given the evidence that miR-1254, miR-1272, miR-17-5p, miR-17-3p, miR-106B, miR-106B*, miR-124-a, and miR-124* are involved in governing aspects of ADAMTS7, CPE, DPP3, MST1, and PRSS12 gene expression (Figure 5), the role of these miRNAs in the regulation of influenza virus replication was determined (Figure 6). A549 cells were treated with individual miRNA inhibitors and the cells infected with A/WSN/33 to determine the effect on virus replication (Figure 6). Of the 8 miRNA inhibitors tested, inhibition of miR-106B was associated with a decrease in influenza virus replication, while inhibition of miR-124 resulted in an increase in virus replication with respect to the negative control. Inhibition of the other eight miRNAs had more subtle effects with slight increases or decreases of influenza virus replication. The decrease of virus replication associated with inhibition of miR-106B is likely associated with a decrease of CPE gene expression as RNAi silencing of CPE was associated with low levels of virus replication (Figures 1 and 2). The level of virus replication was confirmed by qPCR M gene levels and was consistent with the findings observed in Figure 5 (data not shown). The results show that some of the miRNAs modulate host genes critical for influenza virus replication, thus it likely that the tempo of host gene expression is differentially regulated in response to influenza virus infection. Further, the results provide evidence that targeting miRNAs may offer an alternative disease intervention approach to control influenza virus replication.

Bottom Line: However, the rapid emergence of drug resistance has emphasized the need for new drug targets.The genes validated as critical for influenza virus replication were ADAMTS7, CPE, DPP3, MST1, and PRSS12, and pathway analysis showed these genes were in global host cell pathways governing inflammation (NF-κB), cAMP/calcium signaling (CRE/CREB), and apoptosis.Analyses of host microRNAs predicted to govern expression of these genes showed that eight miRNAs regulated gene expression during virus replication.

View Article: PubMed Central - PubMed

Affiliation: Department of Infectious Diseases, University of Georgia, Athens, Georgia, United States of America.

ABSTRACT
Influenza A virus causes seasonal epidemics and periodic pandemics threatening the health of millions of people each year. Vaccination is an effective strategy for reducing morbidity and mortality, and in the absence of drug resistance, the efficacy of chemoprophylaxis is comparable to that of vaccines. However, the rapid emergence of drug resistance has emphasized the need for new drug targets. Knowledge of the host cell components required for influenza replication has been an area targeted for disease intervention. In this study, the human protease genes required for influenza virus replication were determined and validated using RNA interference approaches. The genes validated as critical for influenza virus replication were ADAMTS7, CPE, DPP3, MST1, and PRSS12, and pathway analysis showed these genes were in global host cell pathways governing inflammation (NF-κB), cAMP/calcium signaling (CRE/CREB), and apoptosis. Analyses of host microRNAs predicted to govern expression of these genes showed that eight miRNAs regulated gene expression during virus replication. These findings identify unique host genes and microRNAs important for influenza replication providing potential new targets for disease intervention strategies.

Show MeSH
Related in: MedlinePlus