Limits...
Integrated microRNA-mRNA-analysis of human monocyte derived macrophages upon Mycobacterium avium subsp. hominissuis infection.

Sharbati J, Lewin A, Kutz-Lohroff B, Kamal E, Einspanier R, Sharbati S - PLoS ONE (2011)

Bottom Line: We furthermore outlined a regulatory network of potential interactions between microRNAs and mRNAs.This study provides a theoretical concept for unveiling how distinct mycobacteria could manipulate host cell response.In addition, functional relevance was confirmed by uncovering the control of major caspases 3 and 7 by let-7e and miR-29a, respectively.

View Article: PubMed Central - PubMed

Affiliation: Institute of Veterinary Biochemistry, Freie Universitaet Berlin, Berlin, Germany. sharbati@zedat.fu-berlin.de

ABSTRACT

Background: Many efforts have been made to understand basal mechanisms of mycobacterial infections. Macrophages are the first line of host immune defence to encounter and eradicate mycobacteria. Pathogenic species have evolved different mechanisms to evade host response, e.g. by influencing macrophage apoptotic pathways. However, the underlying molecular regulation is not fully understood. A new layer of eukaryotic regulation of gene expression is constituted by microRNAs. Therefore, we present a comprehensive study for identification of these key regulators and their targets in the context of host macrophage response to mycobacterial infections.

Methodology/principal findings: We performed microRNA as well as mRNA expression analysis of human monocyte derived macrophages infected with several Mycobacterium avium hominissuis strains by means of microarrays as well as quantitative reverse transcription PCR (qRT-PCR). The data revealed the ability of all strains to inhibit apoptosis by transcriptional regulation of BCL2 family members. Accordingly, at 48 h after infection macrophages infected with all M. avium strains showed significantly decreased caspase 3 and 7 activities compared to the controls. Expression of let-7e, miR-29a and miR-886-5p were increased in response to mycobacterial infection at 48 h. The integrated analysis of microRNA and mRNA expression as well as target prediction pointed out regulative networks identifying caspase 3 and 7 as potential targets of let-7e and miR-29a, respectively. Consecutive reporter assays verified the regulation of caspase 3 and 7 by these microRNAs.

Conclusions/significance: We show for the first time that mycobacterial infection of human macrophages causes a specific microRNA response. We furthermore outlined a regulatory network of potential interactions between microRNAs and mRNAs. This study provides a theoretical concept for unveiling how distinct mycobacteria could manipulate host cell response. In addition, functional relevance was confirmed by uncovering the control of major caspases 3 and 7 by let-7e and miR-29a, respectively.

Show MeSH

Related in: MedlinePlus

Regulatory network of MAH infected MDMs deduced from integrated analysis of miRNA-mRNA microarray data.Negatively correlated miRNA-mRNA interactions were visualised as a network using Cytoscape. This network gives for the first time an theoretical outline of the concerted action of regulating miRNAs (blue triangles) and their potential target mRNAs (green circles) in mycobacterial infection of human macrophages.
© Copyright Policy
Related In: Results  -  Collection


getmorefigures.php?uid=PMC3101234&req=5

pone-0020258-g002: Regulatory network of MAH infected MDMs deduced from integrated analysis of miRNA-mRNA microarray data.Negatively correlated miRNA-mRNA interactions were visualised as a network using Cytoscape. This network gives for the first time an theoretical outline of the concerted action of regulating miRNAs (blue triangles) and their potential target mRNAs (green circles) in mycobacterial infection of human macrophages.

Mentions: As described in the previous section, our data comprised many mRNAs and miRNAs possessing a negatively correlated expression. Some of these correlations are validated and published interactions between miRNAs and their target mRNAs, however many potential interactions are still to be discovered. Our aim was to create a theoretical concept of an interacting network based on linking miRNA and mRNA expression data to target prediction. To facilitate the prediction of possible interactions from comprehensive data presented here, we have employed the integrative analysis of target prediction as well as miRNA and mRNA expression by using the tool MAGIA [19]. We first predicted targets of all detected miRNAs using the combined PITA and Target Scan analysis implemented in the MAGIA query (not considering the expression data). The hereby identified target list was initially subjected to pathway analysis using the Cytoscape Plug-In ClueGO (table S1). It was intriguing to find out that predicted targets of miRNAs were enriched in pathways being mainly involved in cell death/apoptosis and immune signalling. This very well reflected affected cellular processes determined by mRNA expression analysis as described above (table 2). Subsequently, we performed the MAGIA analysis considering the expression data of dysregulated mRNAs showing at least 2-fold altered expression or a temporal expression pattern (figure 1 A–C) and considering all detected miRNAs of corresponding samples, respectively. Table S2 summarises miRNAs and their predicted target genes including the calculated correlation coefficients (Pearson r). Negatively correlated miRNA-mRNA interactions were visualised as a network using Cytoscape (figure 2). This network gives for the first time a theoretical outline of the concerted action of regulating miRNAs and their potential target mRNAs in mycobacterial infection of macrophages.


Integrated microRNA-mRNA-analysis of human monocyte derived macrophages upon Mycobacterium avium subsp. hominissuis infection.

Sharbati J, Lewin A, Kutz-Lohroff B, Kamal E, Einspanier R, Sharbati S - PLoS ONE (2011)

Regulatory network of MAH infected MDMs deduced from integrated analysis of miRNA-mRNA microarray data.Negatively correlated miRNA-mRNA interactions were visualised as a network using Cytoscape. This network gives for the first time an theoretical outline of the concerted action of regulating miRNAs (blue triangles) and their potential target mRNAs (green circles) in mycobacterial infection of human macrophages.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3101234&req=5

pone-0020258-g002: Regulatory network of MAH infected MDMs deduced from integrated analysis of miRNA-mRNA microarray data.Negatively correlated miRNA-mRNA interactions were visualised as a network using Cytoscape. This network gives for the first time an theoretical outline of the concerted action of regulating miRNAs (blue triangles) and their potential target mRNAs (green circles) in mycobacterial infection of human macrophages.
Mentions: As described in the previous section, our data comprised many mRNAs and miRNAs possessing a negatively correlated expression. Some of these correlations are validated and published interactions between miRNAs and their target mRNAs, however many potential interactions are still to be discovered. Our aim was to create a theoretical concept of an interacting network based on linking miRNA and mRNA expression data to target prediction. To facilitate the prediction of possible interactions from comprehensive data presented here, we have employed the integrative analysis of target prediction as well as miRNA and mRNA expression by using the tool MAGIA [19]. We first predicted targets of all detected miRNAs using the combined PITA and Target Scan analysis implemented in the MAGIA query (not considering the expression data). The hereby identified target list was initially subjected to pathway analysis using the Cytoscape Plug-In ClueGO (table S1). It was intriguing to find out that predicted targets of miRNAs were enriched in pathways being mainly involved in cell death/apoptosis and immune signalling. This very well reflected affected cellular processes determined by mRNA expression analysis as described above (table 2). Subsequently, we performed the MAGIA analysis considering the expression data of dysregulated mRNAs showing at least 2-fold altered expression or a temporal expression pattern (figure 1 A–C) and considering all detected miRNAs of corresponding samples, respectively. Table S2 summarises miRNAs and their predicted target genes including the calculated correlation coefficients (Pearson r). Negatively correlated miRNA-mRNA interactions were visualised as a network using Cytoscape (figure 2). This network gives for the first time a theoretical outline of the concerted action of regulating miRNAs and their potential target mRNAs in mycobacterial infection of macrophages.

Bottom Line: We furthermore outlined a regulatory network of potential interactions between microRNAs and mRNAs.This study provides a theoretical concept for unveiling how distinct mycobacteria could manipulate host cell response.In addition, functional relevance was confirmed by uncovering the control of major caspases 3 and 7 by let-7e and miR-29a, respectively.

View Article: PubMed Central - PubMed

Affiliation: Institute of Veterinary Biochemistry, Freie Universitaet Berlin, Berlin, Germany. sharbati@zedat.fu-berlin.de

ABSTRACT

Background: Many efforts have been made to understand basal mechanisms of mycobacterial infections. Macrophages are the first line of host immune defence to encounter and eradicate mycobacteria. Pathogenic species have evolved different mechanisms to evade host response, e.g. by influencing macrophage apoptotic pathways. However, the underlying molecular regulation is not fully understood. A new layer of eukaryotic regulation of gene expression is constituted by microRNAs. Therefore, we present a comprehensive study for identification of these key regulators and their targets in the context of host macrophage response to mycobacterial infections.

Methodology/principal findings: We performed microRNA as well as mRNA expression analysis of human monocyte derived macrophages infected with several Mycobacterium avium hominissuis strains by means of microarrays as well as quantitative reverse transcription PCR (qRT-PCR). The data revealed the ability of all strains to inhibit apoptosis by transcriptional regulation of BCL2 family members. Accordingly, at 48 h after infection macrophages infected with all M. avium strains showed significantly decreased caspase 3 and 7 activities compared to the controls. Expression of let-7e, miR-29a and miR-886-5p were increased in response to mycobacterial infection at 48 h. The integrated analysis of microRNA and mRNA expression as well as target prediction pointed out regulative networks identifying caspase 3 and 7 as potential targets of let-7e and miR-29a, respectively. Consecutive reporter assays verified the regulation of caspase 3 and 7 by these microRNAs.

Conclusions/significance: We show for the first time that mycobacterial infection of human macrophages causes a specific microRNA response. We furthermore outlined a regulatory network of potential interactions between microRNAs and mRNAs. This study provides a theoretical concept for unveiling how distinct mycobacteria could manipulate host cell response. In addition, functional relevance was confirmed by uncovering the control of major caspases 3 and 7 by let-7e and miR-29a, respectively.

Show MeSH
Related in: MedlinePlus