MicroRNA-mediated calcineurin signaling activation induces CCL2, CCL3, CCL5, IL8, and chemotactic activities in 4,4′-methylene diphenyl diisocyanate exposed macrophages
Creators: name: Lin, Chen-Chung
name: Law, Brandon F.
name: Hettick, Justin M.
Publisher: name: Taylor & Francis
Year: 2021
Subjects: subject: Biochemistry
subject: Microbiology
subject: FOS: Biological sciences
subject: Cell Biology
subject: Molecular Biology
subject: Physiology
subject: Pharmacology
subject: 59999 Environmental Sciences not elsewhere classified
subject: FOS: Earth and related environmental sciences
subject: 39999 Chemical Sciences not elsewhere classified
subject: FOS: Chemical sciences
subject: Immunology
subject: FOS: Clinical medicine
subject: 69999 Biological Sciences not elsewhere classified
subject: Marine Biology
subject: Cancer
subject: Hematology
subject: 60506 Virology

description: Occupational exposure to 4,4′-methylene diphenyl diisocyanate (MDI), the most widely used monomeric diisocyanate, is one of the leading causes of occupational asthma (OA). Previously, we identified <i>microRNA (miR)-206-3p/miR-381-3p</i>-mediated PPP3CA/calcineurin signalling regulated <i>iNOS</i> transcription in macrophages and bronchoalveolar lavage cells (BALCs) after acute MDI exposure; however, whether PPP3CA/calcineurin signalling participates in regulation of other asthma-associated mediators secreted by macrophages/BALCs after MDI exposure is unknown.Several asthma-associated, macrophage-secreted mediator mRNAs from MDI exposed murine BALCs and MDI-glutathione (GSH) conjugate treated differentiated THP-1 macrophages were analysed using RT-qPCR.Endogenous <i>IL1B</i>, <i>TNF</i>, <i>CCL2</i>, <i>CCL3</i>, <i>CCL5</i>, and <i>TGFB1</i> were upregulated in MDI or MDI-GSH conjugate exposed BALCs and macrophages, respectively. Calcineurin inhibitor tacrolimus (FK506) attenuated the MDI-GSH conjugate-mediated induction of <i>CCL2</i>, <i>CCL3</i>, <i>CCL5</i>, and <i>CXCL8/IL8</i> but not others. Transfection of either miR-inhibitor-206-3p or miR-inhibitor-381-3p in macrophages induced chemokine <i>CCL2</i>, <i>CCL3</i>, <i>CCL5</i>, and <i>CXCL8</i> transcription, whereas FK506 attenuated the miR-inhibitor-206-3p or miR-inhibitor-381-3p-mediated effects. Finally, MDI-GSH conjugate treated macrophages showed increased chemotactic ability to various immune cells, which may be attenuated by FK506.In conclusion, these results indicate that MDI exposure to macrophages/BALCs may recruit immune cells into the airway via induction of chemokines by <i>miR-206-3p</i> and <i>miR-381-3p</i>-mediated calcineurin signalling activation. Occupational exposure to 4,4′-methylene diphenyl diisocyanate (MDI), the most widely used monomeric diisocyanate, is one of the leading causes of occupational asthma (OA). Previously, we identified <i>microRNA (miR)-206-3p/miR-381-3p</i>-mediated PPP3CA/calcineurin signalling regulated <i>iNOS</i> transcription in macrophages and bronchoalveolar lavage cells (BALCs) after acute MDI exposure; however, whether PPP3CA/calcineurin signalling participates in regulation of other asthma-associated mediators secreted by macrophages/BALCs after MDI exposure is unknown. Several asthma-associated, macrophage-secreted mediator mRNAs from MDI exposed murine BALCs and MDI-glutathione (GSH) conjugate treated differentiated THP-1 macrophages were analysed using RT-qPCR. Endogenous <i>IL1B</i>, <i>TNF</i>, <i>CCL2</i>, <i>CCL3</i>, <i>CCL5</i>, and <i>TGFB1</i> were upregulated in MDI or MDI-GSH conjugate exposed BALCs and macrophages, respectively. Calcineurin inhibitor tacrolimus (FK506) attenuated the MDI-GSH conjugate-mediated induction of <i>CCL2</i>, <i>CCL3</i>, <i>CCL5</i>, and <i>CXCL8/IL8</i> but not others. Transfection of either miR-inhibitor-206-3p or miR-inhibitor-381-3p in macrophages induced chemokine <i>CCL2</i>, <i>CCL3</i>, <i>CCL5</i>, and <i>CXCL8</i> transcription, whereas FK506 attenuated the miR-inhibitor-206-3p or miR-inhibitor-381-3p-mediated effects. Finally, MDI-GSH conjugate treated macrophages showed increased chemotactic ability to various immune cells, which may be attenuated by FK506. In conclusion, these results indicate that MDI exposure to macrophages/BALCs may recruit immune cells into the airway via induction of chemokines by <i>miR-206-3p</i> and <i>miR-381-3p</i>-mediated calcineurin signalling activation.