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Published on: May 31, 2018
Targeting of microRNA-142-3p in dendritic cells regulates endotoxin-induced mortality
Yaping Sun1, Sooryanarayana Varambally, Christopher A Maher
1Department of Internal Medicine, University of Michigan Comprehensive Cancer Center, Ann Arbor, MI, USA.
Abstract:
While miRNAs are increasingly linked to various immune responses, whether they can be targeted for regulating in vivo inflammatory processes such as endotoxin-induced Gram-negative sepsis is not known. Production of cytokines by the dendritic cells (DCs) plays a critical role in response to endotoxin, lipopolysaccharide (LPS). We profiled the miRNA and mRNA of CD11c⁺ DCs in an unbiased manner and found that at baseline, miR-142-3p was among the most highly expressed endogenous miRs while IL-6 was among the most highly expressed mRNA after LPS stimulation. Multiple computational algorithms predicted the IL-6 3' untranslated region (UTR) to be a target of miR-142-3p. Studies using luciferase reporters carrying wild-type (WT) and mutant IL-6 3'UTR confirmed IL-6 as a target for miR-142-3p. In vitro knockdown and overexpression studies demonstrated a critical and specific role for miR142-3p in regulating IL-6 production by the DCs after LPS stimulation. Importantly, treatment of only WT but not the IL-6-deficient (IL-6(⁻/⁻)) mice with locked nucleic acid (LNA)-modified phosphorothioate oligonucleotide complementary to miR 142-3p reduced endotoxin-induced mortality. These results demonstrate a critical role for miR-142-3p in regulating DC responses to LPS and provide proof of concept for targeting miRs as a novel strategy for treatment of endotoxin-induced mortality.
Insights
MicroRNAs (miRNAs) can regulate immune responses. Targeting miR-142-3p in dendritic cells (DCs) reduced mortality in endotoxin-induced sepsis, suggesting a novel therapeutic strategy.
Area of Science:
- Immunology
- Molecular Biology
- Genetics
Background:
- MicroRNAs (miRNAs) are implicated in immune responses.
- The role of miRNAs in regulating endotoxin-induced inflammatory processes, like Gram-negative sepsis, remains unclear.
- Dendritic cells (DCs) and their cytokine production are critical in responding to lipopolysaccharide (LPS).
Purpose of the Study:
- To investigate the role of miRNAs in regulating DC responses to LPS.
- To determine if targeting miRNAs can be a therapeutic strategy for endotoxin-induced mortality.
Main Methods:
- miRNA and mRNA profiling of CD11c+ DCs after LPS stimulation.
- Computational prediction and luciferase reporter assays to identify miRNA targets.
- In vitro knockdown and overexpression studies of miR-142-3p in DCs.
- In vivo studies using wild-type and IL-6-deficient mice treated with miR-142-3p inhibitors.
Main Results:
- miR-142-3p was highly expressed in DCs and predicted to target IL-6 mRNA.
- miR-142-3p specifically regulated IL-6 production by DCs in response to LPS.
- In vivo administration of a miR-142-3p inhibitor reduced mortality in wild-type mice with LPS-induced sepsis but not in IL-6-deficient mice.
Conclusions:
- miR-142-3p plays a critical role in regulating DC responses to LPS.
- Targeting miR-142-3p offers a potential therapeutic approach for endotoxin-induced mortality and sepsis.
