MiR-1165-3p Suppresses Th2 Differentiation via Targeting IL-13 and PPM1A in a Mouse Model of Allergic Airway Inflammation

Abstract

Purpose CD4 + T cells are essential in the pathogenesis of allergic asthma. We have previously demonstrated that microRNA-1165-3p (miR-1165-3p) was significantly reduced in T-helper type (Th) 2 cells and that miR-1165-3p was a surrogate marker for atopic asthma. Little is known about the mechanisms of miR-1165-3p in the regulation of Th2-dominated allergic inflammation. We aimed to investigate the associations between Th2 differentiation and miR-1165b-3p in asthma as well as the possible mechanisms. Methods CD4 + naïve T cells were differentiated into Th1 or Th2 cells in vitro . MiR-1165-3p was up-regulated or down-regulated using lentiviral systems during Th1/Th2 differentiation. In vivo , the lentiviral particles with the miR-1165-3p enhancer were administered by tail vein injection on the first day of a house dust mite -induced allergic airway inflammation model. Allergic inflammation and Th1/Th2 differentiation were routinely monitored. To investigate the potential targets of miR-1165-3p, biotin-microRNA pull-down products were sequenced, and the candidates were further verified with a dual-luciferase reporter assay. The roles of a target protein phosphatase, Mg 2+ /Mn 2+ -dependent 1A (PPM1A), in Th2 cell differentiation and allergic asthma were further explored. Plasma PPM1A was determined by ELISA in 18 subjects with asthma and 20 controls. Results The lentivirus encoding miR-1165-3p suppressed Th2-cell differentiation in vitro . In contrast, miR-1165-3p silencing promoted Th2-cell development. In the HDM-induced model of allergic airway inflammation, miR-1165-3p up-regulation was accompanied by reduced airway hyper-responsiveness, serum immunoglobulin E, airway inflammation and Th2-cell polarization. IL-13 and PPM1A were the direct targets of miR-1165-3p. The expression of IL-13 or PPM1A was inversely correlated with that of miR-1165-3p. PPM1A regulated the signal transducer and activator of transcription and AKT signaling pathways during Th2 differentiation. Moreover, plasma PPM1A was significantly increased in asthmatic patients. Conclusions MiR-1165-3p negatively may regulate Th2-cell differentiation by targeting IL-13 and PPM1A in allergic airway inflammation.

A comprehensive analysis of microRNAs as diagnostic biomarkers for asthma

“…Furthermore, the upregulation of miR-1165-3p could lead to a decrease in airway hyperreactivity and airway inflammation through directly targeting IL-13. 119 As for miR-185-5p, it participates in calcium signaling by targeting NFAT and CaMKII proteins in cardiomyocytes, and may play a role in muscle cell hypertrophy, proliferation, and cell contraction in asthma. 120 , 121 These results suggest that these biomarker candidates play a role in the pathogenesis of asthma.…”

KIF2A decreases IL-33 production and attenuates allergic asthmatic inflammation

“…Total RNA was isolated from frozen tissues or cells using TRIzol (Invitrogen), and cDNA was synthesized using 5X All-In-One RT MasterMix (G490, Abm, Zhenjiangg China) according to the manufacturer’s instructions as described previously [ 20 ]. qPCR analysis was performed using a StepOnePlus Real-Time PCR System (ABI, USA) in conjunction with SYBR Advantage qPCR Premix (63976, TaKaRa Bio).…”

Epigenetic Regulation of Th2 Response in Asthma by Non-Coding RNAs

“…A study has shown that miR-1165-3p targets IL-13 and PPM1A to control Th2 differentiation and pulmonary inflammation in asthma. miR-1165-3p inhibits the Th2 response of allergy through the STAT and AKT signaling pathways by targeted inhibition of protein phosphatase, Mg 2+/Mn 2+ − dependent 1A (PPM1A), thus proving that miR-1165-3p and PPM1A may be effective targets for the prevention and treatment of allergic asthma and related diseases [41]. The miR-29c/B7-H3 axis plays an important role in asthmatic children by regulating Th2/Th17 cell differentiation, and may provide a new target for asthma treatment [42].…”