miR-142 controls metabolic reprogramming that regulates dendritic cell activation
Yaping Sun, … , Thomas Saunders, Pavan Reddy
Yaping Sun, … , Thomas Saunders, Pavan Reddy
Published April 8, 2019
Citation Information: J Clin Invest. 2019;129(5):2029-2042. https://doi.org/10.1172/JCI123839.
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Research Article Hematology Immunology Article has an altmetric score of 4

miR-142 controls metabolic reprogramming that regulates dendritic cell activation

Abstract

DCs undergo metabolic reprogramming from a predominantly oxidative phosphorylation (OXPHOS) to glycolysis to mount an immunogenic response. The mechanism underpinning the metabolic reprogramming remains elusive. We demonstrate that miRNA-142 (miR-142) is pivotal for this shift in metabolism, which regulates the tolerogenic and immunogenic responses of DCs. In the absence of miR-142, DCs fail to switch from OXPHOS and show reduced production of proinflammatory cytokines and the ability to activate T cells in vitro and in in vivo models of sepsis and alloimmunity. Mechanistic studies demonstrate that miR-142 regulates fatty acid (FA) oxidation, which causes the failure to switch to glycolysis. Loss- and gain-of-function experiments identified carnitine palmitoyltransferase -1a (CPT1a), a key regulator of the FA pathway, as a direct target of miR-142 that is pivotal for the metabolic switch. Thus, our findings show that miR-142 is central to the metabolic reprogramming that specifically favors glycolysis and immunogenic response by DCs.

Authors

Yaping Sun, Katherine Oravecz-Wilson, Sydney Bridges, Richard McEachin, Julia Wu, Stephanie H. Kim, Austin Taylor, Cynthia Zajac, Hideaki Fujiwara, Daniel Christopher Peltier, Thomas Saunders, Pavan Reddy

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Figure 6

miR-142 targets CPT1a and regulates FAO in DCs.

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miR-142 targets CPT1a and regulates FAO in DCs. (A) Enrichment score of ...
(A) Enrichment score of gene set for FAO pathway in WT and miR-142–/– DCs (biological triplicates) analysis by GSEA. (B) WT and miR-142–/– DCs were treated with LPS for indicated times. Cell lysates were processed for immunoblotting using Abs against CPT1a and β-actin. A representative blot is shown (top), and densitometry data (bottom) from 3 independent experiments were combined (mean ± SEM). (C) WT and miR-142–/– DCs treated with LPS for indicated times and processed for qPCR using specific primers for CPT1a. Data from 3 similar experiments were combined (mean ± SEM). (D) WT and miR-142–/– DCs were treated with LPS for indicated times, then processed for FAO assay as in Methods. CPM values were collected from biological quadruplicate or quintuplet samples. Data from 2 similar experiments were combined (mean ± SEM). (E) miR-142–/– DCs were transfected with a pool of CPT1a siRNAs or scramble control as described in Methods. The whole lysates were processed for immunoblotting against CPT1a and β-actin. A representative blot is shown (top), as are densitometry data combined from 3 independent experiments (mean ± SEM). (F) WT and miR-142–/– DCs transfected with lentiviruses, treated with LPS for indicated times, then processed for FAO assay in quadruplicates or quintuplets as described in Methods. Data are combined from 2 independent experiments (mean ± SEM). (G) WT and miR-142–/– DCs were treated with LPS for 12 hours, then treated with etomoxir or oligomicin. OCR values were recorded by Seahorse. Data are pooled from 2 independent experiments (mean ± SEM). (H) OCR values after etomoxir treatment were subtracted from OCR values before etomoxir treatment in WT and miR-142–/– DCs treated with or without LPS. Data were pooled from 2 independent experiments (mean ± SEM). Comparisons between 2 groups were calculated using paired Student’s t test (2 tailed), while comparisons between 2 groups at multiple time points were calculated utilizing multiple t tests (Holm-Šidák method). *P < 0.05; **P < 0.01.