miR-17∼92 family clusters control iNKT cell ontogenesis via modulation of TGF-β signaling
Edited by Wayne M. Yokoyama, Washington University School of Medicine, St. Louis, MO, and approved November 15, 2016 (received for review July 21, 2016)
Significance
CD1d-restricted invariant natural killer T (iNKT) cells are innate-like T lymphocytes that play fundamental roles in cancer, autoimmunity, and infections. iNKT cells acquire effector functions already in the thymus, because of a distinct developmentally regulated genetic program that is critically controlled by miRNAs. Our study unveils the unexpected requirement for miRNA-dependent fine-tuning of TGF-β signaling in the control of iNKT cell development and functional differentiation. The targeting of a lineage-specific cytokine signaling by miRNA represents a previously unknown level of developmental regulation in the thymus. Furthermore, our study provides a comprehensive atlas of miRNA-regulated molecular pathways involved in iNKT cell ontogenesis, and highlights molecular pathways targeted by defined miRNAs that are predicted to be involved in the development and maturation of CD1d-restricted iNKT cells.
Abstract
Invariant natural killer T cells (iNKT) cells are T lymphocytes displaying innate effector functions, acquired through a distinct thymic developmental program regulated by microRNAs (miRNAs). Deleting miRNAs by Dicer ablation (Dicer KO) in thymocytes selectively impairs iNKT cell survival and functional differentiation. To unravel this miRNA-dependent program, we systemically identified transcripts that were differentially expressed between WT and Dicer KO iNKT cells at different differentiation stages and predicted to be targeted by the iNKT cell-specific miRNAs. TGF-β receptor II (TGF-βRII), critically implicated in iNKT cell differentiation, was found up-regulated in iNKT Dicer KO cells together with enhanced TGF-β signaling. miRNA members of the miR-17∼92 family clusters were predicted to target Tgfbr2 mRNA upon iNKT cell development. iNKT cells lacking all three miR-17∼92 family clusters (miR-17∼92, miR-106a∼363, miR-106b∼25) phenocopied both increased TGF-βRII expression and signaling, and defective effector differentiation, displayed by iNKT Dicer KO cells. Consistently, genetic ablation of TGF-β signaling in the absence of miRNAs rescued iNKT cell differentiation. These results elucidate the global impact of miRNAs on the iNKT cell developmental program and uncover the targeting of a lineage-specific cytokine signaling by miRNAs as a mechanism regulating innate-like T-cell development and effector differentiation.
Data Availability
Data deposition: The data reported in this paper have been deposited in the Gene Expression Omnibus (GEO) database, www.ncbi.nlm.nih.gov/geo (accession no. GSE79901).
Acknowledgments
We thank Giovanni Tonon for the critical reading of the manuscript, and the NIH Tetramer Core Facility for mouse CD1d tetramer. The study was funded by Fondazione Cariplo Grant 2009–3603 (to S.A. and P.D.) and Italian Association for Cancer Research Grant AIRC IG15466 and IG15517 (to P.D. and G.C.).
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Data Availability
Data deposition: The data reported in this paper have been deposited in the Gene Expression Omnibus (GEO) database, www.ncbi.nlm.nih.gov/geo (accession no. GSE79901).
Submission history
Published online: December 5, 2016
Published in issue: December 20, 2016
Keywords
Acknowledgments
We thank Giovanni Tonon for the critical reading of the manuscript, and the NIH Tetramer Core Facility for mouse CD1d tetramer. The study was funded by Fondazione Cariplo Grant 2009–3603 (to S.A. and P.D.) and Italian Association for Cancer Research Grant AIRC IG15466 and IG15517 (to P.D. and G.C.).
Notes
This article is a PNAS Direct Submission.
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The authors declare no conflict of interest.
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miR-17∼92 family clusters control iNKT cell ontogenesis via modulation of TGF-β signaling, Proc. Natl. Acad. Sci. U.S.A.
113 (51) E8286-E8295,
https://doi.org/10.1073/pnas.1612024114
(2016).
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