Mature T cell responses are controlled by microRNA-142
Yaping Sun, … , Thomas Saunders, Pavan Reddy
Yaping Sun, … , Thomas Saunders, Pavan Reddy
Published June 22, 2015
Citation Information: J Clin Invest. 2015;125(7):2825-2840. https://doi.org/10.1172/JCI78753.
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Mature T cell responses are controlled by microRNA-142

Abstract

T cell proliferation is critical for immune responses; however, the molecular mechanisms that mediate the proliferative response are poorly understood. MicroRNAs (miRs) regulate various molecular processes, including development and function of the immune system. Here, utilizing multiple complementary genetic and molecular approaches, we investigated the contribution of a hematopoietic-specific miR, miR-142, in regulating T cell responses. T cell development was not affected in animals with a targeted deletion of Mir142; however, T cell proliferation was markedly reduced following stimulation both in vitro and in multiple murine models of graft-versus-host disease (GVHD). miR-142–deficient T cells demonstrated substantial cell-cycling defects, and microarray and bioinformatics analyses revealed upregulation of genes involved in cell cycling. Moreover, 2 predicted miR-142 target genes, the atypical E2F transcription factors E2f7 and E2f8, were most highly upregulated in miR-142–deficient cells. Clustered regularly interspaced short palindromic repeat interference–mediated (CRISPRi-mediated) silencing of E2F7 and E2F8 in miR-142–deficient T cells ameliorated cell-cycling defects and reduced GVHD, and overexpression of these factors in WT T cells inhibited the proliferative response. Together, these results identify a link between hematopoietic-specific miR-142 and atypical E2F transcription factors in the regulation of mature T cell cycling and suggest that targeting this interaction may be relevant for mitigating GVHD.

Authors

Yaping Sun, Katherine Oravecz-Wilson, Nathan Mathewson, Ying Wang, Richard McEachin, Chen Liu, Tomomi Toubai, Julia Wu, Corinne Rossi, Thomas Braun, Thomas Saunders, Pavan Reddy

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Figure 7

miR-142 regulates T cell cycling in E2F7/E2F8-dependent manner.

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miR-142 regulates T cell cycling in E2F7/E2F8-dependent manner. (A) Impa...
(A) Impact of overexpression of atypical E2F proteins on cell cycle in WT T cells. WT T cells were infected with E2F7/E2F8 lentiviral particles, treated with anti-CD3 and anti-CD28 Abs, and analyzed for cell cycle. Combined data were from 3 independent experiments. (B) Overexpression of E2F7/E2F8 significantly inhibited T cell proliferation, as demonstrated by 3H-TdR incorporation. Data represent combination of 3 independent experiments. (C) Impact of E2F7/E2F8 silencing on cell cycle in miR-142 KO T cells. miR-142 KO T cells were targeted for E2F7/E2F8 silencing as described in Methods, then treated with anti-CD3 and anti-CD28 Abs for 0 to 3 days. Cell-cycle analyses were processed. Combined data were from 3 independent experiments. (D) KD of E2F7/E2F8 in miR-142 KO T cells significantly improved cell proliferation, as demonstrated by 3H-TdR incorporation. Data represent combination of 3 independent experiments. (E) Essential role of E2F7/E2F8 in cell-cycle activities in WT T cells. WT T cells were targeted for E2F7/E2F8 silencing, treated with anti-CD3 and anti-CD28 Abs for 0 to 4 days, and processed for cell-cycle analyses as in C. Data were combined from 3 independent experiments. (F) KD of E2F7/E2F8 in WT T cells significantly enhanced cell proliferation as demonstrated by 3H-TdR incorporation. Data represent combination of 3 independent experiments. Data are shown as mean ± SEM. *P < 0.05; **P < 0.01, Holm-Sidak method (A, C, and E); unpaired t test (B, D, and F).