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Sci Rep. 2019 Mar 18;9(1):4707. doi: 10.1038/s41598-019-41247-5.

A critical regulator of Bcl2 revealed by systematic transcript discovery of lncRNAs associated with T-cell differentiation.

Author information

1
Aix-Marseille University, Inserm, TAGC, UMR1090, Marseille, France.
2
Equipe Labélisée Ligue Contre le Cancer, Marseille, France.
3
Laboratory of Applied Molecular Biology and Immunology, W0414100, University of Tlemcen, Tlemcen, Algeria.
4
Vinmec Research Institute of Stem cell and Gene technology (VRISG), Hanoi, Vietnam.
5
Molecular Biology and Genetics Laboratory, Dubai, United Arab Emirates.
6
Aix-Marseille University, Inserm, TAGC, UMR1090, Marseille, France. denis.puthier@univ-amu.fr.
7
Equipe Labélisée Ligue Contre le Cancer, Marseille, France. denis.puthier@univ-amu.fr.
8
Aix-Marseille University, Inserm, TAGC, UMR1090, Marseille, France. salvatore.spicuglia@inserm.fr.
9
Equipe Labélisée Ligue Contre le Cancer, Marseille, France. salvatore.spicuglia@inserm.fr.

Abstract

Normal T-cell differentiation requires a complex regulatory network which supports a series of maturation steps, including lineage commitment, T-cell receptor (TCR) gene rearrangement, and thymic positive and negative selection. However, the underlying molecular mechanisms are difficult to assess due to limited T-cell models. Here we explore the use of the pro-T-cell line P5424 to study early T-cell differentiation. Stimulation of P5424 cells by the calcium ionophore ionomycin together with PMA resulted in gene regulation of T-cell differentiation and activation markers, partially mimicking the CD4-CD8- double negative (DN) to double positive (DP) transition and some aspects of subsequent T-cell maturation and activation. Global analysis of gene expression, along with kinetic experiments, revealed a significant association between the dynamic expression of coding genes and neighbor lncRNAs including many newly-discovered transcripts, thus suggesting potential co-regulation. CRISPR/Cas9-mediated genetic deletion of Robnr, an inducible lncRNA located downstream of the anti-apoptotic gene Bcl2, demonstrated a critical role of the Robnr locus in the induction of Bcl2. Thus, the pro-T-cell line P5424 is a powerful model system to characterize regulatory networks involved in early T-cell differentiation and maturation.

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