|
Status |
Public on Dec 31, 2021 |
Title |
The volume regulated anion channel LRRC8C suppresses T cell function by regulating cyclic dinucleotide transport and STING-p53 signaling |
Organism |
Mus musculus |
Experiment type |
Expression profiling by high throughput sequencing
|
Summary |
The volume-regulated anion channel (VRAC) is a hexameric complex formed by LRRC8 proteins. VRACs are responsible for the regulatory volume decrease (RVD) after hypotonic cell swelling by mediating the efflux of chloride. Besides chloride, LRRC8 proteins transport other molecules including immunomodulatory cyclic dinucleotides (CDNs) such as 2’3’cGAMP. Here we identify LRRC8C as a critical component of VRACs in T cells, where its deletion abolishes VRAC currents and RVD. We find that LRRC8C mediates the transport of 2’3’cGAMP in T cells. T cells of Lrrc8c-/- mice have increased cell cycle progression, proliferation, survival, Ca2+ influx and cytokine production in vitro and enhanced T cell mediated immunity in vivo. We used RNA sequencing of CD4+ T cells from wild-type (WT) and Lrrc8c-/- mice to determine the effects of impaired cGAMP signaling in T cells. T cells of Lrrc8c-/- mice had impaired p53 signaling which was associated with decreased p53 expression. We found that uptake of 2’3’cGAMP and other CDNs via LRRC8C results in STING activation and p53 accumulation. Inhibition of STING in WT T cells recapitulates the phenotype of LRRC8C-deficient T cells whereas overexpression of p53 inhibits enhanced T cell function in the absence of LRRC8C. These findings establish cGAMP uptake through LRRC8C and subsequent STING-p53 signaling as a novel inhibitory signaling pathway in T cells and adaptive immunity.
|
|
|
Overall design |
Purified CD4+ T cells from WT and Lrrc8c-/- mice were isolated from the spleen and left unstimulated or stimulated with plate-bound anti-CD3 and anti-CD28 antibodies in 12-well plates for 24h and 48h. Additionally, WT CD4+ T cells were stimulated in the presence of 20 micromolar DCPIB (VRAC inhibitor). A total of 24 samples were analyzed by RNASeq.
|
|
|
Contributor(s) |
Concepcion AR, Khodadadi-Jamayran A, Feske S |
Citation(s) |
35105987 |
|
Submission date |
Dec 21, 2020 |
Last update date |
Feb 05, 2022 |
Contact name |
Alireza Khodadadi-Jamayran |
Organization name |
New York University, NYU Langone Medical Center
|
Department |
Division of Advanced Research Technologies (DART)
|
Lab |
Applied Bioinformatics Laboratories (ABL)
|
Street address |
550 1st Ave, MSB 304
|
City |
New York City |
State/province |
NY |
ZIP/Postal code |
10016 |
Country |
USA |
|
|
Platforms (1) |
GPL17021 |
Illumina HiSeq 2500 (Mus musculus) |
|
Samples (24)
|
|
Relations |
BioProject |
PRJNA687046 |
SRA |
SRP298798 |