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Genetic variations in A20 DUB domain provide a genetic link to citrullination and neutrophil extracellular traps in systemic lupus erythematosus

Odqvist, Lina ; Jevnikar, Zala ; Riise, Rebecca ; Öberg, Lisa ; Rhedin, Magdalena ; Leonard, Dag ; Yrlid, Linda ; Jackson, Sonya ; Mattsson, Johan and Nanda, Sambit , et al. (2019) In Annals of the Rheumatic Diseases 78(10). p.1363-1370
Abstract

Objectives: Genetic variations in TNFAIP3 (A20) de-ubiquitinase (DUB) domain increase the risk of systemic lupus erythematosus (SLE) and rheumatoid arthritis. A20 is a negative regulator of NF-κB but the role of its DUB domain and related genetic variants remain unclear. We aimed to study the functional effects of A20 DUB-domain alterations in immune cells and understand its link to SLE pathogenesis. Methods: CRISPR/Cas9 was used to generate human U937 monocytes with A20 DUB-inactivating C103A knock-in (KI) mutation. Whole genome RNA-sequencing was used to identify differentially expressed genes between WT and C103A KI cells. Functional studies were performed in A20 C103A U937 cells and in immune cells from A20 C103A mice and genotyped... (More)

Objectives: Genetic variations in TNFAIP3 (A20) de-ubiquitinase (DUB) domain increase the risk of systemic lupus erythematosus (SLE) and rheumatoid arthritis. A20 is a negative regulator of NF-κB but the role of its DUB domain and related genetic variants remain unclear. We aimed to study the functional effects of A20 DUB-domain alterations in immune cells and understand its link to SLE pathogenesis. Methods: CRISPR/Cas9 was used to generate human U937 monocytes with A20 DUB-inactivating C103A knock-in (KI) mutation. Whole genome RNA-sequencing was used to identify differentially expressed genes between WT and C103A KI cells. Functional studies were performed in A20 C103A U937 cells and in immune cells from A20 C103A mice and genotyped healthy individuals with A20 DUB polymorphism rs2230926. Neutrophil extracellular trap (NET) formation was addressed ex vivo in neutrophils from A20 C103A mice and SLE-patients with rs2230926. Results: Genetic disruption of A20 DUB domain in human and murine myeloid cells did not give rise to enhanced NF-κB signalling. Instead, cells with C103A mutation or rs2230926 polymorphism presented an upregulated expression of PADI4, an enzyme regulating protein citrullination and NET formation, two key mechanisms in autoimmune pathology. A20 C103A cells exhibited enhanced protein citrullination and extracellular trap formation, which could be suppressed by selective PAD4 inhibition. Moreover, SLE-patients with rs2230926 showed increased NETs and increased frequency of autoantibodies to citrullinated epitopes. Conclusions: We propose that genetic alterations disrupting the A20 DUB domain mediate increased susceptibility to SLE through the upregulation of PADI4 with resultant protein citrullination and extracellular trap formation.

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organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
NET, PAD4, PADI4, peptidyl arginine deiminase, rs2230926
in
Annals of the Rheumatic Diseases
volume
78
issue
10
pages
1363 - 1370
publisher
BMJ Publishing Group
external identifiers
  • scopus:85069535132
  • pmid:31300459
ISSN
0003-4967
DOI
10.1136/annrheumdis-2019-215434
language
English
LU publication?
yes
id
825a9abe-c61e-4ba0-a3ff-544659cbe364
date added to LUP
2019-08-12 10:07:12
date last changed
2021-03-03 05:18:02
@article{825a9abe-c61e-4ba0-a3ff-544659cbe364,
  abstract     = {<p>Objectives: Genetic variations in TNFAIP3 (A20) de-ubiquitinase (DUB) domain increase the risk of systemic lupus erythematosus (SLE) and rheumatoid arthritis. A20 is a negative regulator of NF-κB but the role of its DUB domain and related genetic variants remain unclear. We aimed to study the functional effects of A20 DUB-domain alterations in immune cells and understand its link to SLE pathogenesis. Methods: CRISPR/Cas9 was used to generate human U937 monocytes with A20 DUB-inactivating C103A knock-in (KI) mutation. Whole genome RNA-sequencing was used to identify differentially expressed genes between WT and C103A KI cells. Functional studies were performed in A20 C103A U937 cells and in immune cells from A20 C103A mice and genotyped healthy individuals with A20 DUB polymorphism rs2230926. Neutrophil extracellular trap (NET) formation was addressed ex vivo in neutrophils from A20 C103A mice and SLE-patients with rs2230926. Results: Genetic disruption of A20 DUB domain in human and murine myeloid cells did not give rise to enhanced NF-κB signalling. Instead, cells with C103A mutation or rs2230926 polymorphism presented an upregulated expression of PADI4, an enzyme regulating protein citrullination and NET formation, two key mechanisms in autoimmune pathology. A20 C103A cells exhibited enhanced protein citrullination and extracellular trap formation, which could be suppressed by selective PAD4 inhibition. Moreover, SLE-patients with rs2230926 showed increased NETs and increased frequency of autoantibodies to citrullinated epitopes. Conclusions: We propose that genetic alterations disrupting the A20 DUB domain mediate increased susceptibility to SLE through the upregulation of PADI4 with resultant protein citrullination and extracellular trap formation.</p>},
  author       = {Odqvist, Lina and Jevnikar, Zala and Riise, Rebecca and Öberg, Lisa and Rhedin, Magdalena and Leonard, Dag and Yrlid, Linda and Jackson, Sonya and Mattsson, Johan and Nanda, Sambit and Cohen, Philip and Knebel, Axel and Arthur, Simon and Thörn, Kristofer and Svenungsson, Elisabet and Jönsen, Andreas and Gunnarsson, Iva and Tandre, Karolina and Alexsson, Andrei and Kastbom, Alf and Rantapää-Dahlqvist, Solbritt and Eloranta, Maija Leena and Syvänen, Ann Christine and Bengtsson, Anders and Johansson, Patrik and Sandling, Johanna K. and Sjöwall, Christopher and Rönnblom, Lars and Collins, Barry and Vaarala, Outi},
  issn         = {0003-4967},
  language     = {eng},
  month        = {07},
  number       = {10},
  pages        = {1363--1370},
  publisher    = {BMJ Publishing Group},
  series       = {Annals of the Rheumatic Diseases},
  title        = {Genetic variations in A20 DUB domain provide a genetic link to citrullination and neutrophil extracellular traps in systemic lupus erythematosus},
  url          = {http://dx.doi.org/10.1136/annrheumdis-2019-215434},
  doi          = {10.1136/annrheumdis-2019-215434},
  volume       = {78},
  year         = {2019},
}