PUBLICATION

A zebrafish functional genomics model to investigate the role of human A20 variants in vivo

Authors
Cultrone, D., Zammit, N.W., Self, E., Postert, B., Han, J.Z.R., Bailey, J., Warren, J., Croucher, D.R., Kikuchi, K., Bogdanovic, O., Chtanova, T., Hesselson, D., Grey, S.T.
ID
ZDB-PUB-201120-49
Date
2020
Source
Scientific Reports   10: 19085 (Journal)
Registered Authors
Bogdanovic, Ozren, Cultrone, Daniele, Grey, Shane, Hesselson, Daniel, Kikuchi, Kazu, Postert, Benno
Keywords
none
MeSH Terms
  • Macrophages/immunology
  • Macrophages/metabolism
  • Models, Genetic
  • Recombinant Proteins/genetics
  • Recombinant Proteins/metabolism
  • Amino Acid Substitution
  • Tumor Necrosis Factor alpha-Induced Protein 3/deficiency
  • Tumor Necrosis Factor alpha-Induced Protein 3/genetics*
  • Tumor Necrosis Factor alpha-Induced Protein 3/physiology*
  • Phosphorylation
  • Animals
  • NF-kappa B/metabolism
  • Humans
  • Zebrafish Proteins/deficiency
  • Zebrafish Proteins/genetics*
  • Zebrafish Proteins/physiology*
  • Zebrafish/genetics*
  • Zebrafish/physiology
  • Conserved Sequence
  • Evolution, Molecular
  • Genetic Variation
  • Autoimmune Diseases/etiology
  • Autoimmune Diseases/genetics
  • Inflammation/etiology
  • Inflammation/genetics
  • Models, Animal
  • Animals, Genetically Modified
  • Mutation, Missense
(all 28)
PubMed
33154446 Full text @ Sci. Rep.
Abstract
Germline loss-of-function variation in TNFAIP3, encoding A20, has been implicated in a wide variety of autoinflammatory and autoimmune conditions, with acquired somatic missense mutations linked to cancer progression. Furthermore, human sequence data reveals that the A20 locus contains ~ 400 non-synonymous coding variants, which are largely uncharacterised. The growing number of A20 coding variants with unknown function, but potential clinical impact, poses a challenge to traditional mouse-based approaches. Here we report the development of a novel functional genomics approach that utilizes a new A20-deficient zebrafish (Danio rerio) model to investigate the impact of TNFAIP3 genetic variants in vivo. A20-deficient zebrafish are hyper-responsive to microbial immune activation and exhibit spontaneous early lethality. Ectopic addition of human A20 rescued A20-null zebrafish from lethality, while missense mutations at two conserved A20 residues, S381A and C243Y, reversed this protective effect. Ser381 represents a phosphorylation site important for enhancing A20 activity that is abrogated by its mutation to alanine, or by a causal C243Y mutation that triggers human autoimmune disease. These data reveal an evolutionarily conserved role for TNFAIP3 in limiting inflammation in the vertebrate linage and show how this function is controlled by phosphorylation. They also demonstrate how a zebrafish functional genomics pipeline can be utilized to investigate the in vivo significance of medically relevant human TNFAIP3 gene variants.
Genes / Markers
Figures
Figure Gallery (4 images)
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Expression
Phenotype
Fish Conditions Stage Phenotype Figure
WTchemical treatment by environment: lipopolysaccharideDay 6
tnfaip3dgi4/+; gi5Tgstandard conditionsProtruding-mouth
tnfaip3dgi4/+; gi5Tgchemical treatment by environment: lipopolysaccharideDay 6
tnfaip3dgi4/+; vcc7Tgstandard conditionsDays 7-13
tnfaip3dgi4/+; vcc7Tgstandard conditionsDays 7-13
tnfaip3dgi4/dgi4standard conditionsDays 14-20
tnfaip3dgi4/dgi4standard conditionsDays 14-20
tnfaip3dgi4/dgi4; gi5Tgstandard conditionsProtruding-mouth
tnfaip3dgi4/dgi4; gi5Tgchemical treatment by environment: lipopolysaccharideDay 6
tnfaip3dgi4/dgi4; gz15Tg; m1018Tgstandard conditionsDays 14-20
1 - 10 of 13
Show
Mutations / Transgenics
Allele Construct Type Affected Genomic Region
dgi4
    		Small Deletion
    gi5TgTransgenic Insertion
      gz15TgTransgenic Insertion
        m1018TgTransgenic Insertion
          vcc4TgTransgenic Insertion
            vcc7TgTransgenic Insertion
              1 - 6 of 6
              Show
              Human Disease / Model
              No data available
              Sequence Targeting Reagents
              No data available
              Fish
              Fish
              gi5Tg
              gi5Tg; vcc7Tg
              tnfaip3dgi4/dgi4
              tnfaip3dgi4/dgi4; gi5Tg
              tnfaip3dgi4/dgi4; gi5Tg; vcc7Tg
              tnfaip3dgi4/dgi4; gz15Tg
              tnfaip3dgi4/dgi4; gz15Tg; m1018Tg
              tnfaip3dgi4/dgi4; vcc4Tg
              tnfaip3dgi4/dgi4; vcc7Tg
              tnfaip3dgi4/+; gi5Tg
              1 - 10 of 14
              Show
              Antibodies
              No data available
              Orthology
              No data available
              Engineered Foreign Genes
              Marker Marker Type Name
              DsRedEFGDsRed
              EGFPEFGEGFP
              1 - 2 of 2
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              Mapping
              No data available
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              Citation
              Cultrone, D., Zammit, N.W., Self, E., Postert, B., Han, J.Z.R., Bailey, J., Warren, J., Croucher, D.R., Kikuchi, K., Bogdanovic, O., Chtanova, T., Hesselson, D., Grey, S.T. (2020) A zebrafish functional genomics model to investigate the role of human A20 variants in vivo. Scientific Reports. 10:19085.