PUBLICATION

Genetic and pharmacological inhibition of CDK9 drives neutrophil apoptosis to resolve inflammation in zebrafish in vivo

Authors
Hoodless, L.J., Lucas, C.D., Duffin, R., Denvir, M.A., Haslett, C., Tucker, C.S., Rossi, A.G.
ID
ZDB-PUB-161113-16
Date
2016
Source
Scientific Reports   5: 36980 (Journal)
Registered Authors
Duffin, Rodger, Hoodless, Laura, Lucas, Chris, Rossi, Adriano
Keywords
Acute inflammation, Imaging the immune system, Immune cell death
MeSH Terms
  • Zebrafish
  • Macrophages/cytology
  • Macrophages/drug effects
  • Macrophages/metabolism
  • Pyrazoles/pharmacology
  • Gene Knockdown Techniques/methods*
  • Cyclin-Dependent Kinase 9/genetics*
  • Cyclin-Dependent Kinase 9/metabolism*
  • Flavonoids/pharmacology
  • Disease Models, Animal
  • Ribonucleoproteins/genetics
  • Ribonucleoproteins/metabolism
  • Zebrafish Proteins/genetics*
  • Zebrafish Proteins/metabolism*
  • Protein Kinase Inhibitors/pharmacology*
  • Piperidines/pharmacology
  • Animals
  • Neutrophils/cytology*
  • Neutrophils/drug effects
  • Neutrophils/metabolism
  • Apoptosis/drug effects
  • CRISPR-Cas Systems
  • Inflammation/immunology*
  • Inflammation/metabolism
(all 24)
PubMed
27833165 Full text @ Sci. Rep.
Abstract
Neutrophilic inflammation is tightly regulated and subsequently resolves to limit tissue damage and promote repair. When the timely resolution of inflammation is dysregulated, tissue damage and disease results. One key control mechanism is neutrophil apoptosis, followed by apoptotic cell clearance by phagocytes such as macrophages. Cyclin-dependent kinase (CDK) inhibitor drugs induce neutrophil apoptosis in vitro and promote resolution of inflammation in rodent models. Here we present the first in vivo evidence, using pharmacological and genetic approaches, that CDK9 is involved in the resolution of neutrophil-dependent inflammation. Using live cell imaging in zebrafish with labelled neutrophils and macrophages, we show that pharmacological inhibition, morpholino-mediated knockdown and CRISPR/cas9-mediated knockout of CDK9 enhances inflammation resolution by reducing neutrophil numbers via induction of apoptosis after tailfin injury. Importantly, knockdown of the negative regulator La-related protein 7 (LaRP7) increased neutrophilic inflammation. Our data show that CDK9 is a possible target for controlling resolution of inflammation.
Genes / Markers
Figures
Figure Gallery (7 images)
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Expression
Phenotype
Fish Conditions Stage Phenotype Figure
cdk9ed9/+; i114TgcontrolProtruding-mouth
cdk9ed9/+; i114Tgtransection: caudal finProtruding-mouth
cdk9ed9/+; i114Tgtransection: caudal finDay 4
cdk9ed9/ed9; i114TgcontrolProtruding-mouth
cdk9ed9/ed9; i114Tgtransection: caudal finProtruding-mouth
cdk9ed9/ed9; i114Tgtransection: caudal finDay 4
gl23Tgtransection: caudal fin, chemical treatment by injection: 4-(2,6-dichlorobenzamido)-N-(piperidin-4-yl)-pyrazole-3-carboxamideDay 4
gl23Tg + MO15-cdk9transection: caudal finProtruding-mouth
gl23Tg + MO15-cdk9transection: caudal finDay 4
gl23Tg + MO3-cdk9transection: caudal finProtruding-mouth
1 - 10 of 29
Show
Mutations / Transgenics
Allele Construct Type Affected Genomic Region
ed9
    		Small Deletion
    gl23TgTransgenic Insertion
      i114TgTransgenic Insertion
        1 - 3 of 3
        Show
        Human Disease / Model
        No data available
        Sequence Targeting Reagents
        Fish
        Fish
        cdk9ed9/ed9; i114Tg
        cdk9ed9/+; i114Tg
        gl23Tg
        gl23Tg + MO3-cdk9
        gl23Tg + MO15-cdk9
        i114Tg
        i114Tg + MO2-larp7
        i114Tg + MO3-cdk9
        i114Tg + MO15-cdk9
        1 - 9 of 9
        Show
        Antibodies
        Orthology
        Engineered Foreign Genes
        Marker Marker Type Name
        GFPEFGGFP
        mCherryEFGmCherry
        1 - 2 of 2
        Show
        Mapping
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        Citation
        Hoodless, L.J., Lucas, C.D., Duffin, R., Denvir, M.A., Haslett, C., Tucker, C.S., Rossi, A.G. (2016) Genetic and pharmacological inhibition of CDK9 drives neutrophil apoptosis to resolve inflammation in zebrafish in vivo. Scientific Reports. 5:36980.