The genetic deletion of the Dual Specificity Phosphatase 3 (DUSP3) attenuates kidney damage and inflammation following ischaemia/reperfusion injury in mouse

Badr Khbouz; Pascal Rowart; Laurence Poma; Eileen Dahlke; Martina Bottner; Matthew Stokes; Géraldine Bolen; Souad Rahmouni; Franziska Theilig; François Jouret

doi:10.1111/apha.13735

The genetic deletion of the Dual Specificity Phosphatase 3 (DUSP3) attenuates kidney damage and inflammation following ischaemia/reperfusion injury in mouse

Acta Physiol (Oxf). 2022 Feb;234(2):e13735. doi: 10.1111/apha.13735. Epub 2021 Nov 9.

Authors

Badr Khbouz¹, Pascal Rowart^{1

2}, Laurence Poma¹, Eileen Dahlke³, Martina Bottner³, Matthew Stokes⁴, Géraldine Bolen⁵, Souad Rahmouni⁶, Franziska Theilig^{3

7}, François Jouret^{1

8}

Affiliations

¹ Groupe Interdisciplinaire de Génoprotéomique Appliquée (GIGA), Cardiovascular Sciences, University of Liège (ULiège), Liège, Belgium.
² Department of Pharmacology and Chemical Biology, School of Medicine, University of Pittsburgh, Pittsburgh, Pennsylvania, USA.
³ Institute of Anatomy, Christian Albrechts-University, Kiel, Germany.
⁴ Cell Signaling Technology, Inc., Danvers, Massachusetts, USA.
⁵ Department of Clinical Sciences, Fundamental and Applied Research for Animals & Health (FARAH), Veterinary Faculty, University of Liège (ULiège), Liège, Belgium.
⁶ Groupe Interdisciplinaire de Génoprotéomique Appliquée (GIGA), Medical Genomics, University of Liège (ULiège), Liège, Belgium.
⁷ Institute of Anatomy, Department of Medicine, University of Fribourg, Fribourg, Switzerland.
⁸ Division of Nephrology, CHU of Liège, University of Liège (CHU ULiège), Liège, Belgium.

PMID: 34704357
DOI: 10.1111/apha.13735

Abstract

Aim: Dual Specificity Phosphatase 3 (DUSP3) regulates the innate immune response, with a putative role in angiogenesis. Modulating inflammation and perfusion contributes to renal conditioning against ischaemia/reperfusion (I/R). We postulate that the functional loss of DUSP3 is associated with kidney resistance to I/R.

Methods: Ten C57BL/6 male WT and Dusp3^-/- mice underwent right nephrectomy and left renal I/R (30 min/48 hours). Renal injury was assessed based on serum levels of urea (BUN) and Jablonski score. The expression of CD31 and VEGF vascular markers was quantified by RT-qPCR and immuno-staining. Renal resistivity index (RRI) was measured in vivo by Doppler ultrasound. Comparative phosphoproteomics was conducted using IMAC enrichment of phosphopeptides. Inflammatory markers were quantified at both mRNA and protein levels in ischaemic vs non-ischaemic kidneys in WT vs Dusp3^-/- .

Results: At baseline, we located DUSP3 in renal glomeruli and endothelial cells. CD31-positive vascular network was significantly larger in Dusp3^-/- kidneys compared to WT, with a lower RRI in Dusp3^-/- mice. Following I/R, BUN and Jablonski score were significantly lower in Dusp3^-/- vs WT mice. Phosphoproteomics highlighted a down-regulation of inflammatory pathways and up-regulation of phospho-sites involved in cell metabolism and VEGF-related angiogenesis in Dusp3^-/- vs WT ischaemic kidneys. Dusp3^-/- ischaemic kidneys showed decreased mRNA levels of CD11b, TNF-α, KIM-1, IL-6, IL-1β and caspase-3 compared to controls. The numbers of PCNA-, F4-80- and CD11b-positive cells were reduced in Dusp3^-/- vs WT kidneys post-I/R.

Conclusion: Genetic inactivation of Dusp3 is associated with kidney conditioning against I/R, possibly due to attenuated inflammation and improved perfusion.

Keywords: DUSP3; acute kidney injury; kidney; phosphoproteomics; renal ischaemia/reperfusion; sterile inflammation.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Acute Kidney Injury* / metabolism
Animals
Dual Specificity Phosphatase 3* / genetics
Endothelial Cells / metabolism
Inflammation / genetics
Inflammation / metabolism
Kidney / metabolism
Male
Mice
Mice, Inbred C57BL
Reperfusion Injury* / genetics
Reperfusion Injury* / metabolism

Substances

Dual Specificity Phosphatase 3
Dusp3 protein, mouse