Prolyl-4-hydroxylase domain 3 (PHD3) is a critical terminator for cell survival of macrophages under stress conditions

Lija Swain; Marieke Wottawa; Annette Hillemann; Angelika Beneke; Haruki Odagiri; Kazutoyo Terada; Motoyoshi Endo; Yuichi Oike; Katja Farhat; Dörthe M Katschinski

doi:10.1189/jlb.2HI1013-533R

Prolyl-4-hydroxylase domain 3 (PHD3) is a critical terminator for cell survival of macrophages under stress conditions

J Leukoc Biol. 2014 Sep;96(3):365-75. doi: 10.1189/jlb.2HI1013-533R. Epub 2014 Mar 13.

Affiliations

¹ Institute of Cardiovascular Physiology, University Medical Center, Georg August University Göttingen, Germany; and.
² Department of Molecular Genetics, Graduate School of Medical Sciences, Kumamoto University, Japan.
³ Institute of Cardiovascular Physiology, University Medical Center, Georg August University Göttingen, Germany; and [email protected].

Abstract

On a molecular level, cells sense changes in oxygen availability through the PHDs, which regulate the protein stability of the α-subunit of the transcription factor HIF. Especially, PHD3 has been additionally associated with apoptotic cell death. We hypothesized that PHD3 plays a role in cell-fate decisions in macrophages. Therefore, myeloid-specific PHD3(-/-) mice were created and analyzed. PHD3(-/-) BMDM showed no altered HIF-1α or HIF-2α stabilization or increased HIF target gene expression in normoxia or hypoxia. Macrophage M1 and M2 polarization was unchanged likewise. Compared with macrophages from WT littermates, PHD3(-/-) BMDM exhibited a significant reduction in TUNEL-positive cells after serum withdrawal or treatment with stauro and SNAP. Under the same conditions, PHD3(-/-) BMDM also showed less Annexin V staining, which is representative for membrane disruption, and indicated a reduced early apoptosis. In an unbiased transcriptome screen, we found that Angptl2 expression was reduced in PHD3(-/-) BMDM under stress conditions. Addition of rAngptl2 rescued the antiapoptotic phenotype, demonstrating that it is involved in the PHD3-mediated response toward apoptotic stimuli in macrophages.

Keywords: HIF; angiopoietin-like protein; hypoxia; oxygen sensing.

Publication types

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

MeSH terms

Angiopoietin-Like Protein 2
Angiopoietin-like Proteins
Angiopoietins / biosynthesis
Angiopoietins / genetics
Angiopoietins / pharmacology
Animals
Apoptosis
Basic Helix-Loop-Helix Transcription Factors / physiology
Bone Marrow Cells / cytology
Cell Hypoxia
Cells, Cultured
Gene Expression Regulation
Hydroxylation
Hypoxia-Inducible Factor 1, alpha Subunit / physiology
Macrophages / cytology*
Macrophages / drug effects
Macrophages / enzymology
Mice
Mice, Inbred C57BL
Mice, Knockout
Myeloid Cells / enzymology
NF-kappa B / metabolism
Procollagen-Proline Dioxygenase / deficiency
Procollagen-Proline Dioxygenase / genetics
Procollagen-Proline Dioxygenase / physiology*
Protein Processing, Post-Translational
Recombinant Fusion Proteins / pharmacology
S-Nitroso-N-Acetylpenicillamine / pharmacology
Staurosporine / pharmacology
Transcription, Genetic
Transcriptome

Substances

ANGPTL2 protein, human
Angiopoietin-Like Protein 2
Angiopoietin-like Proteins
Angiopoietins
Angptl2 protein, mouse
Basic Helix-Loop-Helix Transcription Factors
Hif1a protein, mouse
Hypoxia-Inducible Factor 1, alpha Subunit
NF-kappa B
Recombinant Fusion Proteins
endothelial PAS domain-containing protein 1
S-Nitroso-N-Acetylpenicillamine
PHD3 protein, mouse
Procollagen-Proline Dioxygenase
Staurosporine