Loss of zebrafish atp6v1e1b, encoding a subunit of vacuolar ATPase, recapitulates human ARCL type 2C syndrome and identifies multiple pathobiological signatures

Lore Pottie; Wouter Van Gool; Michiel Vanhooydonck; Franz-Georg Hanisch; Geert Goeminne; Andreja Rajkovic; Paul Coucke; Patrick Sips; Bert Callewaert

doi:10.1371/journal.pgen.1009603

Loss of zebrafish atp6v1e1b, encoding a subunit of vacuolar ATPase, recapitulates human ARCL type 2C syndrome and identifies multiple pathobiological signatures

PLoS Genet. 2021 Jun 18;17(6):e1009603. doi: 10.1371/journal.pgen.1009603. eCollection 2021 Jun.

Authors

Lore Pottie^{1

2}, Wouter Van Gool^{1

2}, Michiel Vanhooydonck^{1

2}, Franz-Georg Hanisch³, Geert Goeminne^{4

5}, Andreja Rajkovic⁶, Paul Coucke^{1

2}, Patrick Sips^{1

2}, Bert Callewaert^{1

2}

Affiliations

¹ Center for Medical Genetics Ghent, Ghent University Hospital, Ghent, Belgium.
² Department of Biomolecular Medicine, Ghent University, Ghent, Belgium.
³ Institute of Biochemistry II, Medical Faculty, University of Cologne, Cologne, Germany.
⁴ VIB Metabolomics Core Ghent, Ghent, Belgium.
⁵ Department of Plant Biotechnology and Bioinformatics, Ghent University, Ghent, Belgium.
⁶ Department of Food technology, Safety and Health, Faculty of Bioscience Engineering, University of Ghent, Ghent, Belgium.

Abstract

The inability to maintain a strictly regulated endo(lyso)somal acidic pH through the proton-pumping action of the vacuolar-ATPases (v-ATPases) has been associated with various human diseases including heritable connective tissue disorders. Autosomal recessive (AR) cutis laxa (CL) type 2C syndrome is associated with genetic defects in the ATP6V1E1 gene and is characterized by skin wrinkles or loose redundant skin folds with pleiotropic systemic manifestations. The underlying pathological mechanisms leading to the clinical presentations remain largely unknown. Here, we show that loss of atp6v1e1b in zebrafish leads to early mortality, associated with craniofacial dysmorphisms, vascular anomalies, cardiac dysfunction, N-glycosylation defects, hypotonia, and epidermal structural defects. These features are reminiscent of the phenotypic manifestations in ARCL type 2C patients. Our data demonstrates that loss of atp6v1e1b alters endo(lyso)somal protein levels, and interferes with non-canonical v-ATPase pathways in vivo. In order to gain further insights into the processes affected by loss of atp6v1e1b, we performed an untargeted analysis of the transcriptome, metabolome, and lipidome in early atp6v1e1b-deficient larvae. We report multiple affected pathways including but not limited to oxidative phosphorylation, sphingolipid, fatty acid, and energy metabolism together with profound defects on mitochondrial respiration. Taken together, our results identify complex pathobiological effects due to loss of atp6v1e1b in vivo.

Publication types

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

MeSH terms

Abnormalities, Multiple / genetics*
Abnormalities, Multiple / metabolism
Abnormalities, Multiple / pathology
Animals
Cutis Laxa / genetics*
Cutis Laxa / metabolism
Cutis Laxa / pathology
Disease Models, Animal
Endosomes / metabolism
Endosomes / pathology
Epithelial Cells / metabolism*
Epithelial Cells / pathology
Gene Expression Regulation
Humans
Larva / genetics
Larva / growth & development
Larva / metabolism
Lipidomics
Longevity / genetics
Lysosomes / metabolism
Lysosomes / pathology
Metabolome / genetics
Mitochondria / metabolism
Mitochondria / pathology
Oxidative Phosphorylation
Protein Isoforms / deficiency
Protein Isoforms / genetics
Skin / metabolism*
Skin / pathology
Syndrome
Transcriptome
Vacuolar Proton-Translocating ATPases / deficiency
Vacuolar Proton-Translocating ATPases / genetics*
Zebrafish / embryology
Zebrafish / genetics
Zebrafish / metabolism
Zebrafish Proteins / deficiency
Zebrafish Proteins / genetics*

Substances

Protein Isoforms
Zebrafish Proteins
ATP6V1E2 protein, human
Vacuolar Proton-Translocating ATPases

Grants and funding

B.C. is a senior clinical investigator of the Research Foundation Flanders. This work was supported by a starting grant of the Special Research Fund from Ghent University (grant 01N04516C to B.C.), by a Methusalem Grant (BOFMET2015000401) from Ghent University, by a junior fundamental research project grant of the Fund for Scientific Research (G035620N) and by the European Union’s Horizon 2020 research and innovation program, under the Marie Skłodowska-Curie grant agreement No. 794365 to P.S. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.