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Mitochondrial complex III is necessary for endothelial cell proliferation during angiogenesis

Nature Metabolism volume 1, pages 158–171 (2019)Cite this article

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Abstract

Endothelial cells (ECs) require glycolysis for proliferation and migration during angiogenesis; however, the necessity for the mitochondrial respiratory chain during angiogenesis is not known. Here we report that inhibition of respiratory chain complex III impairs proliferation, but not migration, of ECs in vitro by decreasing the NAD+/NADH ratio. To determine whether mitochondrial respiration is necessary for angiogenesis in vivo, we conditionally ablate a subunit of the respiratory chain complex III (QPC) in ECs. Loss of QPC decreases respiration, resulting in diminished EC proliferation, and impairment in retinal and tumour angiogenesis. Loss of QPC does not decrease genes associated with anabolism or nucleotide levels in ECs but diminishes amino acid levels. Our findings indicate that mitochondrial respiration is necessary for angiogenesis and that the primary role of mitochondria in ECs is to serve as biosynthetic organelles for cell proliferation.

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Fig. 1: Mitochondrial complex III is required for endothelial cell proliferation in vitro.
Fig. 2: Mitochondrial complex III maintains NAD+/NADH ratio, necessary for endothelial cell proliferation.
Fig. 3: Mitochondrial complex III respiration in ECs is required for postnatal retinal angiogenesis.
Fig. 4: Mitochondrial complex III respiration in ECs is necessary for developmental angiogenesis.
Fig. 5: Loss of mitochondrial complex III function in ECs increases anabolic-associated gene expression.
Fig. 6: Mitochondrial complex III in ECs is necessary to maintain amino acid levels in vivo.
Fig. 7: Mitochondrial complex III respiration in ECs is required for tumour angiogenesis.

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Data availability

All data from this manuscript are available from the corresponding author upon request. RNA sequence data that support the findings of this study have been deposited in GEO with the accession code GSE121770.

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Acknowledgements

This work was supported by the National Institutes of Health (NIH) grant nos R35CA197532, 5P01AG049665 and 5P01HL071643-13 (to N.S.C.) and T32 GM08061 (to L.P.D.). We thank R. Adams for Cdh5CreERT2 mice. Imaging work was performed at the Northwestern University Center for Advanced Microscopy generously supported by NCI CCSG P30 CA060553 awarded to the Robert H Lurie Comprehensive Cancer Center. Histology services for tumour tissue was provided by the Northwestern University Research Histology and Phenotyping Laboratory, which is supported by NCI P30 CA060553 awarded to the Robert H. Lurie Comprehensive Cancer Center. Analysis of histone modifications was performed by the Northwestern Proteomics Core Facility, generously supported by NCI CCSG P30 CA060553 awarded to the Robert H. Lurie Comprehensive Cancer Center and the National Resource for Translational and Developmental Proteomics supported by P41 GM108569. Flow cytometric analysis was supported by the Northwestern University Flow Cytometry Core Facility supported by NCI CCSG P30 CA060553. Flow cytometry cell sorting was performed on a BD FACSAria SORP system, purchased through the support of NIH grant 1S10OD011996-01. Metabolomics services were performed by the Metabolomics Core Facility at Robert H. Lurie Comprehensive Cancer Center of Northwestern University. We would like to thank H. Abdala-Valencia and K. Nam for RNA sequencing. Finally, we would like to thank G. Oliver at Northwestern University for his helpful intellectual input.

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Authors and Affiliations

  1. Department of Medicine and Robert H. Lurie Cancer Center, Northwestern University Feinberg School of Medicine, Chicago, IL, USA

    Lauren P. Diebold, Samuel E. Weinberg & Navdeep S. Chandel

  2. Center for Vascular and Developmental Biology, Feinberg Cardiovascular Research Institute, Northwestern University, Chicago, IL, USA

    Hyea Jin Gil

  3. Robert H. Lurie Cancer Center Metabolomics Core, Northwestern University Feinberg School of Medicine, Chicago, IL, USA

    Peng Gao

  4. Department of Biochemistry and Molecular Genetics, Northwestern University Feinberg School of Medicine, Chicago, IL, USA

    Carlos A. Martinez

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Contributions

L.P.D. carried out the majority of experiments and data analysis. P.G. performed the HPLCMS/MS metabolite analysis. C.A.M. analysed the data. H.J.G. and S.E.W. provided intellectual and technical expertise. L.P.D. and N.S.C. provided intellectual input and wrote the paper.

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Correspondence to Navdeep S. Chandel.

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Diebold, L.P., Gil, H.J., Gao, P. et al. Mitochondrial complex III is necessary for endothelial cell proliferation during angiogenesis. Nat Metab 1, 158–171 (2019). https://doi.org/10.1038/s42255-018-0011-x

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