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Role and therapeutic potential of dietary ketone bodies in lymph vessel growth

Abstract

Lymphatic vessels (LVs), lined by lymphatic endothelial cells (LECs), are indispensable for life1. However, the role of metabolism in LECs has been incompletely elucidated. In the present study, it is reported that LEC-specific loss of OXCT1, a key enzyme of ketone body oxidation2, reduces LEC proliferation, migration and vessel sprouting in vitro and impairs lymphangiogenesis in development and disease in Prox1ΔOXCT1 mice. Mechanistically, OXCT1 silencing lowers acetyl-CoA levels, tricarboxylic acid cycle metabolite pools, and nucleotide precursor and deoxynucleotide triphosphate levels required for LEC proliferation. Ketone body supplementation to LECs induces the opposite effects. Notably, elevation of lymph ketone body levels by a high-fat, low-carbohydrate ketogenic diet or by administration of the ketone body β-hydroxybutyrate increases lymphangiogenesis after corneal injury and myocardial infarction. Intriguingly, in a mouse model of microsurgical ablation of LVs in the tail, which repeats features of acquired lymphoedema in humans, the ketogenic diet improves LV function and growth, reduces infiltration of anti-lymphangiogenic immune cells and decreases oedema, suggesting a novel dietary therapeutic opportunity.

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Fig. 1: KBO regulates lymphangiogenesis.
Fig. 2: KBO promotes lymphangiogenesis in vivo.
Fig. 3: KBO contributes to sustaining the TCA cycle and to dNTP synthesis.
Fig. 4: Effect of HFLC-KD supplementation on mouse tail lymphoedema.

Data availability

All data generated or analysed during this study are included in this published article (and its Supplementary Information). The raw data that support the findings of this study are available from the corresponding author on reasonable request. Supplementary Figs. 1, 5 and 6 have associated raw data (uncropped blots) in Supplementary Fig. 7. Figure 3 and Supplementary Fig. 4 have associated raw data (Excel files) with metabolite abundances.

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Acknowledgements

We thank P. Crawford and T. Mäkinen for providing Oxct1lox/lox and Prox1-creERT2 mice, respectively. We thank G. Bogaert for providing human foreskins. We also thank S.M. Fendt for discussion and advice. This work was supported by fellowships from LE&RN/FDRS (A.Z.), and supporting grants from IUAP P7/03 (P.C.), Methusalem funding by the Flemish government (P.C.), FWO (G.0598.12, G.0532.10, G.0817.11, G.0834.13, to P.C.), Leducq Transatlantic Network Artemis (P.C.), AXA Research Fund (no. 1465, to P.C.), Foundation against Cancer (P.C.), Fund for Translation Biomedical Research (to P.C.), ERC Advanced Research Grant (EU-ERC269073, to P.C.). We thank A. Van Nuffelen, A. Carton, A. Manderveld, K. Brepoels, K. Peeters, N. Dai, M. Rifaad, M. Parys, I. Betz, C. De Legher, S. Wyns, P.J. Coolen, M. Nijs, P. Vanwesemael, B. Verherstraeten, G. Dubois, E. Van Dyck, A. Acosta Sanchez and D. Verdegem for their technical assistance, and various laboratory members for their feedback and discussions.

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M.G.C., A.Z., J.S., A.-C.K.T., L.-A.T., W.V., A.B., R.M.P., S.V., I.C. and B.G. performed research and/or analysed the data. P.d.Z. managed the collaboration with UZ Leuven, wrote the documentation for the Medical Ethical Committee agreement and the informed consent for HDLEC subjects. M.G.C., A.Z., R.M.P., M.M., G.E., M.D. and P.C. designed experiments. M.G.C., A.Z. and P.C. wrote the paper. P.C. conceptualized and supervised the study. All authors discussed the results and commented on the manuscript.

Corresponding author

Correspondence to Peter Carmeliet.

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The authors declare no competing financial or non-financial interests in relation to the work described.

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Supplementary information

Supplementary Information

Supplementary Figs. 1–7

Reporting Summary

Supplementary Table 1

Metabolite abundances in control and OXCT1KD HDLECs measured by MS. Raw metabolite abundances are expressed as arbitrary units normalized to micrograms of protein content

Supplementary Table 2

Metabolite abundances in control and OXCT1KD2 HDLECs measured by MS. Raw metabolite abundances are expressed as arbitrary units normalized to micrograms of protein content

Supplementary Table 3

Metabolite abundances in control and supplemented HDLECs measured by MS. Raw metabolite abundances are expressed as arbitrary units normalized to micrograms of protein content

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García-Caballero, M., Zecchin, A., Souffreau, J. et al. Role and therapeutic potential of dietary ketone bodies in lymph vessel growth. Nat Metab 1, 666–675 (2019). https://doi.org/10.1038/s42255-019-0087-y

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