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Tumor vessel disintegration by maximum tolerable PFKFB3 blockade

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Abstract

Blockade of the glycolytic activator PFKFB3 in cancer cells (using a maximum tolerable dose of 70 mg/kg of the PFKFB3 blocker 3PO) inhibits tumor growth in preclinical models and is currently being tested as a novel anticancer treatment in phase I clinical trials. However, a detailed preclinical analysis of the effects of such maximum tolerable dose of a PFKFB3 blocker on the tumor vasculature is lacking, even though tumor endothelial cells are hyper-glycolytic. We report here that a high dose of 3PO (70 mg/kg), which inhibits cancer cell proliferation and reduces primary tumor growth, causes tumor vessel disintegration, suppresses endothelial cell growth for protracted periods, (model-dependently) aggravates tumor hypoxia, and compromises vascular barrier integrity, thereby rendering tumor vessels more leaky and facilitating cancer cell intravasation and dissemination. These findings contrast to the effects of a low dose of 3PO (25 mg/kg), which induces tumor vessel normalization, characterized by vascular barrier tightening and maturation, but reduces cancer cell intravasation and metastasis. Our findings highlight the importance of adequately dosing a glycolytic inhibitor for anticancer treatment.

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Acknowledgements

We acknowledge VRC core facilities for their contributions.

Funding

LCC is supported by the Else Kroener-Fresenius-Stiftung and the Fritz Thyssen Stiftung (10.16.2.017MN); AB is supported by an Erasmus Mundus, ERAWEB fellowship (E2.D2.14.289); ARC is supported by the Research Foundation Flanders (FWO 1246617N); AP is supported by an Erwin Schrödinger Fellowship of the Austrian Science Fund FWF (J3730-B26). The work of PC is supported by a Belgian Science Policy Grant (IUAP7/03), long-term structural Methusalem funding by the Flemish Government, Grants from the FWO (G.0834.13N and G.0532.10N), Foundation against Cancer (Grant no. 2012-175), a European Research Council (ERC) Advanced Research Grant (EU ERC269073), and an AXA Research Grant.

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Author notes

  1. Lena-Christin Conradi, Aleksandra Brajic and Anna Rita Cantelmo have contributed equally to this work.

Authors and Affiliations

  1. Laboratory of Angiogenesis and Vascular Metabolism, Department of Oncology, KU Leuven, 3000, Leuven, Belgium

    Lena-Christin Conradi, Aleksandra Brajic, Anna Rita Cantelmo, Ann Bouché, Joanna Kalucka, Andreas Pircher, Ulrike Brüning, Laure-Anne Teuwen, Stefan Vinckier, Mieke Dewerchin & Peter Carmeliet

  2. Laboratory of Angiogenesis and Vascular Metabolism, Vesalius Research Center (VRC), Center for Cancer Biology (CCB), VIB, Campus Gasthuisberg, O&N4; Herestraat 49-B912, 3000, Leuven, Belgium

    Lena-Christin Conradi, Aleksandra Brajic, Anna Rita Cantelmo, Ann Bouché, Joanna Kalucka, Andreas Pircher, Ulrike Brüning, Laure-Anne Teuwen, Stefan Vinckier, Mieke Dewerchin & Peter Carmeliet

  3. Metabolomics Core Facility, Department of Oncology, KU Leuven, 3000, Leuven, Belgium

    Bart Ghesquière

  4. Metabolomics Core Facility, Vesalius Research Center (VRC), Center for Cancer Biology (CCB), VIB, 3000, Leuven, Belgium

    Bart Ghesquière

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  1. Lena-Christin Conradi
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  2. Aleksandra Brajic
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Correspondence to Peter Carmeliet.

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PC declares to be named as inventor on patent applications claiming subject matter related to the findings reviewed in this publication. The other authors declare no conflict of interest.

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All applicable international, national, and/or institutional guidelines for the care and use of animals were followed.

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Figure S1

Concentration-dependent effects of 3PO on EC glycolysis and survival. A, Concentration–response analysis of the effect of 3PO on proliferation of ECs (n = 3). B, Concentration–response analysis of the effect of 3PO on EC cell death measured by LDH release (n = 3). C, Concentration–response analysis of the effect of 3PO on the glycolytic flux of ECs (n = 3). D, E, Morphometric quantification of EC spheroid sprouting, revealing that blockade of PFKFB3 by the high concentration of 3PO (30 μM) reduced total sprout length (D) and number of sprouts per spheroid (E) more pronouncedly than the low concentration of 3PO (10 μM) (mean ± SEM; n = 10). F–H, Representative bright-field micrographs of control (F) and low-concentration (G, 10 μM) and high-concentration (H, 30 μM) 3PO-treated EC spheroids, showing that vascular sprouting is most impaired by the high concentration of 3PO. Scale bars: 50 μm. All quantitative data are mean ± SEM. * p < 0.05. (PDF 143 kb)

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Conradi, LC., Brajic, A., Cantelmo, A.R. et al. Tumor vessel disintegration by maximum tolerable PFKFB3 blockade. Angiogenesis 20, 599–613 (2017). https://doi.org/10.1007/s10456-017-9573-6

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