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Pleiotrophin selectively binds to vascular endothelial growth factor receptor 2 and inhibits or stimulates cell migration depending on ανβ3 integrin expression

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Abstract

Pleiotrophin (PTN) has a moderate stimulatory effect on endothelial cell migration through ανβ3 integrin, while it decreases the stimulatory effect of vascular endothelial growth factor A (VEGFA) and inhibits cell migration in the absence of ανβ3 through unknown mechanism(s). In the present work, by using a multitude of experimental approaches, we show that PTN binds to VEGF receptor type 2 (VEGFR2) with a KD of 11.6 nM. Molecular dynamics approach suggests that PTN binds to the same VEGFR2 region with VEGFA through its N-terminal domain. PTN inhibits phosphorylation of VEGFR2 at Tyr1175 and still stimulates endothelial cell migration in the presence of a selective VEGFR2 tyrosine kinase inhibitor. VEGFR2 downregulation by siRNA or an anti-VEGFR2 antibody that binds to the ligand-binding VEGFR2 domain also induce endothelial cell migration, which is abolished by a function-blocking antibody against ανβ3 or the peptide PTN112−136 that binds ανβ3 and inhibits PTN binding. In cells that do not express ανβ3, PTN decreases both VEGFR2 Tyr1175 phosphorylation and cell migration in a VEGFR2-dependent manner. Collectively, our data identify VEGFR2 as a novel PTN receptor involved in the regulation of cell migration by PTN and contribute to the elucidation of the mechanism of activation of endothelial cell migration through the interplay between VEGFR2 and ανβ3.

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Acknowledgements

This work has been co-financed by the Operational Program “Human Resources Development, Education and Lifelong Learning” and is co-financed by the European Union (European Social Fund) and Greek national funds, by a Marie Curie Intra European Fellowship within the 7th European Community Framework Programme (ALTangioTARGET, Grant Agreement No. 626057), Andreas Mentzelopoulos family grants scholarship for postgraduate studies at the University of Patras (to PK) and the State Scholarship Foundation in Greece (IKY) (Operational Program “Human Resources Development—Education and Lifelong Learning”, Partnership Agreement (PA) 2014–2020, scholarship to EC). We thank the Advanced Light Microscopy facility of the School of Health Sciences, University of Patras, for using the Leica SP5 confocal microscope. Synthesis of PTN16−24 and PTN112−136 was performed under the supervision of our beloved Prof. Paul Cordopatis, who is deceased.

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Author notes

  1. Marina Koutsioumpa

    Present address: Center for Systems Biomedicine, Vatche and Tamar Manoukian Division of Digestive Diseases, David Geffen School of Medicine, University of California at Los Angeles, Los Angeles, CA, 90095, USA

  2. Evangelia Poimenidi

    Present address: Dual Training in Intensive Care Medicine and Anaesthesia, Glenfield General Hospital, University Hospitals of Leicester NHS Trust, Groby Road, Leicester, LE3 9QP, UK

Authors and Affiliations

  1. Laboratory of Molecular Pharmacology, Department of Pharmacy, University of Patras, 26504, Patras, Greece

    Margarita Lamprou, Pinelopi Kastana, Fani Kofina, Spyridoula Barmpoutsi, Marina Koutsioumpa, Evangelia Poimenidi, Dimitrios Tassopoulos, Effrosyni Choleva & Evangelia Papadimitriou

  2. Department of Chemistry, University of Patras, Patras, Greece

    Ηaralampos Tzoupis & Theodore Tselios

  3. Department of Pharmaceutical Sciences, School of Pharmacy, Texas Tech University Health Sciences Center, Amarillo, TX, USA

    Md Sanaullah Sajib & Constantinos M. Mikelis

  4. Laboratory of Pharmacognosy and Chemistry of Natural Products, Department of Pharmacy, University of Patras, 26504, Patras, Greece

    Aikaterini A. Zompra

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Contributions

ML, PK, FK, ΗT, SB, MSS, MK, EP, DT and AAZ performed experiments, acquired data and tools and analyzed data; ML, TT and CMM analyzed data and critically reviewed the manuscript; EP designed the study, analyzed data and wrote the manuscript.

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Correspondence to Evangelia Papadimitriou.

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All procedures performed in studies involving HUVEC were in accordance with the ethical standards of the institutional research committee (Bioethics Committee of the University of Patras) and with the 1964 Helsinki declaration and its later amendments or comparable ethical standards.

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Lamprou, M., Kastana, P., Kofina, F. et al. Pleiotrophin selectively binds to vascular endothelial growth factor receptor 2 and inhibits or stimulates cell migration depending on ανβ3 integrin expression. Angiogenesis 23, 621–636 (2020). https://doi.org/10.1007/s10456-020-09733-x

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