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Neuropilin-1 Targeting Photosensitization-Induced Early Stages of Thrombosis via Tissue Factor Release

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Abstract

Purpose

This article characterizes the vascular effects following vascular-targeted photodynamic therapy with a photosensitizer which actively targets endothelial cells.

Methods

This strategy was considered by coupling a chlorin to a heptapeptide targeting neuropilin-1 in human malignant glioma-bearing nude mice. A laser Doppler microvascular perfusion monitor was used to monitor microvascular blood perfusion in tumor tissue. Endothelial cells’ ultra structural integrity was observed by transmission electron microscopy. The consequences of photosensitization on tumor vessels, tissue factor expression, fibrinogen consumption, and thrombogenic effects were studied by immunohistochemical staining.

Results

Treatment of glioma-bearing mice with the conjugate showed a statistically significant tumor growth delay. Vascular effect was characterized by a decrease in tumor tissue blood flow at about 50% baseline during treatment not related to variations in temperature. This vascular shutdown was mediated by tumor blood vessels’ congestion. A pro-thrombotic behavior of targeted endothelial cells in the absence of ultra structural changes led to the induction of tissue factor expression from the earliest times post-treatment. Expression of tissue factor-initiated thrombi formation was also related to an increase in fibrinogen consumption.

Conclusion

Using a peptide-conjugated photosensitizer targeting neuropilin-1, induction of tissue factor expression immediately post-treatment, led to the establishment of thrombogenic effects within the vessel lumen.

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Abbreviations

Ahx:

6-aminohexanoic acid

a.i.:

arbitrary intensity

ATWLPPR:

H-Ala-Thr-Trp-Leu-Pro-Pro-Arg-OH

asHTF:

alternatively spiced human tissue factor

a.u.:

arbitrary units

BPD-MA:

benzoporphyrin derivative monoacid ring

BSA:

bovine serum albumine

DLI:

drug-light interval

FITC:

fluoresceine iso thio cyanate

HES:

hematoxylin, eosin and safran

HRP:

streptavidin-horseradish peroxidase

IL-6:

interleukin-6

i.v.:

intravenous

NRP-1:

neuropilin-1

PBS:

phosphate-buffered saline

PDT:

photodynamic therapy

PEG:

polyethylene glycol

PO2 :

oxygen pressure

ROS:

reactive oxygen species

RSM:

response surface methodology

s.d.:

standard deviation

TEM:

transmission electron microscopy

TEM buffer:

Tris EDTA molybdate buffer

TF:

tissue factor

TGD:

tumor growth delay

TNF-α:

tumor necrosis factor alpha

TPC:

5-(4-carboxyphenyl)-10,15,20-triphenylchlorin

V:

volume

VEGF:

vascular endothelial growth factor

VTP:

vascular targeted photodynamic therapy

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ACKNOWLEDGEMENTS

The authors would like to thank Denise Thiébaut, Marie Rouyer, and Carole Ramacci for their excellent technical assistance. We also thank Sophie Pinel and Anne-Sophie Chrétien for their collaboration. This work was supported by the research funds of the French “Ligue Nationale Contre le Cancer, Comités Lorrains.”

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Authors and Affiliations

  1. Centre de Recherche en Automatique de Nancy (CRAN), Nancy-University, CNRS, Centre Alexis Vautrin, Avenue de Bourgogne, Brabois, 54511, Vandœuvre-lès-Nancy, France

    Denise Bechet, Loraine Tirand, Thierry Bastogne, François Guillemin & Muriel Barberi-Heyob

  2. EA 4421 SiGReTO, Faculté de Médecine, Vandœuvre-lès-Nancy, France

    Béatrice Faivre & François Plénat

  3. Laboratoire d’Hématologie, Faculté de Pharmacie, Nancy-University, Nancy, France

    Béatrice Faivre

  4. Laboratoire d’Histopathologie Expérimentale et Moléculaire, Faculté de Médecine, Nancy-University, Nancy, France

    François Plénat & Corinne Bonnet

  5. Département de Chimie Physique des Réactions (DCPR), Nancy-University, CNRS, Nancy, France

    Céline Frochot

  6. GDR CNRS 3049 “Médicaments Photoactivables-Photochimiothérapie (PHOTOMED)”, Vandœuvre-lès-Nancy, France

    Thierry Bastogne, Céline Frochot, François Guillemin & Muriel Barberi-Heyob

Authors
  1. Denise Bechet
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  2. Loraine Tirand
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  6. Thierry Bastogne
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  7. Céline Frochot
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  8. François Guillemin
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  9. Muriel Barberi-Heyob
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Corresponding author

Correspondence to Muriel Barberi-Heyob.

Additional information

Loraine Tirand and Béatrice Faivre contributed equally to this study.

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Bechet, D., Tirand, L., Faivre, B. et al. Neuropilin-1 Targeting Photosensitization-Induced Early Stages of Thrombosis via Tissue Factor Release. Pharm Res 27, 468–479 (2010). https://doi.org/10.1007/s11095-009-0035-8

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  • DOI: https://doi.org/10.1007/s11095-009-0035-8

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