Nuklearmedizin 2019; 58(04): 309-318
DOI: 10.1055/a-0894-4127
Originalarbeit
© Georg Thieme Verlag KG Stuttgart · New York

Preclinical evaluation of peptide-based radiotracers for integrin αvβ6-positive pancreatic carcinoma

Präklinische Evaluation von Peptid-basierten Radiotracern für Integrin αvβ6-positive Pankreaskarzinome
Martin Müller
1   Clinical Cooperation Unit Nuclear Medicine, German Cancer Research Center (DKFZ) Heidelberg, Germany
2   Department of Nuclear Medicine, University Hospital Heidelberg, Germany
,
Annette Altmann
1   Clinical Cooperation Unit Nuclear Medicine, German Cancer Research Center (DKFZ) Heidelberg, Germany
2   Department of Nuclear Medicine, University Hospital Heidelberg, Germany
,
Max Sauter
2   Department of Nuclear Medicine, University Hospital Heidelberg, Germany
6   Department of Clinical Pharmacology and Pharmacoepidemiology, University Hospital Heidelberg, Germany
,
Thomas Lindner
2   Department of Nuclear Medicine, University Hospital Heidelberg, Germany
,
Dirk Jäger
3   Department of Medical Oncology, National Center for Tumor Diseases (NCT), University Hospital Heidelberg, Germany
,
Hendrik Rathke
1   Clinical Cooperation Unit Nuclear Medicine, German Cancer Research Center (DKFZ) Heidelberg, Germany
,
Christel Herold-Mende
4   Division of Experimental Neurosurgery, Department of Neurosurgery, University Hospital Heidelberg, Germany
,
Frederik Marmé
5   Department of Gynecologic Oncology, National Center for Tumor Diseases (NCT) and Department of Obstetrics and Gynecology, University Women’s Clinic, University Hospital Heidelberg, Germany
,
John Babich
7   Department for Radiology, Weill Cornell Medicine, New York, NY, USA
,
Walter Mier
2   Department of Nuclear Medicine, University Hospital Heidelberg, Germany
,
Uwe Haberkorn
1   Clinical Cooperation Unit Nuclear Medicine, German Cancer Research Center (DKFZ) Heidelberg, Germany
2   Department of Nuclear Medicine, University Hospital Heidelberg, Germany
8   Translational Lung Research Center Heidelberg (TLRC), German Center for Lung Research (DZL), Heidelberg, Germany
› Author Affiliations
Further Information

Publication History

30 October 2018

12 April 2019

Publication Date:
10 May 2019 (online)

Abstract

Introduction Integrin αvβ6 shows a high expression rate in several cancer entities. As it is absent in most healthy adult tissues, it represents a promising target for tumor targeting with peptidic radiotracers. This study was performed to pave the way of the recently published αvβ6-binding peptide SFLAP3 for the clinical application in patients with pancreatic cancer.

Methods The expression of integrin αvβ6 on several pancreatic cancer cell lines was assessed using flow cytometry and cell binding assays. The affinity was determined in competition binding assays followed by internalization and efflux studies. To increase the affinity, the binding sequence was modified and trimerization of the SFLAP3 peptide was achieved by oxime ligation. PET and biodistribution assays were conducted in Capan-2 tumor bearing mice. Finally, a first pancreatic tumor patient was examined with 68Ga-DOTA-SFLAP3.

Results Flow cytometric analysis and in vitro cell binding revealed high expression of integrin αvβ6 on most pancreatic tumor cell lines. Modification of SFLAP3 led to compounds with improved in vitro binding properties. Unfortunately, these superior properties could not be transferred into improved pharmacokinetics. Consequently, the first pancreatic tumor patient was examined with 68Ga-DOTA-SFLAP3. The PET revealed specific accumulation (with SUV(max) values in the metastases ranging from 5 to 10) and a long retention in the tumor.

Conclusion SFLAP3 showed high affinity to integrin αvβ6 on pancreatic cancer cell lines. The in vitro performance could be confirmed in tumor bearing mice and by PET imaging. These data suggest that DOTA-SFLAP3 is a promising tracer for targeting αvβ6-expressing pancreatic tumors.

Zusammenfassung

Einleitung Die Expressionsrate von Integrin αvβ6 ist in vielen Krebsarten stark erhöht. Da es in gesunden adulten Geweben fast nicht vorhanden ist, stellt es ein vielversprechendes Zielprotein für zielgerichtete Krebsdiagnostik mit peptidischen Radiotracern dar. Diese Arbeit wurde durchgeführt, um den Weg des kürzlich veröffentlichten αvβ6-bindenden Peptids SFLAP3 für die klinische Anwendung bei Patienten mit Pankreaskarzinom zu ebnen.

Methoden Die Integrin αvβ6-Expression verschiedener Pankreaskarzinomzelllinien wurde mittels Durchflusszytometrie und Zellbindungsassays untersucht. Die Affinität wurde in kompetitiven Bindungsassays bestimmt und Internalisierungs- und Efflux-Studien durchgeführt. Um die Affinität zu erhöhen, wurde die Bindungssequenz modifiziert und das SFLAP3-Peptid durch Oximligation trimerisiert. PET- und Bioverteilungsversuche wurden in Capan-2-tumortragenden Mäusen durchgeführt. Letztendlich wurde ein erster Pankreas-Tumor-Patient mit 68Ga-DOTA-SFLAP3 untersucht.

Ergebnisse Die durchflusszytometrische Analyse und die in vitro-Zellbindungen zeigten auf den meisten Pankreas-Tumorzelllinien, eine hohe Expression von Integrin αvβ6. Modifikationen von SFLAP3 führten zu Verbindungen mit verbesserten in vitro-Bindungseigenschaften. Leider führten diese Verbesserungen im Tiermodell zu keiner verbesserten Pharmakokinetik. Folglich wurde für die Untersuchung eines ersten Pankreas-Tumor-Patienten SFLAP3, das Peptid mit den ausgewogensten Eigenschaften, ausgewählt. Die PET-Studie mit 68Ga-DOTA-SFLAP3 zeigte eine spezifische Tracerakkumulation (mit SUV(max)-Werten in den Metastasen im Bereich von 5 bis 10) mit langer Tumor-Retention.

Schlussfolgerung SFLAP3 zeigte eine hohe Affinität zu Integrin αvβ6 auf Pankreas-Tumorzelllinien. Die in vitro Eigenschaften konnten in tumortragenden Mäusen und durch PET-Bildgebung eines Pankreaskarzinom-Patienten bestätigt werden. Diese Daten legen nahe, dass DOTA-SFLAP3 ein vielversprechender Tracer für αvβ6-exprimierende Pankreastumore ist.

Supplementary material

 
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