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Novel Carcinoembryonic-Antigen-(CEA)-Specific Pretargeting System to Assess Tumor Cell Viability after Irradiation of Colorectal Cancer Cells

Neues Carcinoembryonales-Antigen-(CEA-)Spezifisches Pretargeting-System zur Bestimmung der Tumorzellviabilität nach Bestrahlung kolorektaler Karzinomzellen

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Abstract

Purpose:

To date, no valid imaging modality exists for early response prediction to neoadjuvant radiochemotherapy in carcinoembryonic-antigen-(CEA)-expressing rectal cancers (UICC stages II and III). It is hypothesized that the uptake of an anti-CEA antibody is directly related to the number of viable tumor cells and may be quantified by immuno-positron emission tomography (immuno-PET). Therefore, we evaluated a novel pretargeting system using TF2, a humanized bispecific trivalent monoclonal antibody (mAb), directed against CEA and the IMP-288-peptide, a hapten for binding radiometals for imaging. Uptake and kinetics of the pretargeting system were investigated in vitro prior to and after irradiation.

Methods:

TF2 was labeled with 131I and IMP-288 with 111InCl3. The colorectal cancer cell lines HT29, SW480, and T84 with known varying CEA expression were incubated (≤72 hours) with 131I-TF2 or the TF2-111In-IMP-288 pretargeting system. Parallel cultures were irradiated with 2–10 Gy high-energy photons. Tracer uptake, proliferation, apoptosis, and CEA-RNA expression of cancer cells were investigated.

Results:

The uptake of tracers was dependent on CEA expression and cell count of the cell lines (uptake/106 cells: 0.3% in HT29, 1.5% in SW480, and 14% in T84, p < 0.001). The TF2-111In-IMP-288 pretargeting system showed a higher uptake after 4 and 72 hours compared to 131I-TF2 in parallel cultures. Only in one cell line (SW480) an increased apoptosis after irradiation could be detected. Irradiation increased dose dependently both the specific uptake of 131I-TF2 and of the TF2-111In-IMP-288 system (4-fold in HT29 and T84 after 10 Gy (72 hours), p < 0.001). These results were CEA-mRNA independent.

Conclusion:

This novel pretargeting system allows the quantitative analysis of CEA-expressing colorectal cancer cells and represents a promising tool for evaluation of tumor cell viability after irradiation.

Zusammenfassung

Hintergrund:

Derzeit gibt es beim CEA-exprimierenden Rektumkarzinom (der UICC-Stadien II und III) keine valide Diagnostik zur frühzeitigen Response-Beurteilung auf eine neoadjuvante Radiochemotherapie. Allerdings wird vermutet, daß mit einer Immuno-Positronen-Emissions-Tomographie (Immuno-PET) und einem anti-CEA-Antikörper die Anzahl viabler Tumorzellen nach erfolgter Radiatio quantifizierbar ist. Demzufolge wurde von unserer Arbeitsgruppe zur Etablierung eines Immuno-PETs ein neues Pretargeting-System evaluiert. Dieses Pretargeting-System besteht aus dem humanisierten bispezifischen trivalenten monoklonalen Antikörper (TF2), der an CEA sowie an das radioaktiv markierte Hapten-Peptid IMP-288 bindet. Wir untersuchten in vitro die Bindung und Kinetik dieses Pretargeting-Systems vor und nach Radiatio.

Methoden:

TF2 wurde mit 131I und IMP-288 mit 111InCl3 markiert. Die Inkubation (≤72 h) der kolorektalen Tumorzelllinien HT29, SW480 und T84 mit unterschiedlicher CEA-Expression erfolgte mit 131I-TF2 und dem TF2-111In-IMP-288-Pretargeting-System. Parallelkulturen wurden mit 2–10 Gy hochenergetischer Photonen bestrahlt. Neben der Bindung der markierten Substanzen wurden die Proliferation, Apoptose und CEA-RNA-Expression der Tumorzellen untersucht.

Ergebnisse:

Der Tracer-Uptake korrelierte sowohl mit der CEA-Expression als auch der Zellzahl der jeweiligen Zelllinie (Uptake/106 Zellen: 0,3% in HT29, 1,5% in SW480 und 14% in T84; p < 0,001). Das Pretargeting-System wurde nach 4 h und 72 h stärker gebunden als 131I-TF2 in Parallelkulturen. Lediglich in einer Zelllinie (SW480) konnte eine erhöhte Apoptoserate nach Bestrahlung nachgewiesen werden. Die Radiatio erhöhte dosisabhängig die spezifische Aufnahme des Pretargeting-Systems (4fach in HT29 und T84 nach 10 Gy (72 h), p < 0,001). Diese Ergebnisse waren CEA-mRNA-unabhängig.

Schlussfolgerung:

Das neue Pretargeting-System erlaubt eine quantitative Analyse der CEA-Expression kolorektaler Tumorzellen und ist eine vielversprechende Methode zur Evaluation der Tumorzellviabilität nach Radiatio.

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Authors and Affiliations

  1. Department of Nuclear Medicine, University Medical Center, Georg-August-University Göttingen, Göttingen, Germany

    Birgit Meller & Johannes Meller

  2. Department of Radiotherapy and Radiation Oncology, University Medical Center, Georg-August-University Göttingen, Göttingen, Germany

    Margarete Rave-Fränck

  3. Department of Radiology and Nuclear Medicine, University of Lübeck, Lübeck, Germany

    Christian Breunig

  4. Department of Clinical Pharmacology, University Medical Center, Georg-August-University Göttingen, Göttingen, Germany

    Markus Schirmer

  5. Department of Nuclear Medicine, Martin-Luther-University Halle, Halle, Germany

    Birgit Meller & Manfred Baehre

  6. Department of Radiotherapy, University of Lübeck, Lübeck, Germany

    Roger Nadrowitz

  7. Georg-August-University Göttingen, Göttingen, Germany

    Torsten Liersch

  8. Department of Nuclear Medicine, University Medical Center, Georg-August-University Göttingen, Robert-Koch-Str. 40, 37538, Göttingen, Germany

    Birgit Meller

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  1. Birgit Meller
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Meller, B., Rave-Fränck, M., Breunig, C. et al. Novel Carcinoembryonic-Antigen-(CEA)-Specific Pretargeting System to Assess Tumor Cell Viability after Irradiation of Colorectal Cancer Cells. Strahlenther Onkol 187, 120–126 (2011). https://doi.org/10.1007/s00066-010-2191-5

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  • DOI: https://doi.org/10.1007/s00066-010-2191-5

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