Systemic treatment with 4-²¹¹At - phenylalanine enhances survival of rats with intracranial glioblastoma

Summary

Objective: Increased amino acid transport in brain tumours is used for diagnostic purposes. It has been shown that the α-emitting radionuclide astatine-211 labeled to L-phenylalanine is taken up by glioblastoma cells. We here tested, if systemic treatment with 4-[²¹¹At] astatine-phenylalanine (At-Phe) has a beneficial effect on survival of rats with intracranial glioblastoma. Animals, methods: The rat glioblastoma cell line BT4Ca was implanted into the prefrontal cortex of female BDIX rats by stereotaxic microinjection (10 000 cells/3 μl; n = 83). 3 days after implantation At-Phe or phosphate buffered saline were injected intravenously. A third group was treated twice, i.e., on day 3 and 10. Health condition was assessed each day by using a score system. Rats were sacrificed on days 6, 10, 13 and 17 after implantation, or when showing premortal health condition to measure tumour volume and necrosis. The proliferation index (PI) was assessed after immunohistochemical staining of Ki-67. Results: Survival time of rats treated twice with At-Phe was significantly prolonged. Additionally, both At-Phe-treated groups remained significantly longer in a better health condition. Rats with poor health status had larger tumours than rats with fair health condition. Overall, irrespective of treatment the PI was reduced in rats with poor health condition. Necrosis was larger in rats treated twice with At-Phe. Conclusion: Intravenous treatment with At-Phe enhanced survival time of rats with intracranial glioblastomas and improved health condition. These results encourage studies using local treatment of intracranial glioblastoma with At-Phe, either by repeated local injection or by intracavital application after tumour resection.

Zusammenfassung

Ziel: Die erhöhte Aufnahme von Aminosäuren in Hirntumoren wird für diagnostische Zwecke genutzt. Es wurde bereits gezeigt, dass mit dem Alpha-Strahler Astat-211 markiertes L-Phenylalanin ebenfalls in Glioblastom zellen aufgenommen wird. Hier wurde getestet, ob die systemische Gabe von 4-[²¹¹At|Astat-Phenylalanin (²¹¹At-Phe) einen positiven Effekt auf das Überleben von Ratten mit intrakranial implantierten Glioblastomen hat. Tiere, Methodik: Durch stereotaktische Mikroinjektion wurde die Ratten-Glioblastom-Zelllinie BT4Ca (10 000 Zellen/3 μl) in den präfrontalen Kortex weiblicher BDIX-Ratten implantiert. Drei Tage nach der Implantation wurde ²¹¹At-Phe oder Phosphat-gepufferte Kochsalz- Lösung intravenös appliziert. Eine dritte Gruppe wurde ein zweites Mal behandelt, d. h. am 3. Tag und 10. Tag. Der Gesundheitszustand wurde täglich in einem Punkte-System erfasst. Zur Untersuchung des Tumorvolumens und der Tumornekrose wurden Ratten an den Tagen 6, 10, 13 und 17 nach der Implantation getötet, oder wenn ihr Gesundheitszustand ein weiteres Überleben unverantwortlich erscheinen ließ. Der Proliferationsindex (PI) wurde nach immunhistochemischer Färbung mit Ki-67 bestimmt. Ergebnisse: Das Überleben der Ratten nach zweimaliger Applikation von ²¹¹At-Phe war signifikant verlängert. Beide mit ²¹¹At-Phe behandelten Gruppen verblieben signifikant länger in einem besseren Gesundheitszustand. Die Ratten mit einem schlechten Gesundheitszustand hatten größere Tumoren als die mit gutem Gesundheitszustand. Der PI war unabhängig vom Behandlungsweg geringer in Tieren mit schlechtem Gesundheitszustand. Der Nekroseanteil in den Tumoren von zweifach mit ²¹¹At-Phe behandelten Tieren war größer. Schlussfolgerung: Intravenöse Applikation von ²¹¹At-Phe verlängert und verbessert das Überleben von Ratten mit intrakranial implantierten Glioblastomen. Die Ergebnisse legen weitere Versuche nahe, direkte lokale Applikationen von ²¹¹At-Phe an intrakranial implantierten Glioblastomen vorzunehmen, oder intrakavitäre Applikationen nach Tumor- Resektion.

Keywords

Glioblastoma - intracranial glioblastoma rat model - astatine-211 - At-phenylalanine

Schlüsselwörter

Glioblastom - intrakraniales Glioblastom- Rattenmodell - Astat-211 - At-Phenylalanin

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