Abstract 2806: Development of an iRFP713-based orthotopic patient derived brain cancer model to study tumor development in vivo
Authors: Lashuk Kanstantsin , Bersano Jacqueline , Lenhard Dorothee , Oswald Eva , Klingner Kerstin , Schüler Julia 
Published: July 2021

While the use of bioluminescent proteins for molecular imaging is a powerful technology to support our knowledge of complex processes, with the advent of near infrared-labeling capabilities optical imaging can be pursuit with deeper tissue penetration, a better signal to background ratio and a quicker image acquisition without the need to inject substrates in preclinical oncology mouse models. This is specifically important in orthoptic brain tumors, when the substrate concentration in the tumor tissue might be influenced by the blood-brain barrier. In our recent study, we validated the feasibility of stable or transient transfection of human brain tumor cell lines and patient derived xenografts to determine tumor load in an intracranial setting. As a proof-of principle, we stably transfected the human glioblastoma cell line U87MG with an iRFP713 construct and followed tumor growth of intracranially implanted NSG mice. As a follow-up, five PDX of pediatric brain tumors (two Ependymoma, two Medulloblastoma, one high grade Glioblastoma) were transiently transfected with the same construct 24h before intracranial implantation. Tumor load was determined twice a week with the Pearl trilogy system (LiCor, Germany) and animals were examined for neurological symptoms every day. When the overall condition of the mice got deteriorated, animals were taken down and brains fixed in formalin. The tumor load and localization was confirmed by H&E and human-specific IHC for LamininB. The comparison between non-transfected and transfected lines of one specific tumor model, revealed that the tumor growth was very similar in both settings. The tumor take rate was as well not affected. In the subsequent analyses we could show that the tumor load linearly correlated with the imaging signal. Moreover, the iRFP713 signal was stable post-resection of the tumor. Thus, it was possible to correlate the tumor load quantification via iRFP713-Protein expression and LamininB-IHC within one slide. The transient transfection was successful for the five tested models. When termination criteria were reached, over 90% of the tumor tissue were expressing the iRFP713 protein. The more general applicability for larger cohorts of mice (&gt; 20) and drug testing experiments will be evaluated soon. We believe the superior optical properties of iRFP713 will be a valuable asset to overcome some of the complications inherent to imaging live animals and a powerful tool for preclinical drug development in clinical relevant mouse models of brain cancer.

Citation Format: Kanstantsin Lashuk, Jacqueline Bersano, Dorothee Lenhard, Eva Oswald, Kerstin Klingner, Julia Schüler. Development of an iRFP713-based orthotopic patient derived brain cancer model to study tumor development in vivo [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr 2806.