In vivo imaging of immunotoxin treatment using Katushka-transfected A-431 cells in a murine xenograft tumour model
| Autor: | Rainer Fischer, Michael Stöcker, Alessa Pardo, Theo Thepen, Florian Kampmeier, Georg Melmer, Stefan Barth |
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| Rok vydání: | 2012 |
| Předmět: |
Cancer Research
Fluorescence-lifetime imaging microscopy Skin Neoplasms Virulence Factors medicine.medical_treatment Bacterial Toxins Immunology Exotoxins Mice Nude Antineoplastic Agents Biology Transfection Mice In vivo Immunotoxin Cell Line Tumor medicine Animals Humans Immunology and Allergy Doxorubicin Epidermal growth factor receptor ADP Ribose Transferases Mice Inbred BALB C Spectroscopy Near-Infrared Immunotoxins Carcinoma Immunotherapy Xenograft Model Antitumor Assays Molecular biology Molecular Imaging ErbB Receptors Disease Models Animal Luminescent Proteins Oncology biology.protein Female Preclinical imaging medicine.drug |
| Zdroj: | Cancer Immunology, Immunotherapy. 61:1617-1626 |
| ISSN: | 1432-0851 0340-7004 |
| Popis: | Preclinical in vivo analyses of treatment responses are an important prerequisite to evaluate new therapeutics. Molecular in vivo imaging in the far red (FR)/near infra red (NIR) is a promising method, as it enables measurements at different time points in individual animals, thereby reducing the number of animals required, while increasing statistical significance. Here, we show the establishment of a method to monitor response to treatment using fluorescent cells, expressing the epidermal growth factor receptor (EGFR), a target already used in therapy.We transfected A-431 tumour cells with the far red-emitting protein Katushka (Kat2), resulting in strong fluorescence allowing for the monitoring of tumour growth when implanted in BALB/c nu/nu mice with a CRi Maestro in vivo imager. We targeted A-431 cells with a previously reported immunotoxin (IT), consisting of the anti-EGFR antibody single-chain variable fragment (scFv) 425, fused to Pseudomonas aeruginosa Exotoxin A' (ETA'). In addition, EGFR expression was verified using the 425(scFv) conjugated to a NIR dye BG-747 through a SNAP-tag linker.The results show the feasibility to evaluate response to treatment in vivo by FR imaging, while at the same location detecting EGFR expression. Treatment with 425(scFv)-ETA' resulted in decelerated tumour growth, while not affecting the overall health of the animals. This is in contrast to treatment with Doxorubicin, which, although decreasing the tumour size, resulted in poor health.We developed a novel method to non-invasively determine treatment responses by in vivo imaging of multiple parameters which showed the efficacy of 425(scFv)-ETA'. |
| Databáze: | OpenAIRE |
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