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Novel PSCA targeting scFv-fusion proteins for diagnosis and immunotherapy of prostate cancer

 
: Kessler, C.; Pardo, A.; Tur, M.K.; Gattenlöhner, S.; Fischer, R.; Kolberg, K.; Barth, S.

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Journal of cancer research and clinical oncology 143 (2017), No.10, pp.2025-2038
ISSN: 0171-5216 (Print)
ISSN: 0084-5353
ISSN: 0943-9382
ISSN: 1432-1335 (Online)
English
Journal Article
Fraunhofer IME ()

Abstract
Purpose Despite great progress in the diagnosis and treatment of localized prostate cancer (PCa), there remains a need for new diagnostic markers that can accurately distinguish indolent and aggressive variants. One promising approach is the antibody-based targeting of prostate stem cell antigen (PSCA), which is frequently overexpressed in PCa. Here, we show the construction of a molecular imaging probe comprising a humanized scFv fragment recognizing PSCA genetically fused to an engineered version of the human DNA repair enzyme O6-alkylguanine-DNA alkyltransferase (AGT), the SNAP-tag, enabling specific covalent coupling to various fluorophores for diagnosis of PCa. Furthermore, the recombinant immunotoxin (IT) PSCA(scFv)-ETA' comprising the PSCA(scFv) and atruncated version of Pseudomonas exotoxin A (PE, ETA') was generated. Methods We analyzed the specific binding and internalization behavior of the molecular imaging probe PSCA(scFv)SNAP in vitro by flow cytometry and live cell imaging, compared to the corresponding IT PSCA(scFv)-ETA'. The cytotoxic activity of PSCA(scFv)-ETA' was tested using cell viability assays. Specific binding was confirmed on formalin-fixed paraffin-embedded tissue specimen of early and advanced PCa. Results Alexa -Fluor (R) 647 labeling of PSCA(scFv)-SNAP confirmed selective binding to PSCA, leading to rapid internalization into the target cells. The recombinant IT PSCA(scFv)-ETA' showed selective binding leading to internalization and efficient elimination of target cells. Conclusions Our data demonstrate, for the first time, the specific binding, internalization, and cytotoxicity of a scFv-based fusion protein targeting PSCA. Immunohistochemical staining confirmed the specific ex vivo binding to primary PCa material.

: http://publica.fraunhofer.de/documents/N-480450.html