Concurrent binding of anti-EphA3 antibody and ephrin-A5 amplifies EphA3 signaling and downstream responses: Potential as EphA3-specific tumor-targeting reagents


Autoria(s): Vearing, Christopher; Lee, Fook-Thean; Wimmer-Kleikamp, Sabine; Spirkoska, Violeta; To, Catherine; Stylianou, Con; Spanevello, Mark; Brechbiel, Martin; Boyd, Andrew W.; Scott, Andrew M.; Lackmann, Martin
Data(s)

01/01/2005

Resumo

The Eph receptor tyrosine kinases and their membrane-bound ephrin ligands form a unique cell-cell contact-mediated system for controlling cell localization and organization. Their high expression in a wide variety of human tumors indicates a role in tumor progression, and relatively low Eph and ephrin levels in normal tissues make these proteins potential targets for anticancer therapies. The monoclonal antibody IIIA4, previously used to isolate EphA3, binds with subnanomolar affinity to a conformation-specific epitope within the ephrin-binding domain that is closely adjacent to the low-affinity ephrin-A5 heterotetramerization site. We show that similar to ephrin-A5, preclustered IIIA4 effectively triggers EphA3 activation, contraction of the cytoskeleton, and cell rounding. BIAcore analysis, immunoblot, and confocal microscopy of wild-type and mutant EphA3 with compromised ephrin-A5 or IIIA4-binding capacities indicate that IIIA4 binding triggers an EphA3 conformation which is permissive for the assembly of EphA3/ephrin-A5-type signaling clusters. Furthermore, unclustered IIIA4 and ephrin-A5 Fc applied in combination initiate greatly enhanced EphA3 signaling. Radiometal conjugates of ephrin-A5 and IIIA4 retain their affinity, and in mouse xenografts localize to, and are internalized rapidly into EphA3-positive, human tumors. These findings show the biological importance of EphA3/ ephrin-A5 interactions and that ephrin-A5 and IIIA4 have great potential as tumor targeting reagents.

Identificador

http://espace.library.uq.edu.au/view/UQ:75021

Idioma(s)

eng

Publicador

American Association for Cancer Research

Palavras-Chave #Oncology #Receptor Tyrosine Kinases #Diverse Roles #Ligand #Cells #Carcinogenesis #Embryogenesis #Inhibition #Attachment #Ephb2 #Hek #C1 #321015 Oncology and Carcinogenesis #730108 Cancer and related disorders #11 Medical and Health Sciences
Tipo

Journal Article