Selective targeting and potent control of tumor growth using an EphA2/CD3-Bispecific single-chain antibody construct.
The EphA2 receptor tyrosine kinase is frequently overexpressed and functionally altered in malignant cells and thus provides opportunities for selective targeting of tumor cells. We describe here the development of a novel, bispecific single-chain antibody (bscAb) referred to as bscEphA2xCD3. This molecule simultaneously targets EphA2 on tumor cells and the T-cell receptor/CD3 complex on T cells and possesses structural and functional characteristics of the recently developed BiTE technology. An EphA2-specific single-chain antibody was selected for recognition of an epitope that is preferentially exposed on malignant cells based on the concept of epitope exclusion; this was fused to a CD3-specific single-chain antibody to generate bscEphA2xCD3. The resultant bscAb redirected unstimulated human T cells to lyse EphA2-expressing tumor cells both in vitro and in vivo. In separate experiments, efficient tumor cell lysis was achieved in vitro at drug concentrations <or=1 microg/mL, at a low T-cell effector-to-tumor target cell ratio (1:1), and with tumor cells that possess few available binding sites (2,400 per cell) for bscEphA2xCD3. Time-lapsed microscopy revealed potent cytotoxic activity of bscEphA2xCD3-activated T cells against monolayers of malignant cells but not against monolayers of nontransformed EphA2-positive cells except at the edges of the monolayer where the target epitope was exposed. BscEphA2xCD3 was also efficacious in human xenograft mouse models modified to show human T-cell killing of tumors. Together, our results reveal opportunities for redirecting the potent activity of cytotoxic T cells towards tumor cells that express selectively accessible epitopes and establish EphA2-specific bscAb molecules as novel and potent therapeutics with selectivity for tumor cells.
Hammond
Scott A
SA
Medimmune, Inc., Gaithersburg, Maryland 20878, USA. hammonds@medimmune.com
Lutterbuese
Ralf
R
Roff
Shannon
S
Lutterbuese
Petra
P
Schlereth
Bernd
B
Bruckheimer
Elizabeth
E
Kinch
Michael S
MS
Coats
Steve
S
Baeuerle
Patrick A
PA
Kufer
Peter
P
Kiener
Peter A
PA
D000818:Animals; D018033:Antibodies, Bispecific / Q000276:immunology / Q000494:pharmacology*; D000918:Antibody Specificity; D017252:CD3 Complex / Q000276:immunology*; D016466:CHO Cells; D048708:Cell Growth Processes / Q000276:immunology; D045744:Cell Line, Tumor; D006224:Cricetinae; D003412:Cricetulus; D000939:Epitopes / Q000276:immunology; D006801:Humans; D007116:Immunization, Passive / Q000379:methods*; D007365:Intercellular Junctions / Q000276:immunology; D008213:Lymphocyte Activation; D051379:Mice; D016688:Mice, Inbred NOD; D016513:Mice, SCID; D009369:Neoplasms / Q000276:immunology / Q000473:pathology / Q000628:therapy*; D036104:Receptor, EphA2 / Q000276:immunology*; D013601:T-Lymphocytes / Q000276:immunology; D023041:Xenograft Model Antitumor Assays
D018033:Antibodies, Bispecific; D017252:CD3 Complex; D000939:Epitopes; D036104:Receptor, EphA2