A bispecific diabody directed against prostate-specific membrane antigen and CD3 induces T-cell mediated lysis of prostate cancer cells

Abstract

Background Although cancer of the prostate is one of the most commonly diagnosed cancers in men, no curative treatment currently exists after its progression beyond resectable boundaries. Therefore, new agents for targeted treatment strategies are needed. Cross-linking of tumor antigens with T-cell associated antigens by bispeciWc monoclonal antibodies have been shown to increase antigen-speciWc cytotoxicity in T-cells. Since the prostate-speciWc membrane antigen (PSMA) represents an excellent tumor target, immunotherapy with bispeciWc diabodies could be a promising novel treatment option for prostate cancer. Methods A heterodimeric diabody speciWc for human PSMA and the T-cell antigen CD3 was constructed from the DNA of anti-CD3 and anti-PSMA single chain Fv fragments (scFv). It was expressed in E. coli using a vector containing a bicistronic operon for co-secretion of the hybrid scFv V H CD3-V L PSMA and V H PSMA-V L CD3. The resulting PSMAxCD3 diabody was puriWed from the periplasmic extract by immobilized metal aYnity chromatography (IMAC). The binding properties were tested on PSMA-expressing prostate cancer cells and PSMA-negative cell lines as well as on Jurkat cells by Xow cytometry.

MOR209/ES414, a Novel Bispecific Antibody Targeting PSMA for the Treatment of Metastatic Castration-Resistant Prostate Cancer

“…The ability of MOR209/ES414 to prevent tumor growth in vivo was evaluated in two xenograft models of prostate cancer (C4-2B and MDA-PCa-2b) in NOD/SCID mice using adoptively transferred human T cells, similar to previous studies using bispecific antibody fragments (15,16). The ability of MOR209/ES414 to prevent growth of C4-2B tumors was compared with the tandem scFv control.…”

“…PSMA has been used as a target for the development of diagnostics (10) and for therapeutic mAbs (11)(12)(13)(14). Bispecific antibody fragments in either a diabody format (15) or a tandem singlechain variable fragment (scFv) format (16) targeting PSMA and the T-cell receptor (TCR) complex subunit CD3e have been described previously. One of these molecules, BAY2010112, is currently under early clinical evaluation.…”

Redirected T-Cell Killing of Solid Cancers Targeted with an Anti-CD3/Trop-2–Bispecific Antibody Is Enhanced in Combination with Interferon-α

“…1). The potential of this modality has been demonstrated preclinically for both hematopoietic (2,3) and solid cancers (4)(5)(6)(7)(8). Tandem scFvs consisting of an anti-CD3 and an anti-TAA domain, termed bispecific T-cell engagers (BiTE), have advanced the furthest, with two agents under clinical investigation (9).…”

Highly Enhanced Cytotoxicity of a Dimeric Bispecific Diabody, the hEx3 Tetrabody

“…VH A -VL B and VH B -VL A ) with a glycine-rich linker (GGGGS; 8,24). The hetero scFvs can also form higher multimeric structures (25), and the multimeric bispecific diabodies formed are expected to * This work was supported by grants-in-aid for scientific research from the Ministry of Education, Science, Sports, and Culture of Japan (to R. A. and I. K.) and by grants from the New Energy and Industrial Technology Development Organization (NEDO) of Japan. This work was also supported by the Program for Promotion of Fundamental Studies in Health Sciences of the National Institute of Biomedical Innovation.…”