Heterodimerization of T cell engaging bispecific antibodies to enhance specificity against pancreatic ductal adenocarcinoma Journal Article
| Authors: | Long, A. W.; Xu, H.; Santich, B. H.; Guo, H.; Hoseini, S. S.; de Stanchina, E.; Cheung, N. K. V. |
| Article Title: | Heterodimerization of T cell engaging bispecific antibodies to enhance specificity against pancreatic ductal adenocarcinoma |
| Abstract: | Background: EGFR and/or HER2 expression in pancreatic cancers is correlated with poor prognoses. We generated homodimeric (EGFRxEGFR or HER2xHER2) and heterodimeric (EGFRxHER2) T cell-engaging bispecific antibodies (T-BsAbs) to direct polyclonal T cells to these antigens on pancreatic tumors. Methods: EGFR and HER2 T-BsAbs were constructed using the 2 + 2 IgG-[L]-scFv T-BsAbs format bearing two anti-CD3 scFvs attached to the light chains of an IgG to engage T cells while retaining bivalent binding to tumor antigens with both Fab arms. A Fab arm exchange strategy was used to generate EGFRxHER2 heterodimeric T-BsAb carrying one Fab specific for EGFR and one for HER2. EGFR and HER2 T-BsAbs were also heterodimerized with a CD33 control T-BsAb to generate ‘tumor-monovalent’ EGFRxCD33 and HER2xCD33 T-BsAbs. T-BsAb avidity for tumor cells was studied by flow cytometry, cytotoxicity by T-cell mediated 51Chromium release, and in vivo efficacy against cell line-derived xenografts (CDX) or patient-derived xenografts (PDX). Tumor infiltration by T cells transduced with luciferase reporter was quantified by bioluminescence. Results: The EGFRxEGFR, HER2xHER2, and EGFRxHER2 T-BsAbs demonstrated high avidity and T cell-mediated cytotoxicity against human pancreatic ductal adenocarcinoma (PDAC) cell lines in vitro with EC50s in the picomolar range (0.17pM to 18pM). They were highly efficient in driving human polyclonal T cells into subcutaneous PDAC xenografts and mediated potent T cell-mediated anti-tumor effects. Both EGFRxCD33 and HER2xCD33 tumor-monovalent T-BsAbs displayed substantially reduced avidity by SPR when compared to homodimeric EGFRxEGFR or HER2xHER2 T-BsAbs (∼150-fold and ∼6000-fold respectively), tumor binding by FACS (8.0-fold and 63.6-fold), and T-cell mediated cytotoxicity (7.7-fold and 47.2-fold), while showing no efficacy against CDX or PDX. However, if either EGFR or HER2 was removed from SW1990 by CRISPR-mediated knockout, the in vivo efficacy of heterodimeric EGFRxHER2 T-BsAb was lost. Conclusion: EGFR and HER2 were useful targets for driving T cell infiltration and tumor ablation. Two arm Fab binding to either one or both targets was critical for robust anti-tumor effect in vivo. By engaging both targets, EGFRxHER2 heterodimeric T-BsAb exhibited potent anti-tumor effects if CDX or PDX were EGFR+HER2+ double-positive with the potential to spare single-positive normal tissue. © The Author(s) 2024. |
| Keywords: | cancer survival; controlled study; human cell; area under the curve; nonhuman; flow cytometry; antigen expression; t lymphocyte; animal cell; mouse; animal tissue; epidermal growth factor receptor; epidermal growth factor receptor 2; animal experiment; animal model; in vivo study; antineoplastic activity; tumor regression; tumor xenograft; cetuximab; tumor antigen; drug distribution; antibody specificity; egfr; cancer control; lymphocytic infiltration; trastuzumab; antigen binding; heterodimer; fluorescence activated cell sorting; bioluminescence; immunoglobulin g antibody; t lymphocyte activation; single chain fragment variable antibody; antibody affinity; cd33 antigen; cell homing; her2; cell mediated cytotoxicity; tumor microenvironment; immunoglobulin f(ab) fragment; pancreatic ductal adenocarcinoma; pancreatic ductal carcinoma; heterodimerization; cd3 antibody; t lymphocyte antibody; bispecific antibody; chromium 51; human; article; luciferase assay; clustered regularly interspaced short palindromic repeat; ec50; bxpc-3 cell line; chromium release assay; sw1990 cell line |
| Journal Title: | Journal of Hematology & Oncology |
| Volume: | 17 |
| ISSN: | 1756-8722 |
| Publisher: | Biomed Central Ltd |
| Date Published: | 2024-04-23 |
| Start Page: | 20 |
| Language: | English |
| DOI: | 10.1186/s13045-024-01538-5 |
| PROVIDER: | scopus |
| PMCID: | PMC11036555 |
| PUBMED: | 38650005 |
| DOI/URL: | |
| Notes: | Article -- MSK Cancer Center Support Grant (P30 CA008748) acknowledged in PubMed and PDF -- MSK corresponding author is Nai-Kong V. Cheung -- Source: Scopus |
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