Format-tuning of in vivo-launched bispecific T cell engager enhances efficacy against renal cell carcinoma

Background Advanced clear cell renal cell carcinoma (ccRCC) is a prevalent kidney cancer for which long-term survival rates are abysmal, though immunotherapies are showing potential. Not yet clinically vetted are bispecific T cell engagers (BTEs) that activate T cell-mediated cancer killing through...

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Main Authors: Carl H June, David Weiner, Christopher A Chuckran, Daniel Park, Ryan P O’Connell, Kevin Liaw, Nils Wellhausen, Pratik S Bhojnagarwala, Devivasha Bordoloi, Nicholas Shupin, Daniel Kulp
Format: Article
Language:English
Published: BMJ Publishing Group 2024-06-01
Series:Journal for ImmunoTherapy of Cancer
Online Access:https://jitc.bmj.com/content/12/6/e008733.full
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author Carl H June
David Weiner
Christopher A Chuckran
Daniel Park
Ryan P O’Connell
Kevin Liaw
Nils Wellhausen
Pratik S Bhojnagarwala
Devivasha Bordoloi
Nicholas Shupin
Daniel Kulp
author_facet Carl H June
David Weiner
Christopher A Chuckran
Daniel Park
Ryan P O’Connell
Kevin Liaw
Nils Wellhausen
Pratik S Bhojnagarwala
Devivasha Bordoloi
Nicholas Shupin
Daniel Kulp
author_sort Carl H June
collection DOAJ
description Background Advanced clear cell renal cell carcinoma (ccRCC) is a prevalent kidney cancer for which long-term survival rates are abysmal, though immunotherapies are showing potential. Not yet clinically vetted are bispecific T cell engagers (BTEs) that activate T cell-mediated cancer killing through intercellular synapsing. Multiple BTE formats exist, however, with limited cross-characterizations to help optimize new drug design. Here, we developed BTEs to treat ccRCC by targeting carbonic anhydrase 9 (CA9) while characterizing the persistent BTE (PBTE) format and comparing it to a new format, the persistent multivalent T cell engager (PMTE). These antibody therapies against ccRCC are developed as both recombinant and synthetic DNA (synDNA) medicines.Methods Antibody formatting effects on binding kinetics were assessed by flow cytometry and intercellular synaptic strength assays while potency was tested using T-cell activation and cytotoxicity assays. Mouse models were used to study antibody plasma and tumor pharmacokinetics, as well as antitumor efficacy as both recombinant and synDNA medicines. Specifically, three models using ccRCC cell line xenografts and human donor T cells in immunodeficient mice were used to support this study.Results Compared with a first-generation BTE, we show that the PBTE reduced avidity, intercellular synaptic strength, cytotoxic potency by as much as 33-fold, and ultimately efficacy against ccRCC tumors in vivo. However, compared with the PBTE, we demonstrate that the PMTE improved cell avidity, restored intercellular synapses, augmented cytotoxic potency by 40-fold, improved tumor distribution pharmacokinetics by 2-fold, and recovered synDNA efficacy in mouse tumor models by 20-fold. All the while, the PMTE displayed a desirable half-life of 4 days in mice compared with the conventional BTE’s 2 hours.Conclusions With impressive efficacy, the CA9-targeted PMTE is a promising new therapy for advanced ccRCC, which can be effectively delivered through synDNA. The highly potent PMTE format itself is a promising new tool for future applications in the multispecific antibody space.
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spelling doaj-art-e668ca4eaa1e4321a85a2c4dec2793ca2025-08-20T03:07:28ZengBMJ Publishing GroupJournal for ImmunoTherapy of Cancer2051-14262024-06-0112610.1136/jitc-2023-008733Format-tuning of in vivo-launched bispecific T cell engager enhances efficacy against renal cell carcinomaCarl H June0David Weiner1Christopher A Chuckran2Daniel Park3Ryan P O’Connell4Kevin Liaw5Nils Wellhausen6Pratik S Bhojnagarwala7Devivasha Bordoloi8Nicholas Shupin9Daniel Kulp102Department of Medicine, Perelman School of Medicine at the University of Pennsylvania, Philadelphia, PA, USAThe Wistar Institute, Philadelphia, PA, USA4 LUMICKS, Waltham, Massachusetts, USA2 Vaccine & Immunotherapy Center, Wistar Institute, Philadelphia, Pennsylvania, USA1 University of Pennsylvania Perelman School of Medicine, Philadelphia, Pennsylvania, USAVaccine and Immunotherapy Center, Wistar Institute, Philadelphia, Pennsylvania, USA3 Center for Cellular Immunotherapies, University of Pennsylvania Perelman School of Medicine, Philadelphia, Pennsylvania, USAVaccine and Immunotherapy Center, Wistar Institute, Philadelphia, Pennsylvania, USAVaccine and Immunotherapy Center, Wistar Institute, Philadelphia, Pennsylvania, USA2 Vaccine & Immunotherapy Center, Wistar Institute, Philadelphia, Pennsylvania, USA2 Vaccine & Immunotherapy Center, Wistar Institute, Philadelphia, Pennsylvania, USABackground Advanced clear cell renal cell carcinoma (ccRCC) is a prevalent kidney cancer for which long-term survival rates are abysmal, though immunotherapies are showing potential. Not yet clinically vetted are bispecific T cell engagers (BTEs) that activate T cell-mediated cancer killing through intercellular synapsing. Multiple BTE formats exist, however, with limited cross-characterizations to help optimize new drug design. Here, we developed BTEs to treat ccRCC by targeting carbonic anhydrase 9 (CA9) while characterizing the persistent BTE (PBTE) format and comparing it to a new format, the persistent multivalent T cell engager (PMTE). These antibody therapies against ccRCC are developed as both recombinant and synthetic DNA (synDNA) medicines.Methods Antibody formatting effects on binding kinetics were assessed by flow cytometry and intercellular synaptic strength assays while potency was tested using T-cell activation and cytotoxicity assays. Mouse models were used to study antibody plasma and tumor pharmacokinetics, as well as antitumor efficacy as both recombinant and synDNA medicines. Specifically, three models using ccRCC cell line xenografts and human donor T cells in immunodeficient mice were used to support this study.Results Compared with a first-generation BTE, we show that the PBTE reduced avidity, intercellular synaptic strength, cytotoxic potency by as much as 33-fold, and ultimately efficacy against ccRCC tumors in vivo. However, compared with the PBTE, we demonstrate that the PMTE improved cell avidity, restored intercellular synapses, augmented cytotoxic potency by 40-fold, improved tumor distribution pharmacokinetics by 2-fold, and recovered synDNA efficacy in mouse tumor models by 20-fold. All the while, the PMTE displayed a desirable half-life of 4 days in mice compared with the conventional BTE’s 2 hours.Conclusions With impressive efficacy, the CA9-targeted PMTE is a promising new therapy for advanced ccRCC, which can be effectively delivered through synDNA. The highly potent PMTE format itself is a promising new tool for future applications in the multispecific antibody space.https://jitc.bmj.com/content/12/6/e008733.full
spellingShingle Carl H June
David Weiner
Christopher A Chuckran
Daniel Park
Ryan P O’Connell
Kevin Liaw
Nils Wellhausen
Pratik S Bhojnagarwala
Devivasha Bordoloi
Nicholas Shupin
Daniel Kulp
Format-tuning of in vivo-launched bispecific T cell engager enhances efficacy against renal cell carcinoma
Journal for ImmunoTherapy of Cancer
title Format-tuning of in vivo-launched bispecific T cell engager enhances efficacy against renal cell carcinoma
title_full Format-tuning of in vivo-launched bispecific T cell engager enhances efficacy against renal cell carcinoma
title_fullStr Format-tuning of in vivo-launched bispecific T cell engager enhances efficacy against renal cell carcinoma
title_full_unstemmed Format-tuning of in vivo-launched bispecific T cell engager enhances efficacy against renal cell carcinoma
title_short Format-tuning of in vivo-launched bispecific T cell engager enhances efficacy against renal cell carcinoma
title_sort format tuning of in vivo launched bispecific t cell engager enhances efficacy against renal cell carcinoma
url https://jitc.bmj.com/content/12/6/e008733.full
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