Deciphering molecular and cellular ex vivo responses to bispecific antibodies PD1-TIM3 and PD1-LAG3 in human tumors

Background Next-generation cancer immunotherapies are designed to broaden the therapeutic repertoire by targeting new immune checkpoints including lymphocyte-activation gene 3 (LAG-3) and T cell immunoglobulin and mucin-domain containing-3 (TIM-3). Yet, the molecular and cellular mechanisms by which...

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Main Authors: Alfred Zippelius, Petra Herzig, Pratiksha Gulati, Christian Klein, Marta Trüb, Kirsten D Mertz, Robert Rosenberg, Viola Heinzelmann-Schwarz, Mark Wiese, Didier Lardinois, Pablo Umana, Marina Natoli, Klas Hatje, Fabian Junker, Zhiwen Jiang, Iakov I Davydov, Markus Germann, Daniel Marbach, Adrian Zwick, Patrick Weber, Stefan Seeber, Lothar Tietze, Laura Codarri-Deak, Henry Kao
Format: Article
Language:English
Published: BMJ Publishing Group 2022-11-01
Series:Journal for ImmunoTherapy of Cancer
Online Access:https://jitc.bmj.com/content/10/11/e005548.full
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author Alfred Zippelius
Petra Herzig
Pratiksha Gulati
Christian Klein
Marta Trüb
Kirsten D Mertz
Robert Rosenberg
Viola Heinzelmann-Schwarz
Mark Wiese
Didier Lardinois
Pablo Umana
Marina Natoli
Klas Hatje
Fabian Junker
Zhiwen Jiang
Iakov I Davydov
Markus Germann
Daniel Marbach
Adrian Zwick
Patrick Weber
Stefan Seeber
Lothar Tietze
Laura Codarri-Deak
Henry Kao
author_facet Alfred Zippelius
Petra Herzig
Pratiksha Gulati
Christian Klein
Marta Trüb
Kirsten D Mertz
Robert Rosenberg
Viola Heinzelmann-Schwarz
Mark Wiese
Didier Lardinois
Pablo Umana
Marina Natoli
Klas Hatje
Fabian Junker
Zhiwen Jiang
Iakov I Davydov
Markus Germann
Daniel Marbach
Adrian Zwick
Patrick Weber
Stefan Seeber
Lothar Tietze
Laura Codarri-Deak
Henry Kao
author_sort Alfred Zippelius
collection DOAJ
description Background Next-generation cancer immunotherapies are designed to broaden the therapeutic repertoire by targeting new immune checkpoints including lymphocyte-activation gene 3 (LAG-3) and T cell immunoglobulin and mucin-domain containing-3 (TIM-3). Yet, the molecular and cellular mechanisms by which either receptor functions to mediate its inhibitory effects are still poorly understood. Similarly, little is known on the differential effects of dual, compared with single, checkpoint inhibition.Methods We here performed in-depth characterization, including multicolor flow cytometry, single cell RNA sequencing and multiplex supernatant analysis, using tumor single cell suspensions from patients with cancer treated ex vivo with novel bispecific antibodies targeting programmed cell death protein 1 (PD-1) and TIM-3 (PD1-TIM3), PD-1 and LAG-3 (PD1-LAG3), or with anti-PD-1.Results We identified patient samples which were responsive to PD1-TIM3, PD1-LAG3 or anti-PD-1 using an in vitro approach, validated by the analysis of 659 soluble proteins and enrichment for an anti-PD-1 responder signature. We found increased abundance of an activated (HLA-DR+CD25+GranzymeB+) CD8+ T cell subset and of proliferating CD8+ T cells, in response to bispecific antibody or anti-PD-1 treatment. Bispecific antibodies, but not anti-PD-1, significantly increased the abundance of a proliferating natural killer cell subset, which exhibited enrichment for a tissue-residency signature. Key phenotypic and transcriptional changes occurred in a PD-1+CXCL13+CD4+ T cell subset, in response to all treatments, including increased interleukin-17 secretion and signaling toward plasma cells. Interestingly, LAG-3 protein upregulation was detected as a unique pharmacodynamic effect mediated by PD1-LAG3, but not by PD1-TIM3 or anti-PD-1.Conclusions Our in vitro system reliably assessed responses to bispecific antibodies co-targeting PD-1 together with LAG-3 or TIM-3 using patients’ tumor infiltrating immune cells and revealed transcriptional and phenotypic imprinting by bispecific antibody formats currently tested in early clinical trials.
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series Journal for ImmunoTherapy of Cancer
spelling doaj-art-d05c988c40044c0488968cfb22947d692025-02-09T12:15:15ZengBMJ Publishing GroupJournal for ImmunoTherapy of Cancer2051-14262022-11-01101110.1136/jitc-2022-005548Deciphering molecular and cellular ex vivo responses to bispecific antibodies PD1-TIM3 and PD1-LAG3 in human tumorsAlfred Zippelius0Petra Herzig1Pratiksha Gulati2Christian Klein3Marta Trüb4Kirsten D Mertz5Robert Rosenberg6Viola Heinzelmann-Schwarz7Mark Wiese8Didier Lardinois9Pablo Umana10Marina Natoli11Klas Hatje12Fabian Junker13Zhiwen Jiang14Iakov I Davydov15Markus Germann16Daniel Marbach17Adrian Zwick18Patrick Weber19Stefan Seeber20Lothar Tietze21Laura Codarri-Deak22Henry Kao239 Medical Oncology, University Hospital Basel, Basel, Switzerland1 Department of Biomedicine, University Hospital Basel, Basel, SwitzerlandRoche Pharma Research and Early Development, Pharmaceutical Sciences, Roche Innovation Center Basel, F Hoffmann-La Roche Ltd, Basel, SwitzerlandRoche Glycart AG, Schlieren, SwitzerlandDepartment of Biomedicine, University Hospital Basel, Basel, Switzerland8 Institute of Pathology, Cantonal Hospital Basel-Landschaft, Liestal, Switzerland8 Department of Surgery, Cantonal Hospital Basel-Landschaft, Liestal, Switzerland7 Department of Gynecology and Obstetrics, University Hospital Basel, Basel, Switzerland10 Division of Thoracic Surgery, University Hospital Basel, Basel, Switzerland6 Department of Thoracic Surgery, University Hospital Basel, Basel, Switzerland6Roche Pharmaceutical Research and Early Development (pRED), Roche Innovation Center Zurich (RICZ), Schlieren, Switzerland1 Department of Biomedicine, University Hospital Basel, Basel, SwitzerlandRoche Pharma Research and Early Development, Pharmaceutical Sciences, Roche Innovation Center Basel, F Hoffmann-La Roche Ltd, Basel, SwitzerlandRoche Pharma Research and Early Development, Pharmaceutical Sciences, Roche Innovation Center Basel, F Hoffmann-La Roche Ltd, Basel, Switzerland5Roche Innovation Center Basel, Roche Pharma Research and Early Development, Basel, SwitzerlandRoche Pharma Research and Early Development, Pharmaceutical Sciences, Roche Innovation Center, F. Hoffmann-La Roche Ltd, Basel, SwitzerlandDepartment of Biomedicine, University Hospital Basel, Basel, SwitzerlandRoche Pharma Research and Early Development, Pharmaceutical Sciences, Roche Innovation Center Basel, F Hoffmann-La Roche Ltd, Basel, SwitzerlandRoche Pharma Research and Early Development, Pharmaceutical Sciences, Roche Innovation Center Munich, F Hoffmann-La Roche Ltd, Penzberg, GermanyRoche Pharma Research and Early Development, Discovery Oncology, Roche Innovation Center Zurich, Schlieren, SwitzerlandRoche Pharma Research and Early Development, Pharmaceutical Sciences, Roche Innovation Center Munich, F Hoffmann-La Roche Ltd, Penzberg, GermanyInstitute of Pathology, Lahr, GermanyRoche Pharma Research and Early Development, Discovery Oncology, Roche Innovation Center Zurich, Schlieren, SwitzerlandRoche Pharma Research and Early Development, Early Biomarker Development Oncology, Roche Innovation Center Basel, F Hoffmann-La Roche Ltd, Basel, SwitzerlandBackground Next-generation cancer immunotherapies are designed to broaden the therapeutic repertoire by targeting new immune checkpoints including lymphocyte-activation gene 3 (LAG-3) and T cell immunoglobulin and mucin-domain containing-3 (TIM-3). Yet, the molecular and cellular mechanisms by which either receptor functions to mediate its inhibitory effects are still poorly understood. Similarly, little is known on the differential effects of dual, compared with single, checkpoint inhibition.Methods We here performed in-depth characterization, including multicolor flow cytometry, single cell RNA sequencing and multiplex supernatant analysis, using tumor single cell suspensions from patients with cancer treated ex vivo with novel bispecific antibodies targeting programmed cell death protein 1 (PD-1) and TIM-3 (PD1-TIM3), PD-1 and LAG-3 (PD1-LAG3), or with anti-PD-1.Results We identified patient samples which were responsive to PD1-TIM3, PD1-LAG3 or anti-PD-1 using an in vitro approach, validated by the analysis of 659 soluble proteins and enrichment for an anti-PD-1 responder signature. We found increased abundance of an activated (HLA-DR+CD25+GranzymeB+) CD8+ T cell subset and of proliferating CD8+ T cells, in response to bispecific antibody or anti-PD-1 treatment. Bispecific antibodies, but not anti-PD-1, significantly increased the abundance of a proliferating natural killer cell subset, which exhibited enrichment for a tissue-residency signature. Key phenotypic and transcriptional changes occurred in a PD-1+CXCL13+CD4+ T cell subset, in response to all treatments, including increased interleukin-17 secretion and signaling toward plasma cells. Interestingly, LAG-3 protein upregulation was detected as a unique pharmacodynamic effect mediated by PD1-LAG3, but not by PD1-TIM3 or anti-PD-1.Conclusions Our in vitro system reliably assessed responses to bispecific antibodies co-targeting PD-1 together with LAG-3 or TIM-3 using patients’ tumor infiltrating immune cells and revealed transcriptional and phenotypic imprinting by bispecific antibody formats currently tested in early clinical trials.https://jitc.bmj.com/content/10/11/e005548.full
spellingShingle Alfred Zippelius
Petra Herzig
Pratiksha Gulati
Christian Klein
Marta Trüb
Kirsten D Mertz
Robert Rosenberg
Viola Heinzelmann-Schwarz
Mark Wiese
Didier Lardinois
Pablo Umana
Marina Natoli
Klas Hatje
Fabian Junker
Zhiwen Jiang
Iakov I Davydov
Markus Germann
Daniel Marbach
Adrian Zwick
Patrick Weber
Stefan Seeber
Lothar Tietze
Laura Codarri-Deak
Henry Kao
Deciphering molecular and cellular ex vivo responses to bispecific antibodies PD1-TIM3 and PD1-LAG3 in human tumors
Journal for ImmunoTherapy of Cancer
title Deciphering molecular and cellular ex vivo responses to bispecific antibodies PD1-TIM3 and PD1-LAG3 in human tumors
title_full Deciphering molecular and cellular ex vivo responses to bispecific antibodies PD1-TIM3 and PD1-LAG3 in human tumors
title_fullStr Deciphering molecular and cellular ex vivo responses to bispecific antibodies PD1-TIM3 and PD1-LAG3 in human tumors
title_full_unstemmed Deciphering molecular and cellular ex vivo responses to bispecific antibodies PD1-TIM3 and PD1-LAG3 in human tumors
title_short Deciphering molecular and cellular ex vivo responses to bispecific antibodies PD1-TIM3 and PD1-LAG3 in human tumors
title_sort deciphering molecular and cellular ex vivo responses to bispecific antibodies pd1 tim3 and pd1 lag3 in human tumors
url https://jitc.bmj.com/content/10/11/e005548.full
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