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Rituximab and obinutuzumab differentially hijack the B-cell receptor and NOTCH1 signaling pathways

Rituximab and obinutuzumab differentially hijack the B-cell receptor and NOTCH1 signaling pathways
Rituximab and obinutuzumab differentially hijack the B-cell receptor and NOTCH1 signaling pathways
The anti-CD20 monoclonal antibodies rituximab and obinutuzumab differ in their mechanisms of action, with obinutuzumab evoking greater direct B cell death. To characterize the signaling processes responsible for improved B cell killing by obinutuzumab, we undertook a phosphoproteomics approach and demonstrate that rituximab and obinutuzumab differentially activate pathways downstream of the B cell receptor. Although both antibodies induce strong ERK and MYC activation sufficient to promote cell-cycle arrest and B cell death, obinutuzumab exceeds rituximab in supporting apoptosis induction by means of aberrant SYK phosphorylation. In contrast, rituximab elicits stronger anti-apoptotic signals by activating AKT, by impairing pro-apoptotic BAD, and by releasing membrane-bound NOTCH1 to up-regulate pro-survival target genes. As a consequence, rituximab appears to reinforce BCL2-mediated apoptosis resistance. The unexpected complexity and differences by which rituximab and obinutuzumab interfere with signaling pathways essential for lymphoma pathogenesis and treatment provide important impetus to optimize and personalize the application of different anti-CD20 treatments.
cancer, immunology, systems biology: proteomics
Edelmann, Jennifer
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Dokal, Arran D.
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Vilventhraraja, Emma
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Holzmann, Karlheinz
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Britton, David
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Klymenko, Tetyana
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Döhner, Hartmut
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Cragg, Mark
ec97f80e-f3c8-49b7-a960-20dff648b78c
Braun, Andrejs
a40f851b-1cd7-4ce7-8c25-832e38950f52
Cutillas, Pedro
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Gribben, John G.
086dd67d-a2d9-4d2c-97d6-8e345eb84142
Edelmann, Jennifer
573e8a7e-4e3b-4007-b2fd-3a136cbe5baa
Dokal, Arran D.
bdf951d9-133a-413d-9c6e-10d7093eb011
Vilventhraraja, Emma
f794d50c-bd2c-4f18-8a2f-302eaf557ff2
Holzmann, Karlheinz
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Britton, David
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Klymenko, Tetyana
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Döhner, Hartmut
6e7be004-744b-484d-ae35-32f5941c3b96
Cragg, Mark
ec97f80e-f3c8-49b7-a960-20dff648b78c
Braun, Andrejs
a40f851b-1cd7-4ce7-8c25-832e38950f52
Cutillas, Pedro
e097269e-e672-4e73-a11c-06d162b5fa24
Gribben, John G.
086dd67d-a2d9-4d2c-97d6-8e345eb84142

Edelmann, Jennifer, Dokal, Arran D., Vilventhraraja, Emma, Holzmann, Karlheinz, Britton, David, Klymenko, Tetyana, Döhner, Hartmut, Cragg, Mark, Braun, Andrejs, Cutillas, Pedro and Gribben, John G. (2021) Rituximab and obinutuzumab differentially hijack the B-cell receptor and NOTCH1 signaling pathways. iScience, 24 (2), [102089]. (doi:10.1016/j.isci.2021.102089).

Record type: Article

Abstract

The anti-CD20 monoclonal antibodies rituximab and obinutuzumab differ in their mechanisms of action, with obinutuzumab evoking greater direct B cell death. To characterize the signaling processes responsible for improved B cell killing by obinutuzumab, we undertook a phosphoproteomics approach and demonstrate that rituximab and obinutuzumab differentially activate pathways downstream of the B cell receptor. Although both antibodies induce strong ERK and MYC activation sufficient to promote cell-cycle arrest and B cell death, obinutuzumab exceeds rituximab in supporting apoptosis induction by means of aberrant SYK phosphorylation. In contrast, rituximab elicits stronger anti-apoptotic signals by activating AKT, by impairing pro-apoptotic BAD, and by releasing membrane-bound NOTCH1 to up-regulate pro-survival target genes. As a consequence, rituximab appears to reinforce BCL2-mediated apoptosis resistance. The unexpected complexity and differences by which rituximab and obinutuzumab interfere with signaling pathways essential for lymphoma pathogenesis and treatment provide important impetus to optimize and personalize the application of different anti-CD20 treatments.

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Accepted/In Press date: 18 January 2021
e-pub ahead of print date: 22 January 2021
Published date: 19 February 2021
Additional Information: Funding Information: The study was supported by the German Research Foundation ( ED 256/1-1 ), the Barts Charity Fund, NIH ( NCI PO1 CA81534 ), the Barry Reed Cancer Research Fund, and the Clinician Scientist Program of the Medical Faculty, Ulm University . Funding Information: We thank Sameena Iqubal and Janet Matthews from Barts Cancer Institute for their help with the acquisition of primary CLL samples, Ryan Smith from Barts Cancer Institute for his help with the analysis of LC-MS/MS raw data, Graham Packham and Stephen Beers from Southampton University for helpful discussions, and Doriana di Bella from Barts Cancer Institute for her assistance with experiments. The study was supported by the German Research Foundation (ED 256/1-1), the Barts Charity Fund, NIH (NCI PO1 CA81534), the Barry Reed Cancer Research Fund, and the Clinician Scientist Program of the Medical Faculty, Ulm University. All authors contributed extensively to the work presented in this paper. J.E. designed research; performed, analyzed, and interpreted experiments; and wrote the paper. A.D.D. designed and performed the LC-MS/MS experiment, analyzed the raw data, and contributed to interpretation of the LC-MS/MS experiment. E.V. designed and performed the LC-MS/MS experiment. K.H. analyzed experimental data. M.C. and A.B. contributed to research design and data interpretation. H.D. contributed to data interpretation. P.C. contributed to design, analysis, and interpretation of the LC-MS/MS experiment. J.G.G. contributed to research design and interpretation. All authors contributed to write the paper. A.D.D. is Senior Scientist at Kinomica Ltd. E.V. is employed by The Janssen Pharmaceutical Companies of Johnson & Johnson. D.B. is Academic Funder and CSO at Kinomica Ltd. M.C. acts as a consultant for a number of biotech companies, being retained as a consultant for BioInvent and has received research funding from BioInvent, GSK, UCB, iTeos, and Roche. P.C. is Academic Funder and Director at Kinomica Ltd. Disclosures J.G.G.: Janssen: Advisory Board, Honoraria, Research funding; Acerta: Advisory Board, Honoraria, Research funding; Celgene: Advisory Board, Honoraria, Research funding; Kite: Advisory Board, Honoraria; AbbVie: Advisory Board, Honoraria; Novartis: Advisory Board; TG Therapeutics: Advisory Board. Disclosures H.D.: Abbvie: Consultancy, Honoraria; Agios: Consultancy, Honoraria, Research funding; Amgen: Consultancy, Honoraria, Research Funding; AROG: Research funding; Astellas: Consultancy, Honoraria, Research funding; Astex: Consultancy, Honoraria; Bristol Myers Squibb: Consultancy, Honoraria, Research Funding; Celgene: Consultancy, Honoraria, Research funding; Janssen: Consultancy, Honoraria; Jazz: Consultancy, Honoraria, Research Funding; Helsinn: Consultancy, Honoraria; Novartis: Consultancy, Honoraria, Research funding; Oxford Biomedicals: Consultancy, Honoraria; Pfizer: Research funding; Roche: Consultancy, Honoraria; Sunesis: Research funding; AstraZeneca: Consultancy, Honoraria; GEMoaB: Consultancy, Honoraria. The remaining authors declare no competing interests. Funding Information: M.C. acts as a consultant for a number of biotech companies, being retained as a consultant for BioInvent and has received research funding from BioInvent , GSK , UCB , iTeos , and Roche . Funding Information: Disclosures H.D.: Abbvie: Consultancy, Honoraria; Agios : Consultancy, Honoraria, Research funding; Amgen : Consultancy, Honoraria, Research Funding; AROG : Research funding; Astellas : Consultancy, Honoraria, Research funding; Astex: Consultancy, Honoraria; Bristol Myers Squibb : Consultancy, Honoraria, Research Funding; Celgene : Consultancy, Honoraria, Research funding; Janssen : Consultancy, Honoraria; Jazz: Consultancy, Honoraria, Research Funding; Helsinn : Consultancy, Honoraria; Novartis: Consultancy, Honoraria, Research funding; Oxford Biomedicals : Consultancy, Honoraria; Pfizer: Research funding; Roche : Consultancy, Honoraria; Sunesis: Research funding; AstraZeneca: Consultancy, Honoraria; GEMoaB: Consultancy, Honoraria. Publisher Copyright: © 2021 The Authors
Keywords: cancer, immunology, systems biology: proteomics

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Local EPrints ID: 446918
URI: http://eprints.soton.ac.uk/id/eprint/446918
PURE UUID: 6708cde8-5186-4ea9-99b8-22fa8cae4548
ORCID for Mark Cragg: ORCID iD orcid.org/0000-0003-2077-089X

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Date deposited: 26 Feb 2021 17:31
Last modified: 17 Mar 2024 02:46

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Contributors

Author: Jennifer Edelmann
Author: Arran D. Dokal
Author: Emma Vilventhraraja
Author: Karlheinz Holzmann
Author: David Britton
Author: Tetyana Klymenko
Author: Hartmut Döhner
Author: Mark Cragg ORCID iD
Author: Andrejs Braun
Author: Pedro Cutillas
Author: John G. Gribben

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