Novel cytokine–antibody fusion protein, N-820, to enhance the functions of ex vivo expanded natural killer cells against Burkitt lymphoma

Background The prognosis of patients with relapsed or progressive B cell (CD20+) non-Hodgkin’s lymphoma (B-NHL), including Burkitt lymphoma (BL), is dismal due to chemoradiotherapy resistance. Novel therapeutic strategies are urgently needed. N-820 is a fusion protein of N-803 (formerly known as ALT...

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Main Authors: John Lee, Dean Lee, Patrick Soon-Shiong, Yaya Chu, Gaurav Nayyar, Nang Kham Su, Jeremy M Rosenblum, Jeffrey T Safrit, Matthew Barth, Mitchell S Cairo
Format: Article
Language:English
Published: BMJ Publishing Group 2020-10-01
Series:Journal for ImmunoTherapy of Cancer
Online Access:https://jitc.bmj.com/content/8/2/e001238.full
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author John Lee
Dean Lee
Patrick Soon-Shiong
Yaya Chu
Gaurav Nayyar
Nang Kham Su
Jeremy M Rosenblum
Jeffrey T Safrit
Matthew Barth
Mitchell S Cairo
author_facet John Lee
Dean Lee
Patrick Soon-Shiong
Yaya Chu
Gaurav Nayyar
Nang Kham Su
Jeremy M Rosenblum
Jeffrey T Safrit
Matthew Barth
Mitchell S Cairo
author_sort John Lee
collection DOAJ
description Background The prognosis of patients with relapsed or progressive B cell (CD20+) non-Hodgkin’s lymphoma (B-NHL), including Burkitt lymphoma (BL), is dismal due to chemoradiotherapy resistance. Novel therapeutic strategies are urgently needed. N-820 is a fusion protein of N-803 (formerly known as ALT-803) to four single-chains of rituximab. This agent has tri-specific binding activity to CD20 and enhanced antibody-dependent cell-mediated cytotoxicity.Methods We investigated the anti-tumor combinatorial effects of N-820 with ex vivo expanded peripheral blood natural killer (exPBNK) cells against rituximab-sensitive and rituximab-resistant CD20+ BL in vitro using cytoxicity assays and in vivo using human BL xenografted NOD/SCID/IL2rγnull (NSG) mice. We also investigated the cytokines/chemokines/growth factors released using ELISA and multiplex assay. Gene expression changes were examined using real-time PCR arrays.Results N-820 significantly enhanced the expression of NK activating receptors (p<0.001) and the proliferation of exPBNK cells with enhanced Ki67 expression and Stat5 phosphorylation (p<0.001). N-820 significantly enhanced the secretion of cytokines, chemokines, and growth factors including GM-CSF, RANTES, MIP-1B (p<0.001) from exPBNK cells as compared with the combination of rituximab+N-803. Importantly, N-820 significantly enhanced in vitro cytotoxicity (p<0.001) of exPBNK with enhanced granzyme B and IFN-γ release (p<0.001) against BL. Gene expression profiles in exPBNK stimulated by N-820+Raji-2R showed enhanced transcription of CXCL9, CXCL1, CSF2, CSF3, GZMB, and IFNG. Moreover, N-820 combined with exPBNK significantly inhibited rituximab-resistant BL growth (p<0.05) and extended the survival (p<0.05) of BL xenografted NSG mice.Conclusions Our results provide the rationale for the development of a clinical trial of N-820 alone or in combination with endogenous or ex vivo expanded NK cells in patients with CD20+ B-NHL failing prior rituximab containing chemoimmunotherapy regimens.
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series Journal for ImmunoTherapy of Cancer
spelling doaj-art-752d96bf91744f83a32d2522239e2b102024-11-10T18:30:08ZengBMJ Publishing GroupJournal for ImmunoTherapy of Cancer2051-14262020-10-018210.1136/jitc-2020-001238Novel cytokine–antibody fusion protein, N-820, to enhance the functions of ex vivo expanded natural killer cells against Burkitt lymphomaJohn Lee0Dean Lee1Patrick Soon-Shiong2Yaya Chu3Gaurav Nayyar4Nang Kham Su5Jeremy M Rosenblum6Jeffrey T Safrit7Matthew Barth8Mitchell S Cairo9Hepatology, Galway University Hospitals, Galway, IrelandPediatric Hem/Onc/BMT, Nationwide Children`s Hospital Hematology Oncology and Blood and Marrow Transplant, Columbus, Ohio, USAImmunityBio, Inc, Culver City, California, USADepartment of Pediatrics, New York Medical College, Valhalla, New York, USADepartment of Pediatrics, New York Medical College, Valhalla, New York, USA1 Department of Pediatrics, New York Medical College, Valhalla, New York, USADepartment of Pediatrics, New York Medical College, Valhalla, New York, USANantKwest, Culver City, California, USA4 Department of Pediatrics, State University of New York at Buffalo, Buffalo, New York, USA9 Department of Cell Biology and Anatomy, New York Medical College, Valhalla, New York, USABackground The prognosis of patients with relapsed or progressive B cell (CD20+) non-Hodgkin’s lymphoma (B-NHL), including Burkitt lymphoma (BL), is dismal due to chemoradiotherapy resistance. Novel therapeutic strategies are urgently needed. N-820 is a fusion protein of N-803 (formerly known as ALT-803) to four single-chains of rituximab. This agent has tri-specific binding activity to CD20 and enhanced antibody-dependent cell-mediated cytotoxicity.Methods We investigated the anti-tumor combinatorial effects of N-820 with ex vivo expanded peripheral blood natural killer (exPBNK) cells against rituximab-sensitive and rituximab-resistant CD20+ BL in vitro using cytoxicity assays and in vivo using human BL xenografted NOD/SCID/IL2rγnull (NSG) mice. We also investigated the cytokines/chemokines/growth factors released using ELISA and multiplex assay. Gene expression changes were examined using real-time PCR arrays.Results N-820 significantly enhanced the expression of NK activating receptors (p<0.001) and the proliferation of exPBNK cells with enhanced Ki67 expression and Stat5 phosphorylation (p<0.001). N-820 significantly enhanced the secretion of cytokines, chemokines, and growth factors including GM-CSF, RANTES, MIP-1B (p<0.001) from exPBNK cells as compared with the combination of rituximab+N-803. Importantly, N-820 significantly enhanced in vitro cytotoxicity (p<0.001) of exPBNK with enhanced granzyme B and IFN-γ release (p<0.001) against BL. Gene expression profiles in exPBNK stimulated by N-820+Raji-2R showed enhanced transcription of CXCL9, CXCL1, CSF2, CSF3, GZMB, and IFNG. Moreover, N-820 combined with exPBNK significantly inhibited rituximab-resistant BL growth (p<0.05) and extended the survival (p<0.05) of BL xenografted NSG mice.Conclusions Our results provide the rationale for the development of a clinical trial of N-820 alone or in combination with endogenous or ex vivo expanded NK cells in patients with CD20+ B-NHL failing prior rituximab containing chemoimmunotherapy regimens.https://jitc.bmj.com/content/8/2/e001238.full
spellingShingle John Lee
Dean Lee
Patrick Soon-Shiong
Yaya Chu
Gaurav Nayyar
Nang Kham Su
Jeremy M Rosenblum
Jeffrey T Safrit
Matthew Barth
Mitchell S Cairo
Novel cytokine–antibody fusion protein, N-820, to enhance the functions of ex vivo expanded natural killer cells against Burkitt lymphoma
Journal for ImmunoTherapy of Cancer
title Novel cytokine–antibody fusion protein, N-820, to enhance the functions of ex vivo expanded natural killer cells against Burkitt lymphoma
title_full Novel cytokine–antibody fusion protein, N-820, to enhance the functions of ex vivo expanded natural killer cells against Burkitt lymphoma
title_fullStr Novel cytokine–antibody fusion protein, N-820, to enhance the functions of ex vivo expanded natural killer cells against Burkitt lymphoma
title_full_unstemmed Novel cytokine–antibody fusion protein, N-820, to enhance the functions of ex vivo expanded natural killer cells against Burkitt lymphoma
title_short Novel cytokine–antibody fusion protein, N-820, to enhance the functions of ex vivo expanded natural killer cells against Burkitt lymphoma
title_sort novel cytokine antibody fusion protein n 820 to enhance the functions of ex vivo expanded natural killer cells against burkitt lymphoma
url https://jitc.bmj.com/content/8/2/e001238.full
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