Engineered IL-18 variants with half-life extension and improved stability for cancer immunotherapy

Background The pro-inflammatory cytokine, interleukin-18 (IL-18), plays an instrumental role in bolstering anti-tumor immunity. However, the therapeutic application of IL-18 has been limited due to its susceptibility to neutralization by IL-18 binding protein (IL-18BP), short in vivo half-life, and...

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Main Authors: Yue Zhao, Yan Qu, Marina Moskalenko, Dawei Sun, Travis W Bainbridge, Matthieu Masureel, Lifen Wang, Robert Herrera, Maciej T Paluch, Kazi Tasneem, Harpinder Saini, Manal Sadek, Mandy Kwong, Yoon Min Kim, Jay M Bhatt, Christine Tam, Pamela Pui Fung Chan, Ayse Meric Ovacik, Jonathan T Sockolosky, Nathaniel R West, Beyza Bulutoglu
Format: Article
Language:English
Published: BMJ Publishing Group 2025-07-01
Series:Journal for ImmunoTherapy of Cancer
Online Access:https://jitc.bmj.com/content/13/7/e011789.full
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author Yue Zhao
Yan Qu
Marina Moskalenko
Dawei Sun
Travis W Bainbridge
Matthieu Masureel
Lifen Wang
Robert Herrera
Maciej T Paluch
Kazi Tasneem
Harpinder Saini
Manal Sadek
Mandy Kwong
Yoon Min Kim
Jay M Bhatt
Christine Tam
Pamela Pui Fung Chan
Ayse Meric Ovacik
Jonathan T Sockolosky
Nathaniel R West
Beyza Bulutoglu
author_facet Yue Zhao
Yan Qu
Marina Moskalenko
Dawei Sun
Travis W Bainbridge
Matthieu Masureel
Lifen Wang
Robert Herrera
Maciej T Paluch
Kazi Tasneem
Harpinder Saini
Manal Sadek
Mandy Kwong
Yoon Min Kim
Jay M Bhatt
Christine Tam
Pamela Pui Fung Chan
Ayse Meric Ovacik
Jonathan T Sockolosky
Nathaniel R West
Beyza Bulutoglu
author_sort Yue Zhao
collection DOAJ
description Background The pro-inflammatory cytokine, interleukin-18 (IL-18), plays an instrumental role in bolstering anti-tumor immunity. However, the therapeutic application of IL-18 has been limited due to its susceptibility to neutralization by IL-18 binding protein (IL-18BP), short in vivo half-life, and unfavorable physicochemical properties.Methods In order to overcome the poor drug-like properties of IL-18, we installed an artificial disulfide bond, removed the native, unpaired cysteines, and fused the stabilized cytokine to an IgG Fc domain. The stability, potency, pharmacokinetic and pharmacodynamic properties as well as efficacy of disulfide-stabilized IL-18 Fc-fusion (dsIL-18-Fc) were assessed via in vitro and in vivo studies.Results The stability and mammalian host cell production yields of dsIL-18-Fc were improved, compared to the wild-type (WT) cytokine, while maintaining its biological potency and interactions with IL-18 receptor α (IL-18Rα) and IL-18BP. Recombinant fusion of the cytokine to an IgG Fc domain provided extended half-life. Notably, despite maintaining sensitivity to IL-18BP, dsIL-18-Fc was effective at activating both T and natural killer (NK) cells, and elicited a strong anti-tumor response, either as a single agent, or in conjunction with anti-programmed cell death-ligand 1 (anti-PD-L1) therapy.Conclusions We engineered IL-18 for reinforced stability, extended half-life, and improved manufacturability. The therapeutic benefit of dsIL-18-Fc, coupled with a more favorable manufacturability profile and enhanced drug-like properties, underscores the potential utility of this engineered cytokine in cancer immunotherapy.
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spelling doaj-art-0eaa0c2f92024555b88b4cb2a4bbaaaa2025-08-20T03:27:48ZengBMJ Publishing GroupJournal for ImmunoTherapy of Cancer2051-14262025-07-0113710.1136/jitc-2025-011789Engineered IL-18 variants with half-life extension and improved stability for cancer immunotherapyYue Zhao0Yan Qu1Marina Moskalenko2Dawei Sun3Travis W Bainbridge4Matthieu Masureel5Lifen Wang6Robert Herrera7Maciej T Paluch8Kazi Tasneem9Harpinder Saini10Manal Sadek11Mandy Kwong12Yoon Min Kim13Jay M Bhatt14Christine Tam15Pamela Pui Fung Chan16Ayse Meric Ovacik17Jonathan T Sockolosky18Nathaniel R West19Beyza Bulutoglu204 Biochemical and Cellular Pharmacology, Genentech Inc, South San Francisco, California, USA3 Translational Oncology, Genentech Inc, South San Francisco, California, USA3 Translational Oncology, Genentech Inc, South San Francisco, California, USA1 Protein Sciences, Genentech Inc, South San Francisco, California, USA1 Protein Sciences, Genentech Inc, South San Francisco, California, USA1 Protein Sciences, Genentech Inc, South San Francisco, California, USA2 Discovery Oncology, Genentech Inc, South San Francisco, California, USA4 Biochemical and Cellular Pharmacology, Genentech Inc, South San Francisco, California, USA1 Protein Sciences, Genentech Inc, South San Francisco, California, USA5 Translational Pharmacokinetics and Pharmacodynamics, Genentech Inc, South San Francisco, California, USA4 Biochemical and Cellular Pharmacology, Genentech Inc, South San Francisco, California, USA4 Biochemical and Cellular Pharmacology, Genentech Inc, South San Francisco, California, USA4 Biochemical and Cellular Pharmacology, Genentech Inc, South San Francisco, California, USA1 Protein Sciences, Genentech Inc, South San Francisco, California, USA1 Protein Sciences, Genentech Inc, South San Francisco, California, USA1 Protein Sciences, Genentech Inc, South San Francisco, California, USA4 Biochemical and Cellular Pharmacology, Genentech Inc, South San Francisco, California, USA5 Translational Pharmacokinetics and Pharmacodynamics, Genentech Inc, South San Francisco, California, USA6 Antibody Engineering, Genentech Inc, South San Francisco, California, USA2 Discovery Oncology, Genentech Inc, South San Francisco, California, USA1 Protein Sciences, Genentech Inc, South San Francisco, California, USABackground The pro-inflammatory cytokine, interleukin-18 (IL-18), plays an instrumental role in bolstering anti-tumor immunity. However, the therapeutic application of IL-18 has been limited due to its susceptibility to neutralization by IL-18 binding protein (IL-18BP), short in vivo half-life, and unfavorable physicochemical properties.Methods In order to overcome the poor drug-like properties of IL-18, we installed an artificial disulfide bond, removed the native, unpaired cysteines, and fused the stabilized cytokine to an IgG Fc domain. The stability, potency, pharmacokinetic and pharmacodynamic properties as well as efficacy of disulfide-stabilized IL-18 Fc-fusion (dsIL-18-Fc) were assessed via in vitro and in vivo studies.Results The stability and mammalian host cell production yields of dsIL-18-Fc were improved, compared to the wild-type (WT) cytokine, while maintaining its biological potency and interactions with IL-18 receptor α (IL-18Rα) and IL-18BP. Recombinant fusion of the cytokine to an IgG Fc domain provided extended half-life. Notably, despite maintaining sensitivity to IL-18BP, dsIL-18-Fc was effective at activating both T and natural killer (NK) cells, and elicited a strong anti-tumor response, either as a single agent, or in conjunction with anti-programmed cell death-ligand 1 (anti-PD-L1) therapy.Conclusions We engineered IL-18 for reinforced stability, extended half-life, and improved manufacturability. The therapeutic benefit of dsIL-18-Fc, coupled with a more favorable manufacturability profile and enhanced drug-like properties, underscores the potential utility of this engineered cytokine in cancer immunotherapy.https://jitc.bmj.com/content/13/7/e011789.full
spellingShingle Yue Zhao
Yan Qu
Marina Moskalenko
Dawei Sun
Travis W Bainbridge
Matthieu Masureel
Lifen Wang
Robert Herrera
Maciej T Paluch
Kazi Tasneem
Harpinder Saini
Manal Sadek
Mandy Kwong
Yoon Min Kim
Jay M Bhatt
Christine Tam
Pamela Pui Fung Chan
Ayse Meric Ovacik
Jonathan T Sockolosky
Nathaniel R West
Beyza Bulutoglu
Engineered IL-18 variants with half-life extension and improved stability for cancer immunotherapy
Journal for ImmunoTherapy of Cancer
title Engineered IL-18 variants with half-life extension and improved stability for cancer immunotherapy
title_full Engineered IL-18 variants with half-life extension and improved stability for cancer immunotherapy
title_fullStr Engineered IL-18 variants with half-life extension and improved stability for cancer immunotherapy
title_full_unstemmed Engineered IL-18 variants with half-life extension and improved stability for cancer immunotherapy
title_short Engineered IL-18 variants with half-life extension and improved stability for cancer immunotherapy
title_sort engineered il 18 variants with half life extension and improved stability for cancer immunotherapy
url https://jitc.bmj.com/content/13/7/e011789.full
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