Novel trispecific killer engager targeting B7-H3 enhances natural killer cell antitumor activity against head and neck cancer

Background Patients with head and neck squamous cell carcinoma (HNSCC), particularly the human papillomavirus negative (HPV−) subset, have a dismal prognosis. Furthermore, patients with Fanconi anemia (FA) have a genetic predisposition with a 500-fold to 700-fold higher incidence of HNSCC. Thus, nov...

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Main Authors: Jeffrey S Miller, Martin Felices, Peter Hinderlie, James Lim, Naomi Fujioka, Shee Kwan Phung, Yvette Soignier, Joshua T Walker, Melissa A Geller, Madison Shackelford, Carly Selleck, Philippa R Kennedy, Mihir Shetty, Melissa J Khaw, Paolo P Provenzano, Nicholas A Zorko, Laura E Bendzick, Riley C Lyons, Elise L Femino, Terran Stenger, Tumpa Dasgupta, Laura E Kotz, Eng Hock Lee, Quynhanh Lu, Zachary B Davis, John E Wagner, Margaret L MacMillan
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/e011370.full
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author Jeffrey S Miller
Martin Felices
Peter Hinderlie
James Lim
Naomi Fujioka
Shee Kwan Phung
Yvette Soignier
Joshua T Walker
Melissa A Geller
Madison Shackelford
Carly Selleck
Philippa R Kennedy
Mihir Shetty
Melissa J Khaw
Paolo P Provenzano
Nicholas A Zorko
Laura E Bendzick
Riley C Lyons
Elise L Femino
Terran Stenger
Tumpa Dasgupta
Laura E Kotz
Eng Hock Lee
Quynhanh Lu
Zachary B Davis
John E Wagner
Margaret L MacMillan
author_facet Jeffrey S Miller
Martin Felices
Peter Hinderlie
James Lim
Naomi Fujioka
Shee Kwan Phung
Yvette Soignier
Joshua T Walker
Melissa A Geller
Madison Shackelford
Carly Selleck
Philippa R Kennedy
Mihir Shetty
Melissa J Khaw
Paolo P Provenzano
Nicholas A Zorko
Laura E Bendzick
Riley C Lyons
Elise L Femino
Terran Stenger
Tumpa Dasgupta
Laura E Kotz
Eng Hock Lee
Quynhanh Lu
Zachary B Davis
John E Wagner
Margaret L MacMillan
author_sort Jeffrey S Miller
collection DOAJ
description Background Patients with head and neck squamous cell carcinoma (HNSCC), particularly the human papillomavirus negative (HPV−) subset, have a dismal prognosis. Furthermore, patients with Fanconi anemia (FA) have a genetic predisposition with a 500-fold to 700-fold higher incidence of HNSCC. Thus, novel and more efficacious therapies are needed. As current immunotherapies often fail due to suppressive elements in the tumor microenvironment (TME), we developed a trispecific killer engager (TriKE) to direct multiple signals to natural killer (NK) cells to overcome the hypoxic TME. This TriKE is comprised of a camelid nanobody that binds to CD16 on NK cells, an interleukin (IL)-15 moiety, and another novel camelid nanobody that binds to the B7-H3 antigen, which is highly and specifically expressed on the tumor cell surface.Methods The B7H3 TriKE was generated using a mammalian expression system. Its functionality was evaluated using flow cytometry-based NK cell degranulation, cytokine production, proliferation and live cell imaging cytotoxicity assays. Models of acute and prolonged hypoxia (1% oxygen) were carried out to assess tumor killing. Tumor progression, NK cell persistence, and survival differences between IL-15-treated and TriKE-treated mice were studied using NOD-scidIL2Rgnull (NSG) mice engrafted with human HNSCC.Results High B7-H3 expression was found in HPV− HNSCC cell lines, even when the FA gene was knocked out, and The Cancer Genome Atlas patient data showed that high B7-H3 expression predicted poor survival in patients with HPV− HNSCC. Similar to the NK cell activity seen with healthy donors, the B7H3 TriKE enhanced activation, expansion and cytotoxicity of NK cells from patients with HPV− HNSCC, a target population for this therapeutic. Additionally, the B7H3 TriKE improved NK cell cytotoxicity in a three-dimensional spheroid model of HNSCC. In both acute and prolonged hypoxia (1% oxygen), the B7H3 TriKE mediated enhanced tumor killing, mitigating impairment of NK cell cytotoxicity in hypoxia. In vivo, the B7H3 TriKE-treated mice demonstrated substantial antitumor activity and prolonged survival.Conclusions The B7H3 TriKE is a novel immunotherapeutic approach that can overcome hypoxic suppression of NK cells in the HNSCC TME. These highly translational studies present an innovative therapy for patients with HNSCC and will be developed further for clinical application.
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spelling doaj-art-95da99fe87f74bcf98d15b362697cb9b2025-08-20T03:13:43ZengBMJ Publishing GroupJournal for ImmunoTherapy of Cancer2051-14262025-07-0113710.1136/jitc-2024-011370Novel trispecific killer engager targeting B7-H3 enhances natural killer cell antitumor activity against head and neck cancerJeffrey S Miller0Martin Felices1Peter Hinderlie2James Lim3Naomi Fujioka4Shee Kwan Phung5Yvette Soignier6Joshua T Walker7Melissa A Geller8Madison Shackelford9Carly Selleck10Philippa R Kennedy11Mihir Shetty12Melissa J Khaw13Paolo P Provenzano14Nicholas A Zorko15Laura E Bendzick16Riley C Lyons17Elise L Femino18Terran Stenger19Tumpa Dasgupta20Laura E Kotz21Eng Hock Lee22Quynhanh Lu23Zachary B Davis24John E Wagner25Margaret L MacMillan262 Department of Medicine, University of Minnesota, Minneapolis, Minnesota, USA2 Department of Medicine, University of Minnesota, Minneapolis, Minnesota, USA1 University of Minnesota Masonic Cancer Center, Minneapolis, Minnesota, USA5 Xcell Biosciences, San Francisco, California, USA1 University of Minnesota Masonic Cancer Center, Minneapolis, Minnesota, USA1 University of Minnesota Masonic Cancer Center, Minneapolis, Minnesota, USA1 University of Minnesota Masonic Cancer Center, Minneapolis, Minnesota, USA1 University of Minnesota Masonic Cancer Center, Minneapolis, Minnesota, USA1 University of Minnesota Masonic Cancer Center, Minneapolis, Minnesota, USA1 University of Minnesota Masonic Cancer Center, Minneapolis, Minnesota, USA1 University of Minnesota Masonic Cancer Center, Minneapolis, Minnesota, USA1 University of Minnesota Masonic Cancer Center, Minneapolis, Minnesota, USA1 University of Minnesota Masonic Cancer Center, Minneapolis, Minnesota, USA1 University of Minnesota Masonic Cancer Center, Minneapolis, Minnesota, USA6 Department of Biomedical Engineering, University of Minnesota Medical School Twin Cities, Minneapolis, Minnesota, USA1 University of Minnesota Masonic Cancer Center, Minneapolis, Minnesota, USA1 University of Minnesota Masonic Cancer Center, Minneapolis, Minnesota, USA1 University of Minnesota Masonic Cancer Center, Minneapolis, Minnesota, USA1 University of Minnesota Masonic Cancer Center, Minneapolis, Minnesota, USA1 University of Minnesota Masonic Cancer Center, Minneapolis, Minnesota, USA1 University of Minnesota Masonic Cancer Center, Minneapolis, Minnesota, USA1 University of Minnesota Masonic Cancer Center, Minneapolis, Minnesota, USA4 Department of Electrical and Computer Engineering, University of Minnesota, Minneapolis, Minnesota, USA5 Xcell Biosciences, San Francisco, California, USA1 University of Minnesota Masonic Cancer Center, Minneapolis, Minnesota, USA1 University of Minnesota Masonic Cancer Center, Minneapolis, Minnesota, USA1 University of Minnesota Masonic Cancer Center, Minneapolis, Minnesota, USABackground Patients with head and neck squamous cell carcinoma (HNSCC), particularly the human papillomavirus negative (HPV−) subset, have a dismal prognosis. Furthermore, patients with Fanconi anemia (FA) have a genetic predisposition with a 500-fold to 700-fold higher incidence of HNSCC. Thus, novel and more efficacious therapies are needed. As current immunotherapies often fail due to suppressive elements in the tumor microenvironment (TME), we developed a trispecific killer engager (TriKE) to direct multiple signals to natural killer (NK) cells to overcome the hypoxic TME. This TriKE is comprised of a camelid nanobody that binds to CD16 on NK cells, an interleukin (IL)-15 moiety, and another novel camelid nanobody that binds to the B7-H3 antigen, which is highly and specifically expressed on the tumor cell surface.Methods The B7H3 TriKE was generated using a mammalian expression system. Its functionality was evaluated using flow cytometry-based NK cell degranulation, cytokine production, proliferation and live cell imaging cytotoxicity assays. Models of acute and prolonged hypoxia (1% oxygen) were carried out to assess tumor killing. Tumor progression, NK cell persistence, and survival differences between IL-15-treated and TriKE-treated mice were studied using NOD-scidIL2Rgnull (NSG) mice engrafted with human HNSCC.Results High B7-H3 expression was found in HPV− HNSCC cell lines, even when the FA gene was knocked out, and The Cancer Genome Atlas patient data showed that high B7-H3 expression predicted poor survival in patients with HPV− HNSCC. Similar to the NK cell activity seen with healthy donors, the B7H3 TriKE enhanced activation, expansion and cytotoxicity of NK cells from patients with HPV− HNSCC, a target population for this therapeutic. Additionally, the B7H3 TriKE improved NK cell cytotoxicity in a three-dimensional spheroid model of HNSCC. In both acute and prolonged hypoxia (1% oxygen), the B7H3 TriKE mediated enhanced tumor killing, mitigating impairment of NK cell cytotoxicity in hypoxia. In vivo, the B7H3 TriKE-treated mice demonstrated substantial antitumor activity and prolonged survival.Conclusions The B7H3 TriKE is a novel immunotherapeutic approach that can overcome hypoxic suppression of NK cells in the HNSCC TME. These highly translational studies present an innovative therapy for patients with HNSCC and will be developed further for clinical application.https://jitc.bmj.com/content/13/7/e011370.full
spellingShingle Jeffrey S Miller
Martin Felices
Peter Hinderlie
James Lim
Naomi Fujioka
Shee Kwan Phung
Yvette Soignier
Joshua T Walker
Melissa A Geller
Madison Shackelford
Carly Selleck
Philippa R Kennedy
Mihir Shetty
Melissa J Khaw
Paolo P Provenzano
Nicholas A Zorko
Laura E Bendzick
Riley C Lyons
Elise L Femino
Terran Stenger
Tumpa Dasgupta
Laura E Kotz
Eng Hock Lee
Quynhanh Lu
Zachary B Davis
John E Wagner
Margaret L MacMillan
Novel trispecific killer engager targeting B7-H3 enhances natural killer cell antitumor activity against head and neck cancer
Journal for ImmunoTherapy of Cancer
title Novel trispecific killer engager targeting B7-H3 enhances natural killer cell antitumor activity against head and neck cancer
title_full Novel trispecific killer engager targeting B7-H3 enhances natural killer cell antitumor activity against head and neck cancer
title_fullStr Novel trispecific killer engager targeting B7-H3 enhances natural killer cell antitumor activity against head and neck cancer
title_full_unstemmed Novel trispecific killer engager targeting B7-H3 enhances natural killer cell antitumor activity against head and neck cancer
title_short Novel trispecific killer engager targeting B7-H3 enhances natural killer cell antitumor activity against head and neck cancer
title_sort novel trispecific killer engager targeting b7 h3 enhances natural killer cell antitumor activity against head and neck cancer
url https://jitc.bmj.com/content/13/7/e011370.full
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