2. Academic Validation
  3. Development of a high-affinity anti-ROR1 variable region for broad anti-cancer immunotherapy

Development of a high-affinity anti-ROR1 variable region for broad anti-cancer immunotherapy

  • Mol Ther. 2025 Dec 2:S1525-0016(25)00959-1. doi: 10.1016/j.ymthe.2025.11.021.
Joshua K M Wong 1,  Pui Yeng Lam 1,  Elaina Coleborn 1,  Jiya Jose 1,  Louisa Alim 1,  Cui Tu 1,  Magdalena Antczak 2,  Bettina Dietmair 3,  Arghavan Golbaz Hagh 4,  Lucia Noronha 5,  Seth W Cheetham 3,  John Hooper 6,  Paul A Beavis 7,  Delphine Merino 8,  Jean Berthelet 8,  Lauren G Aoude 1,  Amy E McCart-Reed 9,  Sunil Lakhani 9,  Peter T Simpson 10,  Gustavo R Rossi 1,  Andrew J Brooks 11,  Martina L Jones 4,  Fiona Simpson 1,  Fernando Souza-Fonseca-Guimaraes 12
Affiliations

Affiliations

  • 1 Frazer Institute, The University of Queensland, Woolloongabba, QLD 4102, Australia.
  • 2 Queensland Cyber Infrastructure Foundation Ltd (QCIF) Bioinformatics, Brisbane, QLD 4072, Australia.
  • 3 Australian Institute for Bioengineering and Nanotechnology, University of Queensland, St Lucia, QLD 4067, Australia; BASE Facility, University of Queensland, St Lucia, QLD 4067, Australia.
  • 4 Australian Institute for Bioengineering and Nanotechnology, University of Queensland, St Lucia, QLD 4067, Australia.
  • 5 Laboratório de Patologia Experimental, Curitiba, Queensland 80215-901, Brazil.
  • 6 Mater Research Institute, The University of Queensland, Brisbane, QLD 4102, Australia.
  • 7 Cancer Immunology Program, Peter MacCallum Cancer Centre, Melbourne, VIC 3000, Australia; Sir Peter MacCallum Department of Oncology, The University of Melbourne, Parkville, VIC 3010, Australia.
  • 8 Olivia Newton-John Cancer Research Institute, Heidelberg, VIC 3084, Australia.
  • 9 UQ Centre for Clinical Research, Faculty of Health, Medicine and Behavioural Sciences, The University of Queensland, Herston, QLD 4029, Australia.
  • 10 UQ Centre for Clinical Research, Faculty of Health, Medicine and Behavioural Sciences, The University of Queensland, Herston, QLD 4029, Australia; School of Biomedical Sciences, Faculty of Health, Medicine and Behavioural Sciences, The University of Queensland, Saint Lucia, QLD 4067, Australia.
  • 11 Frazer Institute, The University of Queensland, Woolloongabba, QLD 4102, Australia; School of Science & Technology, University of New England, Armidale NSW 2351, Australia.
  • 12 Frazer Institute, The University of Queensland, Woolloongabba, QLD 4102, Australia. Electronic address: [email protected].
PMID: 41338184 DOI: 10.1016/j.ymthe.2025.11.021
Abstract

Receptor tyrosine kinase-like Orphan Receptor 1 (ROR1) is an emerging target in Cancer Immunotherapy, recognized for its consistent and elevated expression across several epithelial tumors, including triple-negative breast Cancer (TNBC). TNBC is an aggressive and difficult-to-treat Cancer, with limited effective therapeutic options currently available. Therapeutic approaches centered on targeting ROR1 have therefore become increasingly popular, with ROR1 chimeric antigen receptor (CAR) T cells currently in clinical trials to treat TNBC patients. While ROR1-targeting therapies have shown promising preclinical results, single arm treatment has often shown low efficacy as well as off-target toxicity. Natural killer (NK) cell-based immunotherapies, such as antibody-dependent cell cytotoxicity-inducing monoclonal antibodies and CAR NK cells, have also been shown to induce Cancer cell cytotoxicity; however, with less toxicity compared with CAR T cells. Here, we developed and characterized a phage-derived single-chain fragment variable (scFv) against a highly specific ROR1 region and generated scFv-derived chimeric monoclonal antibodies and anti-ROR1-CAR NK cells, which show anti-cancer efficacy against TNBC cells. Additionally, we found TGF-β inhibition using either small-molecule inhibitors or CRISPR-Cas9-edited NK cells could further enhance ROR1-targeting therapy persistence and efficacy in controlling TNBC tumor growth.

Keywords

ADCC; CAR NK cells; CRISPR/Cas9; Frizzled-Kringle domain; TGFBR2 knockout; antibody drug conjugate; natural killer cells; phage display; scFv; triple negative breast cancer.

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