Targeting axonal guidance dependencies in glioblastoma with ROBO1 CAR T cells

Chirayu R Chokshi^{1

2}, Muhammad Vaseem Shaikh^#^{2

3}, Benjamin Brakel^#^{1

2}, Martin A Rossotti⁴, David Tieu^{5

6}, William Maich^{1

2}, Alisha Anand^{1

2}, Shawn C Chafe^{2

3}, Kui Zhai^{2

3}, Yujin Suk^{1

2}, Agata M Kieliszek^{1

2}, Petar Miletic^{2

3}, Nicholas Mikolajewicz⁷, David Chen⁷, Jamie D McNicol^{8

9}, Katherine Chan⁷, Amy H Y Tong⁵, Laura Kuhlmann¹⁰, Lina Liu¹, Zahra Alizada^{2

3}, Daniel Mobilio^{1

2}, Nazanin Tatari^{1

2}, Neil Savage^{1

2}, Nikoo Aghaei^{1

2}, Shan Grewal^{1

2}, Anish Puri^{1

2}, Minomi Subapanditha⁹, Dillon McKenna^{2

3}, Vladimir Ignatchenko¹⁰, Joseph M Salamoun¹¹, Jacek M Kwiecien¹², Peter Wipf¹¹, Elizabeth R Sharlow¹³, John P Provias¹², Jian-Qiang Lu¹², John S Lazo¹³, Thomas Kislinger^{10

14}, Yu Lu², Kevin R Brown⁷, Chitra Venugopal^{2

3}, Kevin A Henry^{4

15}, Jason Moffat^{16

17

18}, Sheila K Singh^{19

20

21}

Affiliations

¹ Department of Biochemistry and Biomedical Sciences, McMaster University, Hamilton, ON, Canada.
² Centre for Discovery in Cancer Research (CDCR), McMaster University, Hamilton, ON, Canada.
³ Department of Surgery, Faculty of Health Sciences, McMaster University, Hamilton, ON, Canada.
⁴ Human Health Therapeutics Research Centre, Life Sciences Division, National Research Council Canada, Ottawa, ON, Canada.
⁵ Donnelly Centre, University of Toronto, Toronto, ON, Canada.
⁶ Department of Molecular Genetics, University of Toronto, Toronto, ON, Canada.
⁷ Program for Genetics and Genome Biology, The Hospital for Sick Children, Peter Gilgan Centre for Research & Learning, Toronto, ON, Canada.
⁸ McMaster Immunology Research Centre, McMaster University, Hamilton, ON, Canada.
⁹ Department of Medicine, McMaster University, Hamilton, ON, Canada.
¹⁰ Princess Margaret Cancer Centre, University Health Network, Toronto, ON, Canada.
¹¹ Department of Chemistry, University of Pittsburgh, Pittsburgh, PA, USA.
¹² Department of Pathology and Molecular Medicine, Faculty of Health Sciences, McMaster University, Hamilton, ON, Canada.
¹³ Department of Pharmacology, Fiske Drug Discovery Laboratory, University of Virginia, Charlottesville, VA, USA.
¹⁴ Department of Medical Biophysics, University of Toronto, Toronto, ON, Canada.
¹⁵ Department of Biochemistry, Microbiology and Immunology, Faculty of Medicine, University of Ottawa, Ottawa, ON, Canada.
¹⁶ Department of Molecular Genetics, University of Toronto, Toronto, ON, Canada. jason.moffat@sickkids.ca.
¹⁷ Program for Genetics and Genome Biology, The Hospital for Sick Children, Peter Gilgan Centre for Research & Learning, Toronto, ON, Canada. jason.moffat@sickkids.ca.
¹⁸ Institute for Biomaterials and Biomedical Engineering, University of Toronto, Toronto, ON, Canada. jason.moffat@sickkids.ca.
¹⁹ Department of Biochemistry and Biomedical Sciences, McMaster University, Hamilton, ON, Canada. ssingh@mcmaster.ca.
²⁰ Centre for Discovery in Cancer Research (CDCR), McMaster University, Hamilton, ON, Canada. ssingh@mcmaster.ca.
²¹ Department of Surgery, Faculty of Health Sciences, McMaster University, Hamilton, ON, Canada. ssingh@mcmaster.ca.

^# Contributed equally.

PMID: 39095594
DOI: 10.1038/s41591-024-03138-9

Targeting axonal guidance dependencies in glioblastoma with ROBO1 CAR T cells

Chirayu R Chokshi et al. Nat Med. 2024 Oct.

. 2024 Oct;30(10):2936-2946.

doi: 10.1038/s41591-024-03138-9. Epub 2024 Aug 2.

Authors

Affiliations

¹ Department of Biochemistry and Biomedical Sciences, McMaster University, Hamilton, ON, Canada.
² Centre for Discovery in Cancer Research (CDCR), McMaster University, Hamilton, ON, Canada.
³ Department of Surgery, Faculty of Health Sciences, McMaster University, Hamilton, ON, Canada.
⁴ Human Health Therapeutics Research Centre, Life Sciences Division, National Research Council Canada, Ottawa, ON, Canada.
⁵ Donnelly Centre, University of Toronto, Toronto, ON, Canada.
⁶ Department of Molecular Genetics, University of Toronto, Toronto, ON, Canada.
⁷ Program for Genetics and Genome Biology, The Hospital for Sick Children, Peter Gilgan Centre for Research & Learning, Toronto, ON, Canada.
⁸ McMaster Immunology Research Centre, McMaster University, Hamilton, ON, Canada.
⁹ Department of Medicine, McMaster University, Hamilton, ON, Canada.
¹⁰ Princess Margaret Cancer Centre, University Health Network, Toronto, ON, Canada.
¹¹ Department of Chemistry, University of Pittsburgh, Pittsburgh, PA, USA.
¹² Department of Pathology and Molecular Medicine, Faculty of Health Sciences, McMaster University, Hamilton, ON, Canada.
¹³ Department of Pharmacology, Fiske Drug Discovery Laboratory, University of Virginia, Charlottesville, VA, USA.
¹⁴ Department of Medical Biophysics, University of Toronto, Toronto, ON, Canada.
¹⁵ Department of Biochemistry, Microbiology and Immunology, Faculty of Medicine, University of Ottawa, Ottawa, ON, Canada.
¹⁶ Department of Molecular Genetics, University of Toronto, Toronto, ON, Canada. jason.moffat@sickkids.ca.
¹⁷ Program for Genetics and Genome Biology, The Hospital for Sick Children, Peter Gilgan Centre for Research & Learning, Toronto, ON, Canada. jason.moffat@sickkids.ca.
¹⁸ Institute for Biomaterials and Biomedical Engineering, University of Toronto, Toronto, ON, Canada. jason.moffat@sickkids.ca.
¹⁹ Department of Biochemistry and Biomedical Sciences, McMaster University, Hamilton, ON, Canada. ssingh@mcmaster.ca.
²⁰ Centre for Discovery in Cancer Research (CDCR), McMaster University, Hamilton, ON, Canada. ssingh@mcmaster.ca.
²¹ Department of Surgery, Faculty of Health Sciences, McMaster University, Hamilton, ON, Canada. ssingh@mcmaster.ca.

^# Contributed equally.

PMID: 39095594
DOI: 10.1038/s41591-024-03138-9

Abstract

Resistance to genotoxic therapies and tumor recurrence are hallmarks of glioblastoma (GBM), an aggressive brain tumor. In this study, we investigated functional drivers of post-treatment recurrent GBM through integrative genomic analyses, genome-wide genetic perturbation screens in patient-derived GBM models and independent lines of validation. Specific genetic dependencies were found consistent across recurrent tumor models, accompanied by increased mutational burden and differential transcript and protein expression compared to its primary GBM predecessor. Our observations suggest a multi-layered genetic response to drive tumor recurrence and implicate PTP4A2 (protein tyrosine phosphatase 4A2) as a modulator of self-renewal, proliferation and tumorigenicity in recurrent GBM. Genetic perturbation or small-molecule inhibition of PTP4A2 acts through a dephosphorylation axis with roundabout guidance receptor 1 (ROBO1) and its downstream molecular players, exploiting a functional dependency on ROBO signaling. Because a pan-PTP4A inhibitor was limited by poor penetrance across the blood-brain barrier in vivo, we engineered a second-generation chimeric antigen receptor (CAR) T cell therapy against ROBO1, a cell surface receptor enriched across recurrent GBM specimens. A single dose of ROBO1-targeted CAR T cells doubled median survival in cell-line-derived xenograft (CDX) models of recurrent GBM. Moreover, in CDX models of adult lung-to-brain metastases and pediatric relapsed medulloblastoma, ROBO1 CAR T cells eradicated tumors in 50-100% of mice. Our study identifies a promising multi-targetable PTP4A-ROBO1 signaling axis that drives tumorigenicity in recurrent GBM, with potential in other malignant brain tumors.

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References

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Targeting axonal guidance dependencies in glioblastoma with ROBO1 CAR T cells

Affiliations

Targeting axonal guidance dependencies in glioblastoma with ROBO1 CAR T cells

Authors

Affiliations

Abstract

References

MeSH terms

Substances

Grants and funding

LinkOut - more resources

Full Text Sources

Medical

Molecular Biology Databases