Research ArticleImmunologyOncology Open Access | 10.1172/jci.insight.178767

Chang Su,1,2 Collin L. Kent,2 Matthew Pierpoint,2 Warren Floyd,3 Lixia Luo,2 Nerissa T. Williams,2 Yan Ma,2 Brian Peng,2 Alexander L. Lazarides,2 Ajay Subramanian,4 Jonathon E. Himes,1,2 Vincent M. Perez,5 Rosa D. Hernansaiz-Ballesteros,5 Kimberly E. Roche,5,6 Jennifer L. Modliszewski,6,7 Sara R. Selitsky,5,6 Mari L. Shinohara,8,9,10 Amy J. Wisdom,11 Everett J. Moding,4,12 Yvonne M. Mowery,2,13 and David G. Kirsch1,2,14,15,16

1Department of Pharmacology and Cancer Biology and

2Department of Radiation Oncology, Duke University Medical Center, Durham, North Carolina, USA.

3Department of Radiation Oncology, MD Anderson Cancer Center, Houston, Texas, USA.

4Department of Radiation Oncology, Stanford University, Stanford, California, USA.

5Tempus AI Inc., Durham, North Carolina, USA.

6QuantBio LLC, Durham, North Carolina, USA.

7Lineberger Comprehensive Cancer Center, Chapel Hill, North Carolina, USA.

8Department of Integrative Immunology,

9Department of Molecular Genetics and Microbiology, and

10Department of Neurobiology, Duke University School of Medicine, Durham, North Carolina, USA.

11Harvard Radiation Oncology Program, Boston, Massachusetts, USA.

12Stanford Cancer Institute, Stanford University, Stanford, California, USA.

13Department of Radiation Oncology, University of Pittsburgh, Pittsburgh, Pennsylvania, USA.

14Department of Radiation Oncology and

15Department of Medical Biophysics, University of Toronto, Toronto, Ontario, Canada.

16Radiation Medicine Program, Princess Margaret Cancer Centre, University Health Network, Toronto, Ontario, Canada.

Address correspondence to: David G. Kirsch, 610 University Ave., Toronto, Ontario M5G 2M9, Canada. Phone: 416.946.2094; Email: David.Kirsch@uhn.ca.

Authorship note: CS and CLK are co–first authors.

Find articles by Su, C. in: JCI | PubMed | Google Scholar |

1Department of Pharmacology and Cancer Biology and

2Department of Radiation Oncology, Duke University Medical Center, Durham, North Carolina, USA.

3Department of Radiation Oncology, MD Anderson Cancer Center, Houston, Texas, USA.

4Department of Radiation Oncology, Stanford University, Stanford, California, USA.

5Tempus AI Inc., Durham, North Carolina, USA.

6QuantBio LLC, Durham, North Carolina, USA.

7Lineberger Comprehensive Cancer Center, Chapel Hill, North Carolina, USA.

8Department of Integrative Immunology,

9Department of Molecular Genetics and Microbiology, and

10Department of Neurobiology, Duke University School of Medicine, Durham, North Carolina, USA.

11Harvard Radiation Oncology Program, Boston, Massachusetts, USA.

12Stanford Cancer Institute, Stanford University, Stanford, California, USA.

13Department of Radiation Oncology, University of Pittsburgh, Pittsburgh, Pennsylvania, USA.

14Department of Radiation Oncology and

15Department of Medical Biophysics, University of Toronto, Toronto, Ontario, Canada.

16Radiation Medicine Program, Princess Margaret Cancer Centre, University Health Network, Toronto, Ontario, Canada.

Address correspondence to: David G. Kirsch, 610 University Ave., Toronto, Ontario M5G 2M9, Canada. Phone: 416.946.2094; Email: David.Kirsch@uhn.ca.

Authorship note: CS and CLK are co–first authors.

Find articles by Kent, C. in: JCI | PubMed | Google Scholar

1Department of Pharmacology and Cancer Biology and

2Department of Radiation Oncology, Duke University Medical Center, Durham, North Carolina, USA.

3Department of Radiation Oncology, MD Anderson Cancer Center, Houston, Texas, USA.

4Department of Radiation Oncology, Stanford University, Stanford, California, USA.

5Tempus AI Inc., Durham, North Carolina, USA.

6QuantBio LLC, Durham, North Carolina, USA.

7Lineberger Comprehensive Cancer Center, Chapel Hill, North Carolina, USA.

8Department of Integrative Immunology,

9Department of Molecular Genetics and Microbiology, and

10Department of Neurobiology, Duke University School of Medicine, Durham, North Carolina, USA.

11Harvard Radiation Oncology Program, Boston, Massachusetts, USA.

12Stanford Cancer Institute, Stanford University, Stanford, California, USA.

13Department of Radiation Oncology, University of Pittsburgh, Pittsburgh, Pennsylvania, USA.

14Department of Radiation Oncology and

15Department of Medical Biophysics, University of Toronto, Toronto, Ontario, Canada.

16Radiation Medicine Program, Princess Margaret Cancer Centre, University Health Network, Toronto, Ontario, Canada.

Address correspondence to: David G. Kirsch, 610 University Ave., Toronto, Ontario M5G 2M9, Canada. Phone: 416.946.2094; Email: David.Kirsch@uhn.ca.

Authorship note: CS and CLK are co–first authors.

Find articles by Pierpoint, M. in: JCI | PubMed | Google Scholar

1Department of Pharmacology and Cancer Biology and

2Department of Radiation Oncology, Duke University Medical Center, Durham, North Carolina, USA.

3Department of Radiation Oncology, MD Anderson Cancer Center, Houston, Texas, USA.

4Department of Radiation Oncology, Stanford University, Stanford, California, USA.

5Tempus AI Inc., Durham, North Carolina, USA.

6QuantBio LLC, Durham, North Carolina, USA.

7Lineberger Comprehensive Cancer Center, Chapel Hill, North Carolina, USA.

8Department of Integrative Immunology,

9Department of Molecular Genetics and Microbiology, and

10Department of Neurobiology, Duke University School of Medicine, Durham, North Carolina, USA.

11Harvard Radiation Oncology Program, Boston, Massachusetts, USA.

12Stanford Cancer Institute, Stanford University, Stanford, California, USA.

13Department of Radiation Oncology, University of Pittsburgh, Pittsburgh, Pennsylvania, USA.

14Department of Radiation Oncology and

15Department of Medical Biophysics, University of Toronto, Toronto, Ontario, Canada.

16Radiation Medicine Program, Princess Margaret Cancer Centre, University Health Network, Toronto, Ontario, Canada.

Address correspondence to: David G. Kirsch, 610 University Ave., Toronto, Ontario M5G 2M9, Canada. Phone: 416.946.2094; Email: David.Kirsch@uhn.ca.

Authorship note: CS and CLK are co–first authors.

Find articles by Floyd, W. in: JCI | PubMed | Google Scholar

1Department of Pharmacology and Cancer Biology and

2Department of Radiation Oncology, Duke University Medical Center, Durham, North Carolina, USA.

3Department of Radiation Oncology, MD Anderson Cancer Center, Houston, Texas, USA.

4Department of Radiation Oncology, Stanford University, Stanford, California, USA.

5Tempus AI Inc., Durham, North Carolina, USA.

6QuantBio LLC, Durham, North Carolina, USA.

7Lineberger Comprehensive Cancer Center, Chapel Hill, North Carolina, USA.

8Department of Integrative Immunology,

9Department of Molecular Genetics and Microbiology, and

10Department of Neurobiology, Duke University School of Medicine, Durham, North Carolina, USA.

11Harvard Radiation Oncology Program, Boston, Massachusetts, USA.

12Stanford Cancer Institute, Stanford University, Stanford, California, USA.

13Department of Radiation Oncology, University of Pittsburgh, Pittsburgh, Pennsylvania, USA.

14Department of Radiation Oncology and

15Department of Medical Biophysics, University of Toronto, Toronto, Ontario, Canada.

16Radiation Medicine Program, Princess Margaret Cancer Centre, University Health Network, Toronto, Ontario, Canada.

Address correspondence to: David G. Kirsch, 610 University Ave., Toronto, Ontario M5G 2M9, Canada. Phone: 416.946.2094; Email: David.Kirsch@uhn.ca.

Authorship note: CS and CLK are co–first authors.

Find articles by Luo, L. in: JCI | PubMed | Google Scholar

1Department of Pharmacology and Cancer Biology and

2Department of Radiation Oncology, Duke University Medical Center, Durham, North Carolina, USA.

3Department of Radiation Oncology, MD Anderson Cancer Center, Houston, Texas, USA.

4Department of Radiation Oncology, Stanford University, Stanford, California, USA.

5Tempus AI Inc., Durham, North Carolina, USA.

6QuantBio LLC, Durham, North Carolina, USA.

7Lineberger Comprehensive Cancer Center, Chapel Hill, North Carolina, USA.

8Department of Integrative Immunology,

9Department of Molecular Genetics and Microbiology, and

10Department of Neurobiology, Duke University School of Medicine, Durham, North Carolina, USA.

11Harvard Radiation Oncology Program, Boston, Massachusetts, USA.

12Stanford Cancer Institute, Stanford University, Stanford, California, USA.

13Department of Radiation Oncology, University of Pittsburgh, Pittsburgh, Pennsylvania, USA.

14Department of Radiation Oncology and

15Department of Medical Biophysics, University of Toronto, Toronto, Ontario, Canada.

16Radiation Medicine Program, Princess Margaret Cancer Centre, University Health Network, Toronto, Ontario, Canada.

Address correspondence to: David G. Kirsch, 610 University Ave., Toronto, Ontario M5G 2M9, Canada. Phone: 416.946.2094; Email: David.Kirsch@uhn.ca.

Authorship note: CS and CLK are co–first authors.

Find articles by Williams, N. in: JCI | PubMed | Google Scholar

1Department of Pharmacology and Cancer Biology and

2Department of Radiation Oncology, Duke University Medical Center, Durham, North Carolina, USA.

3Department of Radiation Oncology, MD Anderson Cancer Center, Houston, Texas, USA.

4Department of Radiation Oncology, Stanford University, Stanford, California, USA.

5Tempus AI Inc., Durham, North Carolina, USA.

6QuantBio LLC, Durham, North Carolina, USA.

7Lineberger Comprehensive Cancer Center, Chapel Hill, North Carolina, USA.

8Department of Integrative Immunology,

9Department of Molecular Genetics and Microbiology, and

10Department of Neurobiology, Duke University School of Medicine, Durham, North Carolina, USA.

11Harvard Radiation Oncology Program, Boston, Massachusetts, USA.

12Stanford Cancer Institute, Stanford University, Stanford, California, USA.

13Department of Radiation Oncology, University of Pittsburgh, Pittsburgh, Pennsylvania, USA.

14Department of Radiation Oncology and

15Department of Medical Biophysics, University of Toronto, Toronto, Ontario, Canada.

16Radiation Medicine Program, Princess Margaret Cancer Centre, University Health Network, Toronto, Ontario, Canada.

Address correspondence to: David G. Kirsch, 610 University Ave., Toronto, Ontario M5G 2M9, Canada. Phone: 416.946.2094; Email: David.Kirsch@uhn.ca.

Authorship note: CS and CLK are co–first authors.

Find articles by Ma, Y. in: JCI | PubMed | Google Scholar

1Department of Pharmacology and Cancer Biology and

2Department of Radiation Oncology, Duke University Medical Center, Durham, North Carolina, USA.

3Department of Radiation Oncology, MD Anderson Cancer Center, Houston, Texas, USA.

4Department of Radiation Oncology, Stanford University, Stanford, California, USA.

5Tempus AI Inc., Durham, North Carolina, USA.

6QuantBio LLC, Durham, North Carolina, USA.

7Lineberger Comprehensive Cancer Center, Chapel Hill, North Carolina, USA.

8Department of Integrative Immunology,

9Department of Molecular Genetics and Microbiology, and

10Department of Neurobiology, Duke University School of Medicine, Durham, North Carolina, USA.

11Harvard Radiation Oncology Program, Boston, Massachusetts, USA.

12Stanford Cancer Institute, Stanford University, Stanford, California, USA.

13Department of Radiation Oncology, University of Pittsburgh, Pittsburgh, Pennsylvania, USA.

14Department of Radiation Oncology and

15Department of Medical Biophysics, University of Toronto, Toronto, Ontario, Canada.

16Radiation Medicine Program, Princess Margaret Cancer Centre, University Health Network, Toronto, Ontario, Canada.

Address correspondence to: David G. Kirsch, 610 University Ave., Toronto, Ontario M5G 2M9, Canada. Phone: 416.946.2094; Email: David.Kirsch@uhn.ca.

Authorship note: CS and CLK are co–first authors.

Find articles by Peng, B. in: JCI | PubMed | Google Scholar

1Department of Pharmacology and Cancer Biology and

2Department of Radiation Oncology, Duke University Medical Center, Durham, North Carolina, USA.

3Department of Radiation Oncology, MD Anderson Cancer Center, Houston, Texas, USA.

4Department of Radiation Oncology, Stanford University, Stanford, California, USA.

5Tempus AI Inc., Durham, North Carolina, USA.

6QuantBio LLC, Durham, North Carolina, USA.

7Lineberger Comprehensive Cancer Center, Chapel Hill, North Carolina, USA.

8Department of Integrative Immunology,

9Department of Molecular Genetics and Microbiology, and

10Department of Neurobiology, Duke University School of Medicine, Durham, North Carolina, USA.

11Harvard Radiation Oncology Program, Boston, Massachusetts, USA.

12Stanford Cancer Institute, Stanford University, Stanford, California, USA.

13Department of Radiation Oncology, University of Pittsburgh, Pittsburgh, Pennsylvania, USA.

14Department of Radiation Oncology and

15Department of Medical Biophysics, University of Toronto, Toronto, Ontario, Canada.

16Radiation Medicine Program, Princess Margaret Cancer Centre, University Health Network, Toronto, Ontario, Canada.

Address correspondence to: David G. Kirsch, 610 University Ave., Toronto, Ontario M5G 2M9, Canada. Phone: 416.946.2094; Email: David.Kirsch@uhn.ca.

Authorship note: CS and CLK are co–first authors.

Find articles by Lazarides, A. in: JCI | PubMed | Google Scholar |

1Department of Pharmacology and Cancer Biology and

2Department of Radiation Oncology, Duke University Medical Center, Durham, North Carolina, USA.

3Department of Radiation Oncology, MD Anderson Cancer Center, Houston, Texas, USA.

4Department of Radiation Oncology, Stanford University, Stanford, California, USA.

5Tempus AI Inc., Durham, North Carolina, USA.

6QuantBio LLC, Durham, North Carolina, USA.

7Lineberger Comprehensive Cancer Center, Chapel Hill, North Carolina, USA.

8Department of Integrative Immunology,

9Department of Molecular Genetics and Microbiology, and

10Department of Neurobiology, Duke University School of Medicine, Durham, North Carolina, USA.

11Harvard Radiation Oncology Program, Boston, Massachusetts, USA.

12Stanford Cancer Institute, Stanford University, Stanford, California, USA.

13Department of Radiation Oncology, University of Pittsburgh, Pittsburgh, Pennsylvania, USA.

14Department of Radiation Oncology and

15Department of Medical Biophysics, University of Toronto, Toronto, Ontario, Canada.

16Radiation Medicine Program, Princess Margaret Cancer Centre, University Health Network, Toronto, Ontario, Canada.

Address correspondence to: David G. Kirsch, 610 University Ave., Toronto, Ontario M5G 2M9, Canada. Phone: 416.946.2094; Email: David.Kirsch@uhn.ca.

Authorship note: CS and CLK are co–first authors.

Find articles by Subramanian, A. in: JCI | PubMed | Google Scholar |

1Department of Pharmacology and Cancer Biology and

2Department of Radiation Oncology, Duke University Medical Center, Durham, North Carolina, USA.

3Department of Radiation Oncology, MD Anderson Cancer Center, Houston, Texas, USA.

4Department of Radiation Oncology, Stanford University, Stanford, California, USA.

5Tempus AI Inc., Durham, North Carolina, USA.

6QuantBio LLC, Durham, North Carolina, USA.

7Lineberger Comprehensive Cancer Center, Chapel Hill, North Carolina, USA.

8Department of Integrative Immunology,

9Department of Molecular Genetics and Microbiology, and

10Department of Neurobiology, Duke University School of Medicine, Durham, North Carolina, USA.

11Harvard Radiation Oncology Program, Boston, Massachusetts, USA.

12Stanford Cancer Institute, Stanford University, Stanford, California, USA.

13Department of Radiation Oncology, University of Pittsburgh, Pittsburgh, Pennsylvania, USA.

14Department of Radiation Oncology and

15Department of Medical Biophysics, University of Toronto, Toronto, Ontario, Canada.

16Radiation Medicine Program, Princess Margaret Cancer Centre, University Health Network, Toronto, Ontario, Canada.

Address correspondence to: David G. Kirsch, 610 University Ave., Toronto, Ontario M5G 2M9, Canada. Phone: 416.946.2094; Email: David.Kirsch@uhn.ca.

Authorship note: CS and CLK are co–first authors.

Find articles by Himes, J. in: JCI | PubMed | Google Scholar

1Department of Pharmacology and Cancer Biology and

2Department of Radiation Oncology, Duke University Medical Center, Durham, North Carolina, USA.

3Department of Radiation Oncology, MD Anderson Cancer Center, Houston, Texas, USA.

4Department of Radiation Oncology, Stanford University, Stanford, California, USA.

5Tempus AI Inc., Durham, North Carolina, USA.

6QuantBio LLC, Durham, North Carolina, USA.

7Lineberger Comprehensive Cancer Center, Chapel Hill, North Carolina, USA.

8Department of Integrative Immunology,

9Department of Molecular Genetics and Microbiology, and

10Department of Neurobiology, Duke University School of Medicine, Durham, North Carolina, USA.

11Harvard Radiation Oncology Program, Boston, Massachusetts, USA.

12Stanford Cancer Institute, Stanford University, Stanford, California, USA.

13Department of Radiation Oncology, University of Pittsburgh, Pittsburgh, Pennsylvania, USA.

14Department of Radiation Oncology and

15Department of Medical Biophysics, University of Toronto, Toronto, Ontario, Canada.

16Radiation Medicine Program, Princess Margaret Cancer Centre, University Health Network, Toronto, Ontario, Canada.

Address correspondence to: David G. Kirsch, 610 University Ave., Toronto, Ontario M5G 2M9, Canada. Phone: 416.946.2094; Email: David.Kirsch@uhn.ca.

Authorship note: CS and CLK are co–first authors.

Find articles by Perez, V. in: JCI | PubMed | Google Scholar

1Department of Pharmacology and Cancer Biology and

2Department of Radiation Oncology, Duke University Medical Center, Durham, North Carolina, USA.

3Department of Radiation Oncology, MD Anderson Cancer Center, Houston, Texas, USA.

4Department of Radiation Oncology, Stanford University, Stanford, California, USA.

5Tempus AI Inc., Durham, North Carolina, USA.

6QuantBio LLC, Durham, North Carolina, USA.

7Lineberger Comprehensive Cancer Center, Chapel Hill, North Carolina, USA.

8Department of Integrative Immunology,

9Department of Molecular Genetics and Microbiology, and

10Department of Neurobiology, Duke University School of Medicine, Durham, North Carolina, USA.

11Harvard Radiation Oncology Program, Boston, Massachusetts, USA.

12Stanford Cancer Institute, Stanford University, Stanford, California, USA.

13Department of Radiation Oncology, University of Pittsburgh, Pittsburgh, Pennsylvania, USA.

14Department of Radiation Oncology and

15Department of Medical Biophysics, University of Toronto, Toronto, Ontario, Canada.

16Radiation Medicine Program, Princess Margaret Cancer Centre, University Health Network, Toronto, Ontario, Canada.

Address correspondence to: David G. Kirsch, 610 University Ave., Toronto, Ontario M5G 2M9, Canada. Phone: 416.946.2094; Email: David.Kirsch@uhn.ca.

Authorship note: CS and CLK are co–first authors.

Find articles by Hernansaiz-Ballesteros, R. in: JCI | PubMed | Google Scholar

1Department of Pharmacology and Cancer Biology and

2Department of Radiation Oncology, Duke University Medical Center, Durham, North Carolina, USA.

3Department of Radiation Oncology, MD Anderson Cancer Center, Houston, Texas, USA.

4Department of Radiation Oncology, Stanford University, Stanford, California, USA.

5Tempus AI Inc., Durham, North Carolina, USA.

6QuantBio LLC, Durham, North Carolina, USA.

7Lineberger Comprehensive Cancer Center, Chapel Hill, North Carolina, USA.

8Department of Integrative Immunology,

9Department of Molecular Genetics and Microbiology, and

10Department of Neurobiology, Duke University School of Medicine, Durham, North Carolina, USA.

11Harvard Radiation Oncology Program, Boston, Massachusetts, USA.

12Stanford Cancer Institute, Stanford University, Stanford, California, USA.

13Department of Radiation Oncology, University of Pittsburgh, Pittsburgh, Pennsylvania, USA.

14Department of Radiation Oncology and

15Department of Medical Biophysics, University of Toronto, Toronto, Ontario, Canada.

16Radiation Medicine Program, Princess Margaret Cancer Centre, University Health Network, Toronto, Ontario, Canada.

Address correspondence to: David G. Kirsch, 610 University Ave., Toronto, Ontario M5G 2M9, Canada. Phone: 416.946.2094; Email: David.Kirsch@uhn.ca.

Authorship note: CS and CLK are co–first authors.

Find articles by Roche, K. in: JCI | PubMed | Google Scholar

1Department of Pharmacology and Cancer Biology and

2Department of Radiation Oncology, Duke University Medical Center, Durham, North Carolina, USA.

3Department of Radiation Oncology, MD Anderson Cancer Center, Houston, Texas, USA.

4Department of Radiation Oncology, Stanford University, Stanford, California, USA.

5Tempus AI Inc., Durham, North Carolina, USA.

6QuantBio LLC, Durham, North Carolina, USA.

7Lineberger Comprehensive Cancer Center, Chapel Hill, North Carolina, USA.

8Department of Integrative Immunology,

9Department of Molecular Genetics and Microbiology, and

10Department of Neurobiology, Duke University School of Medicine, Durham, North Carolina, USA.

11Harvard Radiation Oncology Program, Boston, Massachusetts, USA.

12Stanford Cancer Institute, Stanford University, Stanford, California, USA.

13Department of Radiation Oncology, University of Pittsburgh, Pittsburgh, Pennsylvania, USA.

14Department of Radiation Oncology and

15Department of Medical Biophysics, University of Toronto, Toronto, Ontario, Canada.

16Radiation Medicine Program, Princess Margaret Cancer Centre, University Health Network, Toronto, Ontario, Canada.

Address correspondence to: David G. Kirsch, 610 University Ave., Toronto, Ontario M5G 2M9, Canada. Phone: 416.946.2094; Email: David.Kirsch@uhn.ca.

Authorship note: CS and CLK are co–first authors.

Find articles by Modliszewski, J. in: JCI | PubMed | Google Scholar

1Department of Pharmacology and Cancer Biology and

2Department of Radiation Oncology, Duke University Medical Center, Durham, North Carolina, USA.

3Department of Radiation Oncology, MD Anderson Cancer Center, Houston, Texas, USA.

4Department of Radiation Oncology, Stanford University, Stanford, California, USA.

5Tempus AI Inc., Durham, North Carolina, USA.

6QuantBio LLC, Durham, North Carolina, USA.

7Lineberger Comprehensive Cancer Center, Chapel Hill, North Carolina, USA.

8Department of Integrative Immunology,

9Department of Molecular Genetics and Microbiology, and

10Department of Neurobiology, Duke University School of Medicine, Durham, North Carolina, USA.

11Harvard Radiation Oncology Program, Boston, Massachusetts, USA.

12Stanford Cancer Institute, Stanford University, Stanford, California, USA.

13Department of Radiation Oncology, University of Pittsburgh, Pittsburgh, Pennsylvania, USA.

14Department of Radiation Oncology and

15Department of Medical Biophysics, University of Toronto, Toronto, Ontario, Canada.

16Radiation Medicine Program, Princess Margaret Cancer Centre, University Health Network, Toronto, Ontario, Canada.

Address correspondence to: David G. Kirsch, 610 University Ave., Toronto, Ontario M5G 2M9, Canada. Phone: 416.946.2094; Email: David.Kirsch@uhn.ca.

Authorship note: CS and CLK are co–first authors.

Find articles by Selitsky, S. in: JCI | PubMed | Google Scholar

1Department of Pharmacology and Cancer Biology and

2Department of Radiation Oncology, Duke University Medical Center, Durham, North Carolina, USA.

3Department of Radiation Oncology, MD Anderson Cancer Center, Houston, Texas, USA.

4Department of Radiation Oncology, Stanford University, Stanford, California, USA.

5Tempus AI Inc., Durham, North Carolina, USA.

6QuantBio LLC, Durham, North Carolina, USA.

7Lineberger Comprehensive Cancer Center, Chapel Hill, North Carolina, USA.

8Department of Integrative Immunology,

9Department of Molecular Genetics and Microbiology, and

10Department of Neurobiology, Duke University School of Medicine, Durham, North Carolina, USA.

11Harvard Radiation Oncology Program, Boston, Massachusetts, USA.

12Stanford Cancer Institute, Stanford University, Stanford, California, USA.

13Department of Radiation Oncology, University of Pittsburgh, Pittsburgh, Pennsylvania, USA.

14Department of Radiation Oncology and

15Department of Medical Biophysics, University of Toronto, Toronto, Ontario, Canada.

16Radiation Medicine Program, Princess Margaret Cancer Centre, University Health Network, Toronto, Ontario, Canada.

Address correspondence to: David G. Kirsch, 610 University Ave., Toronto, Ontario M5G 2M9, Canada. Phone: 416.946.2094; Email: David.Kirsch@uhn.ca.

Authorship note: CS and CLK are co–first authors.

Find articles by Shinohara, M. in: JCI | PubMed | Google Scholar |

1Department of Pharmacology and Cancer Biology and

2Department of Radiation Oncology, Duke University Medical Center, Durham, North Carolina, USA.

3Department of Radiation Oncology, MD Anderson Cancer Center, Houston, Texas, USA.

4Department of Radiation Oncology, Stanford University, Stanford, California, USA.

5Tempus AI Inc., Durham, North Carolina, USA.

6QuantBio LLC, Durham, North Carolina, USA.

7Lineberger Comprehensive Cancer Center, Chapel Hill, North Carolina, USA.

8Department of Integrative Immunology,

9Department of Molecular Genetics and Microbiology, and

10Department of Neurobiology, Duke University School of Medicine, Durham, North Carolina, USA.

11Harvard Radiation Oncology Program, Boston, Massachusetts, USA.

12Stanford Cancer Institute, Stanford University, Stanford, California, USA.

13Department of Radiation Oncology, University of Pittsburgh, Pittsburgh, Pennsylvania, USA.

14Department of Radiation Oncology and

15Department of Medical Biophysics, University of Toronto, Toronto, Ontario, Canada.

16Radiation Medicine Program, Princess Margaret Cancer Centre, University Health Network, Toronto, Ontario, Canada.

Address correspondence to: David G. Kirsch, 610 University Ave., Toronto, Ontario M5G 2M9, Canada. Phone: 416.946.2094; Email: David.Kirsch@uhn.ca.

Authorship note: CS and CLK are co–first authors.

Find articles by Wisdom, A. in: JCI | PubMed | Google Scholar |

1Department of Pharmacology and Cancer Biology and

2Department of Radiation Oncology, Duke University Medical Center, Durham, North Carolina, USA.

3Department of Radiation Oncology, MD Anderson Cancer Center, Houston, Texas, USA.

4Department of Radiation Oncology, Stanford University, Stanford, California, USA.

5Tempus AI Inc., Durham, North Carolina, USA.

6QuantBio LLC, Durham, North Carolina, USA.

7Lineberger Comprehensive Cancer Center, Chapel Hill, North Carolina, USA.

8Department of Integrative Immunology,

9Department of Molecular Genetics and Microbiology, and

10Department of Neurobiology, Duke University School of Medicine, Durham, North Carolina, USA.

11Harvard Radiation Oncology Program, Boston, Massachusetts, USA.

12Stanford Cancer Institute, Stanford University, Stanford, California, USA.

13Department of Radiation Oncology, University of Pittsburgh, Pittsburgh, Pennsylvania, USA.

14Department of Radiation Oncology and

15Department of Medical Biophysics, University of Toronto, Toronto, Ontario, Canada.

16Radiation Medicine Program, Princess Margaret Cancer Centre, University Health Network, Toronto, Ontario, Canada.

Address correspondence to: David G. Kirsch, 610 University Ave., Toronto, Ontario M5G 2M9, Canada. Phone: 416.946.2094; Email: David.Kirsch@uhn.ca.

Authorship note: CS and CLK are co–first authors.

Find articles by Moding, E. in: JCI | PubMed | Google Scholar

1Department of Pharmacology and Cancer Biology and

2Department of Radiation Oncology, Duke University Medical Center, Durham, North Carolina, USA.

3Department of Radiation Oncology, MD Anderson Cancer Center, Houston, Texas, USA.

4Department of Radiation Oncology, Stanford University, Stanford, California, USA.

5Tempus AI Inc., Durham, North Carolina, USA.

6QuantBio LLC, Durham, North Carolina, USA.

7Lineberger Comprehensive Cancer Center, Chapel Hill, North Carolina, USA.

8Department of Integrative Immunology,

9Department of Molecular Genetics and Microbiology, and

10Department of Neurobiology, Duke University School of Medicine, Durham, North Carolina, USA.

11Harvard Radiation Oncology Program, Boston, Massachusetts, USA.

12Stanford Cancer Institute, Stanford University, Stanford, California, USA.

13Department of Radiation Oncology, University of Pittsburgh, Pittsburgh, Pennsylvania, USA.

14Department of Radiation Oncology and

15Department of Medical Biophysics, University of Toronto, Toronto, Ontario, Canada.

16Radiation Medicine Program, Princess Margaret Cancer Centre, University Health Network, Toronto, Ontario, Canada.

Address correspondence to: David G. Kirsch, 610 University Ave., Toronto, Ontario M5G 2M9, Canada. Phone: 416.946.2094; Email: David.Kirsch@uhn.ca.

Authorship note: CS and CLK are co–first authors.

Find articles by Mowery, Y. in: JCI | PubMed | Google Scholar |

1Department of Pharmacology and Cancer Biology and

2Department of Radiation Oncology, Duke University Medical Center, Durham, North Carolina, USA.

3Department of Radiation Oncology, MD Anderson Cancer Center, Houston, Texas, USA.

4Department of Radiation Oncology, Stanford University, Stanford, California, USA.

5Tempus AI Inc., Durham, North Carolina, USA.

6QuantBio LLC, Durham, North Carolina, USA.

7Lineberger Comprehensive Cancer Center, Chapel Hill, North Carolina, USA.

8Department of Integrative Immunology,

9Department of Molecular Genetics and Microbiology, and

10Department of Neurobiology, Duke University School of Medicine, Durham, North Carolina, USA.

11Harvard Radiation Oncology Program, Boston, Massachusetts, USA.

12Stanford Cancer Institute, Stanford University, Stanford, California, USA.

13Department of Radiation Oncology, University of Pittsburgh, Pittsburgh, Pennsylvania, USA.

14Department of Radiation Oncology and

15Department of Medical Biophysics, University of Toronto, Toronto, Ontario, Canada.

16Radiation Medicine Program, Princess Margaret Cancer Centre, University Health Network, Toronto, Ontario, Canada.

Address correspondence to: David G. Kirsch, 610 University Ave., Toronto, Ontario M5G 2M9, Canada. Phone: 416.946.2094; Email: David.Kirsch@uhn.ca.

Authorship note: CS and CLK are co–first authors.

Find articles by Kirsch, D. in: JCI | PubMed | Google Scholar |

Authorship note: CS and CLK are co–first authors.

Published July 22, 2024 - More info

Published in Volume 9, Issue 14 on July 22, 2024
JCI Insight. 2024;9(14):e178767. https://doi.org/10.1172/jci.insight.178767.
© 2024 Su et al. This work is licensed under the Creative Commons Attribution 4.0 International License. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/. Published July 22, 2024 - Version history
Received: January 3, 2024; Accepted: June 11, 2024 View PDF Abstract

Radiation therapy (RT) is frequently used to treat cancers, including soft-tissue sarcomas. Prior studies established that the toll-like receptor 9 (TLR9) agonist cytosine-phosphate-guanine oligodeoxynucleotide (CpG) enhances the response to RT in transplanted tumors, but the mechanisms of this enhancement remain unclear. Here, we used CRISPR/Cas9 and the chemical carcinogen 3-methylcholanthrene (MCA) to generate autochthonous soft-tissue sarcomas with high tumor mutation burden. Treatment with a single fraction of 20 Gy RT and 2 doses of CpG significantly enhanced tumor response, which was abrogated by genetic or immunodepletion of CD8+ T cells. To characterize the immune response to CpG+RT, we performed bulk RNA-Seq, single-cell RNA-Seq, and mass cytometry. Sarcomas treated with 20 Gy and CpG demonstrated increased CD8 T cells expressing markers associated with activation and proliferation, such as Granzyme B, Ki-67, and IFN-γ. CpG+RT also upregulated antigen presentation pathways on myeloid cells. Furthermore, in sarcomas treated with CpG+RT, TCR clonality analysis suggests an increase in clonal T cell dominance. Collectively, these findings demonstrate that CpG+RT significantly delays tumor growth in a CD8 T cell–dependent manner. These results provide a strong rationale for clinical trials evaluating CpG or other TLR9 agonists with RT in patients with soft-tissue sarcoma.

Graphical Abstract Introduction

Soft-tissue sarcomas (STS) are a heterogeneous group of malignancies (1). Following lo