Research ArticleOncologyTherapeutics Open Access | 10.1172/JCI174794

Jun Watanabe,1,2,3,4 Matthew R. Clutter,5 Michael J. Gullette,6 Takahiro Sasaki,7,8 Eita Uchida,1,2,3 Savneet Kaur,3 Yan Mo,9,10,11 Kouki Abe,1,2 Yukitomo Ishi,1,2 Nozomu Takata,12,13 Manabu Natsumeda,4 Samantha Gadd,2 Zhiguo Zhang,9,10,11 Oren J. Becher,14 and Rintaro Hashizume1,2,3,15,16

1Department of Pediatrics, Northwestern University Feinberg School of Medicine, Chicago, Illinois, USA.

2Division of Hematology, Oncology, Neuro-Oncology and Stem Cell Transplantation, Ann & Robert H. Lurie Children’s Hospital of Chicago, Chicago, Illinois, USA.

3Department of Pediatrics, University of Alabama at Birmingham, Birmingham, Alabama, USA.

4Department of Neurological Surgery, Brain Research Institute, Niigata University, Niigata, Japan.

5Department of Molecular Biosciences,

6High Throughput Analysis Laboratory, and

7Department of Neurological Surgery, Northwestern University Feinberg School of Medicine, Chicago, Illinois, USA.

8Department of Neurological Surgery, Wakayama Medical University, Wakayama, Japan.

9Institute for Cancer Genetics,

10Department of Pediatrics, and

11Department of Genetics and Development, Columbia University Irving Medical Center, New York, New York, USA.

12Center for Vascular and Developmental Biology, Feinberg Cardiovascular and Renal Research Institute, and

13Simpson Querrey Institute for BioNanotechnology, Northwestern University Feinberg School of Medicine, Chicago, Illinois, USA.

14Department of Pediatrics, Icahn School of Medicine at Mount Sinai, New York, New York, USA.

15Division of Pediatric Hematology and Oncology, Children’s of Alabama, Birmingham, Alabama, USA.

16O’Neal Comprehensive Cancer Center, University of Alabama at Birmingham, Birmingham, Alabama, USA.

Address correspondence to: Oren J. Becher, Department of Pediatrics, Icahn School of Medicine at Mount Sinai, 1468 Madison Avenue, 4th Floor, New York, New York 10029, USA. Phone: 212.241.7022; Email: oren.becher@mssm.edu. Or to: Rintaro Hashizume, Department of Pediatrics, University of Alabama at Birmingham, 1670 University Boulevard, G019A, Birmingham, Alabama 35294, USA. Phone: 205.975.0285; Email: rhashi23@uab.edu.

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

1Department of Pediatrics, Northwestern University Feinberg School of Medicine, Chicago, Illinois, USA.

2Division of Hematology, Oncology, Neuro-Oncology and Stem Cell Transplantation, Ann & Robert H. Lurie Children’s Hospital of Chicago, Chicago, Illinois, USA.

3Department of Pediatrics, University of Alabama at Birmingham, Birmingham, Alabama, USA.

4Department of Neurological Surgery, Brain Research Institute, Niigata University, Niigata, Japan.

5Department of Molecular Biosciences,

6High Throughput Analysis Laboratory, and

7Department of Neurological Surgery, Northwestern University Feinberg School of Medicine, Chicago, Illinois, USA.

8Department of Neurological Surgery, Wakayama Medical University, Wakayama, Japan.

9Institute for Cancer Genetics,

10Department of Pediatrics, and

11Department of Genetics and Development, Columbia University Irving Medical Center, New York, New York, USA.

12Center for Vascular and Developmental Biology, Feinberg Cardiovascular and Renal Research Institute, and

13Simpson Querrey Institute for BioNanotechnology, Northwestern University Feinberg School of Medicine, Chicago, Illinois, USA.

14Department of Pediatrics, Icahn School of Medicine at Mount Sinai, New York, New York, USA.

15Division of Pediatric Hematology and Oncology, Children’s of Alabama, Birmingham, Alabama, USA.

16O’Neal Comprehensive Cancer Center, University of Alabama at Birmingham, Birmingham, Alabama, USA.

Address correspondence to: Oren J. Becher, Department of Pediatrics, Icahn School of Medicine at Mount Sinai, 1468 Madison Avenue, 4th Floor, New York, New York 10029, USA. Phone: 212.241.7022; Email: oren.becher@mssm.edu. Or to: Rintaro Hashizume, Department of Pediatrics, University of Alabama at Birmingham, 1670 University Boulevard, G019A, Birmingham, Alabama 35294, USA. Phone: 205.975.0285; Email: rhashi23@uab.edu.

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

1Department of Pediatrics, Northwestern University Feinberg School of Medicine, Chicago, Illinois, USA.

2Division of Hematology, Oncology, Neuro-Oncology and Stem Cell Transplantation, Ann & Robert H. Lurie Children’s Hospital of Chicago, Chicago, Illinois, USA.

3Department of Pediatrics, University of Alabama at Birmingham, Birmingham, Alabama, USA.

4Department of Neurological Surgery, Brain Research Institute, Niigata University, Niigata, Japan.

5Department of Molecular Biosciences,

6High Throughput Analysis Laboratory, and

7Department of Neurological Surgery, Northwestern University Feinberg School of Medicine, Chicago, Illinois, USA.

8Department of Neurological Surgery, Wakayama Medical University, Wakayama, Japan.

9Institute for Cancer Genetics,

10Department of Pediatrics, and

11Department of Genetics and Development, Columbia University Irving Medical Center, New York, New York, USA.

12Center for Vascular and Developmental Biology, Feinberg Cardiovascular and Renal Research Institute, and

13Simpson Querrey Institute for BioNanotechnology, Northwestern University Feinberg School of Medicine, Chicago, Illinois, USA.

14Department of Pediatrics, Icahn School of Medicine at Mount Sinai, New York, New York, USA.

15Division of Pediatric Hematology and Oncology, Children’s of Alabama, Birmingham, Alabama, USA.

16O’Neal Comprehensive Cancer Center, University of Alabama at Birmingham, Birmingham, Alabama, USA.

Address correspondence to: Oren J. Becher, Department of Pediatrics, Icahn School of Medicine at Mount Sinai, 1468 Madison Avenue, 4th Floor, New York, New York 10029, USA. Phone: 212.241.7022; Email: oren.becher@mssm.edu. Or to: Rintaro Hashizume, Department of Pediatrics, University of Alabama at Birmingham, 1670 University Boulevard, G019A, Birmingham, Alabama 35294, USA. Phone: 205.975.0285; Email: rhashi23@uab.edu.

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

1Department of Pediatrics, Northwestern University Feinberg School of Medicine, Chicago, Illinois, USA.

2Division of Hematology, Oncology, Neuro-Oncology and Stem Cell Transplantation, Ann & Robert H. Lurie Children’s Hospital of Chicago, Chicago, Illinois, USA.

3Department of Pediatrics, University of Alabama at Birmingham, Birmingham, Alabama, USA.

4Department of Neurological Surgery, Brain Research Institute, Niigata University, Niigata, Japan.

5Department of Molecular Biosciences,

6High Throughput Analysis Laboratory, and

7Department of Neurological Surgery, Northwestern University Feinberg School of Medicine, Chicago, Illinois, USA.

8Department of Neurological Surgery, Wakayama Medical University, Wakayama, Japan.

9Institute for Cancer Genetics,

10Department of Pediatrics, and

11Department of Genetics and Development, Columbia University Irving Medical Center, New York, New York, USA.

12Center for Vascular and Developmental Biology, Feinberg Cardiovascular and Renal Research Institute, and

13Simpson Querrey Institute for BioNanotechnology, Northwestern University Feinberg School of Medicine, Chicago, Illinois, USA.

14Department of Pediatrics, Icahn School of Medicine at Mount Sinai, New York, New York, USA.

15Division of Pediatric Hematology and Oncology, Children’s of Alabama, Birmingham, Alabama, USA.

16O’Neal Comprehensive Cancer Center, University of Alabama at Birmingham, Birmingham, Alabama, USA.

Address correspondence to: Oren J. Becher, Department of Pediatrics, Icahn School of Medicine at Mount Sinai, 1468 Madison Avenue, 4th Floor, New York, New York 10029, USA. Phone: 212.241.7022; Email: oren.becher@mssm.edu. Or to: Rintaro Hashizume, Department of Pediatrics, University of Alabama at Birmingham, 1670 University Boulevard, G019A, Birmingham, Alabama 35294, USA. Phone: 205.975.0285; Email: rhashi23@uab.edu.

Find articles by Sasaki, T. in: JCI | PubMed | Google Scholar

1Department of Pediatrics, Northwestern University Feinberg School of Medicine, Chicago, Illinois, USA.

2Division of Hematology, Oncology, Neuro-Oncology and Stem Cell Transplantation, Ann & Robert H. Lurie Children’s Hospital of Chicago, Chicago, Illinois, USA.

3Department of Pediatrics, University of Alabama at Birmingham, Birmingham, Alabama, USA.

4Department of Neurological Surgery, Brain Research Institute, Niigata University, Niigata, Japan.

5Department of Molecular Biosciences,

6High Throughput Analysis Laboratory, and

7Department of Neurological Surgery, Northwestern University Feinberg School of Medicine, Chicago, Illinois, USA.

8Department of Neurological Surgery, Wakayama Medical University, Wakayama, Japan.

9Institute for Cancer Genetics,

10Department of Pediatrics, and

11Department of Genetics and Development, Columbia University Irving Medical Center, New York, New York, USA.

12Center for Vascular and Developmental Biology, Feinberg Cardiovascular and Renal Research Institute, and

13Simpson Querrey Institute for BioNanotechnology, Northwestern University Feinberg School of Medicine, Chicago, Illinois, USA.

14Department of Pediatrics, Icahn School of Medicine at Mount Sinai, New York, New York, USA.

15Division of Pediatric Hematology and Oncology, Children’s of Alabama, Birmingham, Alabama, USA.

16O’Neal Comprehensive Cancer Center, University of Alabama at Birmingham, Birmingham, Alabama, USA.

Address correspondence to: Oren J. Becher, Department of Pediatrics, Icahn School of Medicine at Mount Sinai, 1468 Madison Avenue, 4th Floor, New York, New York 10029, USA. Phone: 212.241.7022; Email: oren.becher@mssm.edu. Or to: Rintaro Hashizume, Department of Pediatrics, University of Alabama at Birmingham, 1670 University Boulevard, G019A, Birmingham, Alabama 35294, USA. Phone: 205.975.0285; Email: rhashi23@uab.edu.

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

1Department of Pediatrics, Northwestern University Feinberg School of Medicine, Chicago, Illinois, USA.

2Division of Hematology, Oncology, Neuro-Oncology and Stem Cell Transplantation, Ann & Robert H. Lurie Children’s Hospital of Chicago, Chicago, Illinois, USA.

3Department of Pediatrics, University of Alabama at Birmingham, Birmingham, Alabama, USA.

4Department of Neurological Surgery, Brain Research Institute, Niigata University, Niigata, Japan.

5Department of Molecular Biosciences,

6High Throughput Analysis Laboratory, and

7Department of Neurological Surgery, Northwestern University Feinberg School of Medicine, Chicago, Illinois, USA.

8Department of Neurological Surgery, Wakayama Medical University, Wakayama, Japan.

9Institute for Cancer Genetics,

10Department of Pediatrics, and

11Department of Genetics and Development, Columbia University Irving Medical Center, New York, New York, USA.

12Center for Vascular and Developmental Biology, Feinberg Cardiovascular and Renal Research Institute, and

13Simpson Querrey Institute for BioNanotechnology, Northwestern University Feinberg School of Medicine, Chicago, Illinois, USA.

14Department of Pediatrics, Icahn School of Medicine at Mount Sinai, New York, New York, USA.

15Division of Pediatric Hematology and Oncology, Children’s of Alabama, Birmingham, Alabama, USA.

16O’Neal Comprehensive Cancer Center, University of Alabama at Birmingham, Birmingham, Alabama, USA.

Address correspondence to: Oren J. Becher, Department of Pediatrics, Icahn School of Medicine at Mount Sinai, 1468 Madison Avenue, 4th Floor, New York, New York 10029, USA. Phone: 212.241.7022; Email: oren.becher@mssm.edu. Or to: Rintaro Hashizume, Department of Pediatrics, University of Alabama at Birmingham, 1670 University Boulevard, G019A, Birmingham, Alabama 35294, USA. Phone: 205.975.0285; Email: rhashi23@uab.edu.

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

1Department of Pediatrics, Northwestern University Feinberg School of Medicine, Chicago, Illinois, USA.

2Division of Hematology, Oncology, Neuro-Oncology and Stem Cell Transplantation, Ann & Robert H. Lurie Children’s Hospital of Chicago, Chicago, Illinois, USA.

3Department of Pediatrics, University of Alabama at Birmingham, Birmingham, Alabama, USA.

4Department of Neurological Surgery, Brain Research Institute, Niigata University, Niigata, Japan.

5Department of Molecular Biosciences,

6High Throughput Analysis Laboratory, and

7Department of Neurological Surgery, Northwestern University Feinberg School of Medicine, Chicago, Illinois, USA.

8Department of Neurological Surgery, Wakayama Medical University, Wakayama, Japan.

9Institute for Cancer Genetics,

10Department of Pediatrics, and

11Department of Genetics and Development, Columbia University Irving Medical Center, New York, New York, USA.

12Center for Vascular and Developmental Biology, Feinberg Cardiovascular and Renal Research Institute, and

13Simpson Querrey Institute for BioNanotechnology, Northwestern University Feinberg School of Medicine, Chicago, Illinois, USA.

14Department of Pediatrics, Icahn School of Medicine at Mount Sinai, New York, New York, USA.

15Division of Pediatric Hematology and Oncology, Children’s of Alabama, Birmingham, Alabama, USA.

16O’Neal Comprehensive Cancer Center, University of Alabama at Birmingham, Birmingham, Alabama, USA.

Address correspondence to: Oren J. Becher, Department of Pediatrics, Icahn School of Medicine at Mount Sinai, 1468 Madison Avenue, 4th Floor, New York, New York 10029, USA. Phone: 212.241.7022; Email: oren.becher@mssm.edu. Or to: Rintaro Hashizume, Department of Pediatrics, University of Alabama at Birmingham, 1670 University Boulevard, G019A, Birmingham, Alabama 35294, USA. Phone: 205.975.0285; Email: rhashi23@uab.edu.

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

1Department of Pediatrics, Northwestern University Feinberg School of Medicine, Chicago, Illinois, USA.

2Division of Hematology, Oncology, Neuro-Oncology and Stem Cell Transplantation, Ann & Robert H. Lurie Children’s Hospital of Chicago, Chicago, Illinois, USA.

3Department of Pediatrics, University of Alabama at Birmingham, Birmingham, Alabama, USA.

4Department of Neurological Surgery, Brain Research Institute, Niigata University, Niigata, Japan.

5Department of Molecular Biosciences,

6High Throughput Analysis Laboratory, and

7Department of Neurological Surgery, Northwestern University Feinberg School of Medicine, Chicago, Illinois, USA.

8Department of Neurological Surgery, Wakayama Medical University, Wakayama, Japan.

9Institute for Cancer Genetics,

10Department of Pediatrics, and

11Department of Genetics and Development, Columbia University Irving Medical Center, New York, New York, USA.

12Center for Vascular and Developmental Biology, Feinberg Cardiovascular and Renal Research Institute, and

13Simpson Querrey Institute for BioNanotechnology, Northwestern University Feinberg School of Medicine, Chicago, Illinois, USA.

14Department of Pediatrics, Icahn School of Medicine at Mount Sinai, New York, New York, USA.

15Division of Pediatric Hematology and Oncology, Children’s of Alabama, Birmingham, Alabama, USA.

16O’Neal Comprehensive Cancer Center, University of Alabama at Birmingham, Birmingham, Alabama, USA.

Address correspondence to: Oren J. Becher, Department of Pediatrics, Icahn School of Medicine at Mount Sinai, 1468 Madison Avenue, 4th Floor, New York, New York 10029, USA. Phone: 212.241.7022; Email: oren.becher@mssm.edu. Or to: Rintaro Hashizume, Department of Pediatrics, University of Alabama at Birmingham, 1670 University Boulevard, G019A, Birmingham, Alabama 35294, USA. Phone: 205.975.0285; Email: rhashi23@uab.edu.

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

1Department of Pediatrics, Northwestern University Feinberg School of Medicine, Chicago, Illinois, USA.

2Division of Hematology, Oncology, Neuro-Oncology and Stem Cell Transplantation, Ann & Robert H. Lurie Children’s Hospital of Chicago, Chicago, Illinois, USA.

3Department of Pediatrics, University of Alabama at Birmingham, Birmingham, Alabama, USA.

4Department of Neurological Surgery, Brain Research Institute, Niigata University, Niigata, Japan.

5Department of Molecular Biosciences,

6High Throughput Analysis Laboratory, and

7Department of Neurological Surgery, Northwestern University Feinberg School of Medicine, Chicago, Illinois, USA.

8Department of Neurological Surgery, Wakayama Medical University, Wakayama, Japan.

9Institute for Cancer Genetics,

10Department of Pediatrics, and

11Department of Genetics and Development, Columbia University Irving Medical Center, New York, New York, USA.

12Center for Vascular and Developmental Biology, Feinberg Cardiovascular and Renal Research Institute, and

13Simpson Querrey Institute for BioNanotechnology, Northwestern University Feinberg School of Medicine, Chicago, Illinois, USA.

14Department of Pediatrics, Icahn School of Medicine at Mount Sinai, New York, New York, USA.

15Division of Pediatric Hematology and Oncology, Children’s of Alabama, Birmingham, Alabama, USA.

16O’Neal Comprehensive Cancer Center, University of Alabama at Birmingham, Birmingham, Alabama, USA.

Address correspondence to: Oren J. Becher, Department of Pediatrics, Icahn School of Medicine at Mount Sinai, 1468 Madison Avenue, 4th Floor, New York, New York 10029, USA. Phone: 212.241.7022; Email: oren.becher@mssm.edu. Or to: Rintaro Hashizume, Department of Pediatrics, University of Alabama at Birmingham, 1670 University Boulevard, G019A, Birmingham, Alabama 35294, USA. Phone: 205.975.0285; Email: rhashi23@uab.edu.

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

1Department of Pediatrics, Northwestern University Feinberg School of Medicine, Chicago, Illinois, USA.

2Division of Hematology, Oncology, Neuro-Oncology and Stem Cell Transplantation, Ann & Robert H. Lurie Children’s Hospital of Chicago, Chicago, Illinois, USA.

3Department of Pediatrics, University of Alabama at Birmingham, Birmingham, Alabama, USA.

4Department of Neurological Surgery, Brain Research Institute, Niigata University, Niigata, Japan.

5Department of Molecular Biosciences,

6High Throughput Analysis Laboratory, and

7Department of Neurological Surgery, Northwestern University Feinberg School of Medicine, Chicago, Illinois, USA.

8Department of Neurological Surgery, Wakayama Medical University, Wakayama, Japan.

9Institute for Cancer Genetics,

10Department of Pediatrics, and

11Department of Genetics and Development, Columbia University Irving Medical Center, New York, New York, USA.

12Center for Vascular and Developmental Biology, Feinberg Cardiovascular and Renal Research Institute, and

13Simpson Querrey Institute for BioNanotechnology, Northwestern University Feinberg School of Medicine, Chicago, Illinois, USA.

14Department of Pediatrics, Icahn School of Medicine at Mount Sinai, New York, New York, USA.

15Division of Pediatric Hematology and Oncology, Children’s of Alabama, Birmingham, Alabama, USA.

16O’Neal Comprehensive Cancer Center, University of Alabama at Birmingham, Birmingham, Alabama, USA.

Address correspondence to: Oren J. Becher, Department of Pediatrics, Icahn School of Medicine at Mount Sinai, 1468 Madison Avenue, 4th Floor, New York, New York 10029, USA. Phone: 212.241.7022; Email: oren.becher@mssm.edu. Or to: Rintaro Hashizume, Department of Pediatrics, University of Alabama at Birmingham, 1670 University Boulevard, G019A, Birmingham, Alabama 35294, USA. Phone: 205.975.0285; Email: rhashi23@uab.edu.

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

1Department of Pediatrics, Northwestern University Feinberg School of Medicine, Chicago, Illinois, USA.

2Division of Hematology, Oncology, Neuro-Oncology and Stem Cell Transplantation, Ann & Robert H. Lurie Children’s Hospital of Chicago, Chicago, Illinois, USA.

3Department of Pediatrics, University of Alabama at Birmingham, Birmingham, Alabama, USA.

4Department of Neurological Surgery, Brain Research Institute, Niigata University, Niigata, Japan.

5Department of Molecular Biosciences,

6High Throughput Analysis Laboratory, and

7Department of Neurological Surgery, Northwestern University Feinberg School of Medicine, Chicago, Illinois, USA.

8Department of Neurological Surgery, Wakayama Medical University, Wakayama, Japan.

9Institute for Cancer Genetics,

10Department of Pediatrics, and

11Department of Genetics and Development, Columbia University Irving Medical Center, New York, New York, USA.

12Center for Vascular and Developmental Biology, Feinberg Cardiovascular and Renal Research Institute, and

13Simpson Querrey Institute for BioNanotechnology, Northwestern University Feinberg School of Medicine, Chicago, Illinois, USA.

14Department of Pediatrics, Icahn School of Medicine at Mount Sinai, New York, New York, USA.

15Division of Pediatric Hematology and Oncology, Children’s of Alabama, Birmingham, Alabama, USA.

16O’Neal Comprehensive Cancer Center, University of Alabama at Birmingham, Birmingham, Alabama, USA.

Address correspondence to: Oren J. Becher, Department of Pediatrics, Icahn School of Medicine at Mount Sinai, 1468 Madison Avenue, 4th Floor, New York, New York 10029, USA. Phone: 212.241.7022; Email: oren.becher@mssm.edu. Or to: Rintaro Hashizume, Department of Pediatrics, University of Alabama at Birmingham, 1670 University Boulevard, G019A, Birmingham, Alabama 35294, USA. Phone: 205.975.0285; Email: rhashi23@uab.edu.

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

1Department of Pediatrics, Northwestern University Feinberg School of Medicine, Chicago, Illinois, USA.

2Division of Hematology, Oncology, Neuro-Oncology and Stem Cell Transplantation, Ann & Robert H. Lurie Children’s Hospital of Chicago, Chicago, Illinois, USA.

3Department of Pediatrics, University of Alabama at Birmingham, Birmingham, Alabama, USA.

4Department of Neurological Surgery, Brain Research Institute, Niigata University, Niigata, Japan.

5Department of Molecular Biosciences,

6High Throughput Analysis Laboratory, and

7Department of Neurological Surgery, Northwestern University Feinberg School of Medicine, Chicago, Illinois, USA.

8Department of Neurological Surgery, Wakayama Medical University, Wakayama, Japan.

9Institute for Cancer Genetics,

10Department of Pediatrics, and

11Department of Genetics and Development, Columbia University Irving Medical Center, New York, New York, USA.

12Center for Vascular and Developmental Biology, Feinberg Cardiovascular and Renal Research Institute, and

13Simpson Querrey Institute for BioNanotechnology, Northwestern University Feinberg School of Medicine, Chicago, Illinois, USA.

14Department of Pediatrics, Icahn School of Medicine at Mount Sinai, New York, New York, USA.

15Division of Pediatric Hematology and Oncology, Children’s of Alabama, Birmingham, Alabama, USA.

16O’Neal Comprehensive Cancer Center, University of Alabama at Birmingham, Birmingham, Alabama, USA.

Address correspondence to: Oren J. Becher, Department of Pediatrics, Icahn School of Medicine at Mount Sinai, 1468 Madison Avenue, 4th Floor, New York, New York 10029, USA. Phone: 212.241.7022; Email: oren.becher@mssm.edu. Or to: Rintaro Hashizume, Department of Pediatrics, University of Alabama at Birmingham, 1670 University Boulevard, G019A, Birmingham, Alabama 35294, USA. Phone: 205.975.0285; Email: rhashi23@uab.edu.

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

1Department of Pediatrics, Northwestern University Feinberg School of Medicine, Chicago, Illinois, USA.

2Division of Hematology, Oncology, Neuro-Oncology and Stem Cell Transplantation, Ann & Robert H. Lurie Children’s Hospital of Chicago, Chicago, Illinois, USA.

3Department of Pediatrics, University of Alabama at Birmingham, Birmingham, Alabama, USA.

4Department of Neurological Surgery, Brain Research Institute, Niigata University, Niigata, Japan.

5Department of Molecular Biosciences,

6High Throughput Analysis Laboratory, and

7Department of Neurological Surgery, Northwestern University Feinberg School of Medicine, Chicago, Illinois, USA.

8Department of Neurological Surgery, Wakayama Medical University, Wakayama, Japan.

9Institute for Cancer Genetics,

10Department of Pediatrics, and

11Department of Genetics and Development, Columbia University Irving Medical Center, New York, New York, USA.

12Center for Vascular and Developmental Biology, Feinberg Cardiovascular and Renal Research Institute, and

13Simpson Querrey Institute for BioNanotechnology, Northwestern University Feinberg School of Medicine, Chicago, Illinois, USA.

14Department of Pediatrics, Icahn School of Medicine at Mount Sinai, New York, New York, USA.

15Division of Pediatric Hematology and Oncology, Children’s of Alabama, Birmingham, Alabama, USA.

16O’Neal Comprehensive Cancer Center, University of Alabama at Birmingham, Birmingham, Alabama, USA.

Address correspondence to: Oren J. Becher, Department of Pediatrics, Icahn School of Medicine at Mount Sinai, 1468 Madison Avenue, 4th Floor, New York, New York 10029, USA. Phone: 212.241.7022; Email: oren.becher@mssm.edu. Or to: Rintaro Hashizume, Department of Pediatrics, University of Alabama at Birmingham, 1670 University Boulevard, G019A, Birmingham, Alabama 35294, USA. Phone: 205.975.0285; Email: rhashi23@uab.edu.

Find articles by Zhang, Z. in: JCI | PubMed | Google Scholar

1Department of Pediatrics, Northwestern University Feinberg School of Medicine, Chicago, Illinois, USA.

2Division of Hematology, Oncology, Neuro-Oncology and Stem Cell Transplantation, Ann & Robert H. Lurie Children’s Hospital of Chicago, Chicago, Illinois, USA.

3Department of Pediatrics, University of Alabama at Birmingham, Birmingham, Alabama, USA.

4Department of Neurological Surgery, Brain Research Institute, Niigata University, Niigata, Japan.

5Department of Molecular Biosciences,

6High Throughput Analysis Laboratory, and

7Department of Neurological Surgery, Northwestern University Feinberg School of Medicine, Chicago, Illinois, USA.

8Department of Neurological Surgery, Wakayama Medical University, Wakayama, Japan.

9Institute for Cancer Genetics,

10Department of Pediatrics, and

11Department of Genetics and Development, Columbia University Irving Medical Center, New York, New York, USA.

12Center for Vascular and Developmental Biology, Feinberg Cardiovascular and Renal Research Institute, and

13Simpson Querrey Institute for BioNanotechnology, Northwestern University Feinberg School of Medicine, Chicago, Illinois, USA.

14Department of Pediatrics, Icahn School of Medicine at Mount Sinai, New York, New York, USA.

15Division of Pediatric Hematology and Oncology, Children’s of Alabama, Birmingham, Alabama, USA.

16O’Neal Comprehensive Cancer Center, University of Alabama at Birmingham, Birmingham, Alabama, USA.

Address correspondence to: Oren J. Becher, Department of Pediatrics, Icahn School of Medicine at Mount Sinai, 1468 Madison Avenue, 4th Floor, New York, New York 10029, USA. Phone: 212.241.7022; Email: oren.becher@mssm.edu. Or to: Rintaro Hashizume, Department of Pediatrics, University of Alabama at Birmingham, 1670 University Boulevard, G019A, Birmingham, Alabama 35294, USA. Phone: 205.975.0285; Email: rhashi23@uab.edu.

Find articles by Becher, O. in: JCI | PubMed | Google Scholar

1Department of Pediatrics, Northwestern University Feinberg School of Medicine, Chicago, Illinois, USA.

2Division of Hematology, Oncology, Neuro-Oncology and Stem Cell Transplantation, Ann & Robert H. Lurie Children’s Hospital of Chicago, Chicago, Illinois, USA.

3Department of Pediatrics, University of Alabama at Birmingham, Birmingham, Alabama, USA.

4Department of Neurological Surgery, Brain Research Institute, Niigata University, Niigata, Japan.

5Department of Molecular Biosciences,

6High Throughput Analysis Laboratory, and

7Department of Neurological Surgery, Northwestern University Feinberg School of Medicine, Chicago, Illinois, USA.

8Department of Neurological Surgery, Wakayama Medical University, Wakayama, Japan.

9Institute for Cancer Genetics,

10Department of Pediatrics, and

11Department of Genetics and Development, Columbia University Irving Medical Center, New York, New York, USA.

12Center for Vascular and Developmental Biology, Feinberg Cardiovascular and Renal Research Institute, and

13Simpson Querrey Institute for BioNanotechnology, Northwestern University Feinberg School of Medicine, Chicago, Illinois, USA.

14Department of Pediatrics, Icahn School of Medicine at Mount Sinai, New York, New York, USA.

15Division of Pediatric Hematology and Oncology, Children’s of Alabama, Birmingham, Alabama, USA.

16O’Neal Comprehensive Cancer Center, University of Alabama at Birmingham, Birmingham, Alabama, USA.

Address correspondence to: Oren J. Becher, Department of Pediatrics, Icahn School of Medicine at Mount Sinai, 1468 Madison Avenue, 4th Floor, New York, New York 10029, USA. Phone: 212.241.7022; Email: oren.becher@mssm.edu. Or to: Rintaro Hashizume, Department of Pediatrics, University of Alabama at Birmingham, 1670 University Boulevard, G019A, Birmingham, Alabama 35294, USA. Phone: 205.975.0285; Email: rhashi23@uab.edu.

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Published May 21, 2024 - More info

Published in Volume 134, Issue 13 on July 1, 2024
J Clin Invest. 2024;134(13):e174794. https://doi.org/10.1172/JCI174794.
© 2024 Watanabe 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 May 21, 2024 - Version history
Received: August 17, 2023; Accepted: May 15, 2024 View PDF Abstract

Diffuse midline glioma (DMG) H3K27-altered is one of the most malignant childhood cancers. Radiation therapy remains the only effective treatment yet provides a 5-year survival rate of only 1%. Several clinical trials have attempted to enhance radiation antitumor activity using radiosensitizing agents, although none have been successful. Given this, there is a critical need for identifying effective therapeutics to enhance radiation sensitivity for the treatment of DMG. Using high-throughput radiosensitivity screening, we identified bromo- and extraterminal domain (BET) protein inhibitors as potent radiosensitizers in DMG cells. Genetic and pharmacologic inhibition of BET bromodomain activity reduced DMG cell proliferation and enhanced radiation-induced DNA damage by inhibiting DNA repair pathways. RNA-Seq and the CUT&RUN (cleavage under targets and release using nuclease) analysis showed that BET bromodomain inhibitors regulated the expression of DNA repair genes mediated by H3K27 acetylation at enhancers. BET bromodomain inhibitors enhanced DMG radiation response in patient-derived xenografts as well as genetically engineered mouse models. Together, our results highlight BET bromodomain inhibitors as potential radiosensitizer and provide a rationale for developing combination therapy with radiation for the treatment of DMG.

Graphical Abstract Introduction

Diffuse midline gliomas (DMGs) with H3K27M mutation (histone H3 lysine 27 replaced with methionine) are diffusely infiltrating glial neoplasms affecting midline structures of the CNS (1). DMG is one of the most malignant childhood tumors, with a median survival of 9–12 months from diagnosis (2). Factors that contribute to the dismal prognosis include the infiltrative nature and anatomic location of the tumor within the pons, which precludes surgical resection. The identification of effective therapies has been extremely challenging, with more than 250 clinical trials involving different combinations of chemotherapeutic agents commonly used in adult glioma proving ineffective in treating DMG (3). Fractionated focal radiation to a total dose of 54–60 Gy over a 6-week period remains the only standard treatment modality that can provide transient symptom relief and a delay in tumor progression in about 70%–80% of patients. However, radiation-treated children with DMG show evidence of disease progression within the first year of completing radiation therapy (4, 5). Given this reality, the identification of efficacious therapeutic agents that enhance the antitumor effects of radiation is urgently needed for improving treatment outcomes for this patient population.

In contrast to adult gliomas, DMG is uniquely dependent on the H3K27M mutation for its initiation and maintenance (6–9). H3K27M mutation occurs in H3F3A and HIST1H3B/C genes, encoding histone H3 variants H3.3 and H3.1, respectively, in as much as 80% of DMGs and is associated with shorter survival among patients with DMG (6, 7, 10). We and others have identified a key functional consequence of H3K27M mutation: mutant protein sequestration of the polycomb repressive complex 2 (PRC2) methyltransferase resulting in functional inactivation of PRC2 (8, 9, 11, 12). This inactivation leads to a global reduction of H3K27 di-methylation (K27me2) and tri-methylation (K27me3), which, in turn, leads to extensive transcriptional reprogramming of mutant cells and promotes a stem cell–like, therapy-resistant phenotype.

While decreasing H3K27 methylation, K27M mutation also increases K27 acetylation (K27ac), which is necessary for bromo- and extraterminal domain (BET) protein transcriptional activation through RNA polymerase II (13–15). We have previously profiled the epigenome of H3K27M-mutant DMG cells and found that K27M mutant colocalized with K27ac (15). Heterotypic H3K27M-K27ac nucleosomes colocalized with BET bromodomain proteins 2 and 4 (BRD2 and BRD4) at actively transcribed gene loci to activate transcription in DMG (15). Highly selective BET bromodomain inhibitors, such as JQ1 and I-BET (16–19), suppress gene transcription by blocking binding between BRD and acetylated histones, representing a promising therapeutic strategy for treating DMG (13–15, 20). Indeed, targeted BET bromodomain activity using JQ1 inhibited tumor growth and extended the survival of animals bearing H3K27M-mutant DMG patient–derived xenografts (PDXs) (15). Because of their promising antitumor activity, BET bromodomain inhibitors are being tested in several clinical trials with cancer patients including those with myeloma (ClinicalTrials.gov NCT03068351), leukemia (NCT02158858), prostate cancer (NCT02711956), and other advanced solid cancers (NCT01587703, NCT02419417), including pediatric brain cancer (NCT03936465). However, how small molecule inhibitors, such as those targeting BET bromodomain activity, interact with radiation has not to our knowledge been explored in DMG.

Through an unbiased high-throughput radiosensitivity screen, we found the BET bromodomain inhibitors to be potent radiosensitizers of H3K27M-mutant DMG cells. The depletion of BRD using shRNAs and sgRNAs, and BET bromodomain inhibition using small molecule inhibitors, reduced DMG cell proliferation and enhanced radiation-induced DNA damage by inhibiting DNA repair pathways. Moreover, BET bromodomain inhibition downregulated expression of DNA repair genes associated with H3K27 acyetylation (H3K27ac) occupancy and enhanced DMG radiation response in vitro and in vivo. Together, these results highlight BET bromodomain inhibition as a potential radiosensitizer and provide a rationale for developing combination therapy with radiation for the treatment of this deadly pediatric brain cancer.

Results

BET bromodomain inhibitors are identified as potent radiosensitizers by high-throughput drug screening. We first performed an unbiased high-throughput radiosensitivity screen in H3.3 WT and K27M-mutant DMG neurosphere cells using a total of 2,880 compounds, including 1,280 FDA-approved drugs and 1,600 clinical compounds (mainly small molecule inhibitors of epigenetic processes) in the presence or absence of 10 Gy irradiation. Radiosensitizing effects were quantified by cell death number using confocal image analysis combined with Hoechst nuclear staining and propidium iodide (PI) DNA staining (Figure 1A). We identified several clinical-grade BET bromodomain inhibitors as potent radiosensitizers in the screen (Supplemental Table 1; supplemental material available online with this article; https://doi.org/10.1172/JCI174794DS1), which increased cell death in combination with radiation (Figure 1B and Supplemental Figure 1). H3.3 K27M-mutant DMG neurosphere cells were more sensitive to BET bromodomain inhibitors in combination with radiation than H3.3 WT DMG neurosphere cells (Supplemental Figure 1). These BET bromodomain inhibitors were subsequently validated for their antiproliferative effects in DMG cells. AZD5153 and molibresib (I-BET762) in combination with radiation showed strong additive cytotoxic effects relative to each monotherapy (white dotted line, Figure 1C). However, methotrexate and temozolomide, which have been using in combination with radiation in adult glioblastoma (GBM), did not show additive radiosensitizing effects or monotherapy cytotoxic effects in DMG cells. Our results are consistent with the results from clinical trials, which show that DMG transiently responds to the combination of temozolomide and radiation, but with no survival benefit from the combination therapy (21).

Figure 1

High-throughput drug screening with radiation identified BET bromodomain inhibitors as radiosensitizers in DMG cells. Tumor cells isolated from GEMM-D