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ResearchIn-Press PreviewHematologyMetabolism Open Access | 10.1172/jci.insight.203777

Direct pharmacological targeting of asparagine synthetase to overcome resistance to L-asparaginase in ALL therapy

Rodney Claude,1 Sankalp Srivastava,1 Kirk A. Staschke,2 Carlos A. Mellado Fritz,3 Shaoxiong Chen,4 Lei Liu,1 Minghua Zhong,1 Harish Kothandaraman,5 Nadia A. Lanman,5 Utpal P. Davé,6 Sandeep Batra,7 Jiehao Zhou,8 Yue Fang,9 Chi Zhang,9 Reuben Kapur,1 Jing Fan,3 Ronald C. Wek,2 and Ji Zhang1

1Herman B Wells Center for Pediatric Research, Indiana University School of Medicine, Indianapolis, United States of America

2Department of Biochemistry, Molecular Biology, and Pharmacology, Indiana University School of Medicine, Indianapolis, United States of America

3Morgridge Institute for Research & Department of Medical Microbiology and I, University of Wisconsin-Madison, Madison, United States of America

4Department of Pathology, Indiana University School of Medicine, Indianapolis, United States of America

5Purdue University Institute for Cancer Research, Purdue University, West Lafayette, United States of America

6Department of Medicine, Indiana University School of Medicine, Indianapolis, United States of America

7Riley Hospital for Children, Indiana University Health, Indianapolis, United States of America

8Department of Laboratory Medicine and Pathology, Mayo Clinic Arizona, Phoenix, United States of America

9Department of Medical and Molecular Genetics, Indiana University School of Medicine, Indianapolis, United States of America

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1Herman B Wells Center for Pediatric Research, Indiana University School of Medicine, Indianapolis, United States of America

2Department of Biochemistry, Molecular Biology, and Pharmacology, Indiana University School of Medicine, Indianapolis, United States of America

3Morgridge Institute for Research & Department of Medical Microbiology and I, University of Wisconsin-Madison, Madison, United States of America

4Department of Pathology, Indiana University School of Medicine, Indianapolis, United States of America

5Purdue University Institute for Cancer Research, Purdue University, West Lafayette, United States of America

6Department of Medicine, Indiana University School of Medicine, Indianapolis, United States of America

7Riley Hospital for Children, Indiana University Health, Indianapolis, United States of America

8Department of Laboratory Medicine and Pathology, Mayo Clinic Arizona, Phoenix, United States of America

9Department of Medical and Molecular Genetics, Indiana University School of Medicine, Indianapolis, United States of America

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1Herman B Wells Center for Pediatric Research, Indiana University School of Medicine, Indianapolis, United States of America

2Department of Biochemistry, Molecular Biology, and Pharmacology, Indiana University School of Medicine, Indianapolis, United States of America

3Morgridge Institute for Research & Department of Medical Microbiology and I, University of Wisconsin-Madison, Madison, United States of America

4Department of Pathology, Indiana University School of Medicine, Indianapolis, United States of America

5Purdue University Institute for Cancer Research, Purdue University, West Lafayette, United States of America

6Department of Medicine, Indiana University School of Medicine, Indianapolis, United States of America

7Riley Hospital for Children, Indiana University Health, Indianapolis, United States of America

8Department of Laboratory Medicine and Pathology, Mayo Clinic Arizona, Phoenix, United States of America

9Department of Medical and Molecular Genetics, Indiana University School of Medicine, Indianapolis, United States of America

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1Herman B Wells Center for Pediatric Research, Indiana University School of Medicine, Indianapolis, United States of America

2Department of Biochemistry, Molecular Biology, and Pharmacology, Indiana University School of Medicine, Indianapolis, United States of America

3Morgridge Institute for Research & Department of Medical Microbiology and I, University of Wisconsin-Madison, Madison, United States of America

4Department of Pathology, Indiana University School of Medicine, Indianapolis, United States of America

5Purdue University Institute for Cancer Research, Purdue University, West Lafayette, United States of America

6Department of Medicine, Indiana University School of Medicine, Indianapolis, United States of America

7Riley Hospital for Children, Indiana University Health, Indianapolis, United States of America

8Department of Laboratory Medicine and Pathology, Mayo Clinic Arizona, Phoenix, United States of America

9Department of Medical and Molecular Genetics, Indiana University School of Medicine, Indianapolis, United States of America

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1Herman B Wells Center for Pediatric Research, Indiana University School of Medicine, Indianapolis, United States of America

2Department of Biochemistry, Molecular Biology, and Pharmacology, Indiana University School of Medicine, Indianapolis, United States of America

3Morgridge Institute for Research & Department of Medical Microbiology and I, University of Wisconsin-Madison, Madison, United States of America

4Department of Pathology, Indiana University School of Medicine, Indianapolis, United States of America

5Purdue University Institute for Cancer Research, Purdue University, West Lafayette, United States of America

6Department of Medicine, Indiana University School of Medicine, Indianapolis, United States of America

7Riley Hospital for Children, Indiana University Health, Indianapolis, United States of America

8Department of Laboratory Medicine and Pathology, Mayo Clinic Arizona, Phoenix, United States of America

9Department of Medical and Molecular Genetics, Indiana University School of Medicine, Indianapolis, United States of America

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1Herman B Wells Center for Pediatric Research, Indiana University School of Medicine, Indianapolis, United States of America

2Department of Biochemistry, Molecular Biology, and Pharmacology, Indiana University School of Medicine, Indianapolis, United States of America

3Morgridge Institute for Research & Department of Medical Microbiology and I, University of Wisconsin-Madison, Madison, United States of America

4Department of Pathology, Indiana University School of Medicine, Indianapolis, United States of America

5Purdue University Institute for Cancer Research, Purdue University, West Lafayette, United States of America

6Department of Medicine, Indiana University School of Medicine, Indianapolis, United States of America

7Riley Hospital for Children, Indiana University Health, Indianapolis, United States of America

8Department of Laboratory Medicine and Pathology, Mayo Clinic Arizona, Phoenix, United States of America

9Department of Medical and Molecular Genetics, Indiana University School of Medicine, Indianapolis, United States of America

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

1Herman B Wells Center for Pediatric Research, Indiana University School of Medicine, Indianapolis, United States of America

2Department of Biochemistry, Molecular Biology, and Pharmacology, Indiana University School of Medicine, Indianapolis, United States of America

3Morgridge Institute for Research & Department of Medical Microbiology and I, University of Wisconsin-Madison, Madison, United States of America

4Department of Pathology, Indiana University School of Medicine, Indianapolis, United States of America

5Purdue University Institute for Cancer Research, Purdue University, West Lafayette, United States of America

6Department of Medicine, Indiana University School of Medicine, Indianapolis, United States of America

7Riley Hospital for Children, Indiana University Health, Indianapolis, United States of America

8Department of Laboratory Medicine and Pathology, Mayo Clinic Arizona, Phoenix, United States of America

9Department of Medical and Molecular Genetics, Indiana University School of Medicine, Indianapolis, United States of America

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

1Herman B Wells Center for Pediatric Research, Indiana University School of Medicine, Indianapolis, United States of America

2Department of Biochemistry, Molecular Biology, and Pharmacology, Indiana University School of Medicine, Indianapolis, United States of America

3Morgridge Institute for Research & Department of Medical Microbiology and I, University of Wisconsin-Madison, Madison, United States of America

4Department of Pathology, Indiana University School of Medicine, Indianapolis, United States of America

5Purdue University Institute for Cancer Research, Purdue University, West Lafayette, United States of America

6Department of Medicine, Indiana University School of Medicine, Indianapolis, United States of America

7Riley Hospital for Children, Indiana University Health, Indianapolis, United States of America

8Department of Laboratory Medicine and Pathology, Mayo Clinic Arizona, Phoenix, United States of America

9Department of Medical and Molecular Genetics, Indiana University School of Medicine, Indianapolis, United States of America

Find articles by Kothandaraman, H. in: PubMed | Google Scholar

1Herman B Wells Center for Pediatric Research, Indiana University School of Medicine, Indianapolis, United States of America

2Department of Biochemistry, Molecular Biology, and Pharmacology, Indiana University School of Medicine, Indianapolis, United States of America

3Morgridge Institute for Research & Department of Medical Microbiology and I, University of Wisconsin-Madison, Madison, United States of America

4Department of Pathology, Indiana University School of Medicine, Indianapolis, United States of America

5Purdue University Institute for Cancer Research, Purdue University, West Lafayette, United States of America

6Department of Medicine, Indiana University School of Medicine, Indianapolis, United States of America

7Riley Hospital for Children, Indiana University Health, Indianapolis, United States of America

8Department of Laboratory Medicine and Pathology, Mayo Clinic Arizona, Phoenix, United States of America

9Department of Medical and Molecular Genetics, Indiana University School of Medicine, Indianapolis, United States of America

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1Herman B Wells Center for Pediatric Research, Indiana University School of Medicine, Indianapolis, United States of America

2Department of Biochemistry, Molecular Biology, and Pharmacology, Indiana University School of Medicine, Indianapolis, United States of America

3Morgridge Institute for Research & Department of Medical Microbiology and I, University of Wisconsin-Madison, Madison, United States of America

4Department of Pathology, Indiana University School of Medicine, Indianapolis, United States of America

5Purdue University Institute for Cancer Research, Purdue University, West Lafayette, United States of America

6Department of Medicine, Indiana University School of Medicine, Indianapolis, United States of America

7Riley Hospital for Children, Indiana University Health, Indianapolis, United States of America

8Department of Laboratory Medicine and Pathology, Mayo Clinic Arizona, Phoenix, United States of America

9Department of Medical and Molecular Genetics, Indiana University School of Medicine, Indianapolis, United States of America

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1Herman B Wells Center for Pediatric Research, Indiana University School of Medicine, Indianapolis, United States of America

2Department of Biochemistry, Molecular Biology, and Pharmacology, Indiana University School of Medicine, Indianapolis, United States of America

3Morgridge Institute for Research & Department of Medical Microbiology and I, University of Wisconsin-Madison, Madison, United States of America

4Department of Pathology, Indiana University School of Medicine, Indianapolis, United States of America

5Purdue University Institute for Cancer Research, Purdue University, West Lafayette, United States of America

6Department of Medicine, Indiana University School of Medicine, Indianapolis, United States of America

7Riley Hospital for Children, Indiana University Health, Indianapolis, United States of America

8Department of Laboratory Medicine and Pathology, Mayo Clinic Arizona, Phoenix, United States of America

9Department of Medical and Molecular Genetics, Indiana University School of Medicine, Indianapolis, United States of America

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

1Herman B Wells Center for Pediatric Research, Indiana University School of Medicine, Indianapolis, United States of America

2Department of Biochemistry, Molecular Biology, and Pharmacology, Indiana University School of Medicine, Indianapolis, United States of America

3Morgridge Institute for Research & Department of Medical Microbiology and I, University of Wisconsin-Madison, Madison, United States of America

4Department of Pathology, Indiana University School of Medicine, Indianapolis, United States of America

5Purdue University Institute for Cancer Research, Purdue University, West Lafayette, United States of America

6Department of Medicine, Indiana University School of Medicine, Indianapolis, United States of America

7Riley Hospital for Children, Indiana University Health, Indianapolis, United States of America

8Department of Laboratory Medicine and Pathology, Mayo Clinic Arizona, Phoenix, United States of America

9Department of Medical and Molecular Genetics, Indiana University School of Medicine, Indianapolis, United States of America

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

1Herman B Wells Center for Pediatric Research, Indiana University School of Medicine, Indianapolis, United States of America

2Department of Biochemistry, Molecular Biology, and Pharmacology, Indiana University School of Medicine, Indianapolis, United States of America

3Morgridge Institute for Research & Department of Medical Microbiology and I, University of Wisconsin-Madison, Madison, United States of America

4Department of Pathology, Indiana University School of Medicine, Indianapolis, United States of America

5Purdue University Institute for Cancer Research, Purdue University, West Lafayette, United States of America

6Department of Medicine, Indiana University School of Medicine, Indianapolis, United States of America

7Riley Hospital for Children, Indiana University Health, Indianapolis, United States of America

8Department of Laboratory Medicine and Pathology, Mayo Clinic Arizona, Phoenix, United States of America

9Department of Medical and Molecular Genetics, Indiana University School of Medicine, Indianapolis, United States of America

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

1Herman B Wells Center for Pediatric Research, Indiana University School of Medicine, Indianapolis, United States of America

2Department of Biochemistry, Molecular Biology, and Pharmacology, Indiana University School of Medicine, Indianapolis, United States of America

3Morgridge Institute for Research & Department of Medical Microbiology and I, University of Wisconsin-Madison, Madison, United States of America

4Department of Pathology, Indiana University School of Medicine, Indianapolis, United States of America

5Purdue University Institute for Cancer Research, Purdue University, West Lafayette, United States of America

6Department of Medicine, Indiana University School of Medicine, Indianapolis, United States of America

7Riley Hospital for Children, Indiana University Health, Indianapolis, United States of America

8Department of Laboratory Medicine and Pathology, Mayo Clinic Arizona, Phoenix, United States of America

9Department of Medical and Molecular Genetics, Indiana University School of Medicine, Indianapolis, United States of America

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

1Herman B Wells Center for Pediatric Research, Indiana University School of Medicine, Indianapolis, United States of America

2Department of Biochemistry, Molecular Biology, and Pharmacology, Indiana University School of Medicine, Indianapolis, United States of America

3Morgridge Institute for Research & Department of Medical Microbiology and I, University of Wisconsin-Madison, Madison, United States of America

4Department of Pathology, Indiana University School of Medicine, Indianapolis, United States of America

5Purdue University Institute for Cancer Research, Purdue University, West Lafayette, United States of America

6Department of Medicine, Indiana University School of Medicine, Indianapolis, United States of America

7Riley Hospital for Children, Indiana University Health, Indianapolis, United States of America

8Department of Laboratory Medicine and Pathology, Mayo Clinic Arizona, Phoenix, United States of America

9Department of Medical and Molecular Genetics, Indiana University School of Medicine, Indianapolis, United States of America

Find articles by Kapur, R. in: PubMed | Google Scholar

1Herman B Wells Center for Pediatric Research, Indiana University School of Medicine, Indianapolis, United States of America

2Department of Biochemistry, Molecular Biology, and Pharmacology, Indiana University School of Medicine, Indianapolis, United States of America

3Morgridge Institute for Research & Department of Medical Microbiology and I, University of Wisconsin-Madison, Madison, United States of America

4Department of Pathology, Indiana University School of Medicine, Indianapolis, United States of America

5Purdue University Institute for Cancer Research, Purdue University, West Lafayette, United States of America

6Department of Medicine, Indiana University School of Medicine, Indianapolis, United States of America

7Riley Hospital for Children, Indiana University Health, Indianapolis, United States of America

8Department of Laboratory Medicine and Pathology, Mayo Clinic Arizona, Phoenix, United States of America

9Department of Medical and Molecular Genetics, Indiana University School of Medicine, Indianapolis, United States of America

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

1Herman B Wells Center for Pediatric Research, Indiana University School of Medicine, Indianapolis, United States of America

2Department of Biochemistry, Molecular Biology, and Pharmacology, Indiana University School of Medicine, Indianapolis, United States of America

3Morgridge Institute for Research & Department of Medical Microbiology and I, University of Wisconsin-Madison, Madison, United States of America

4Department of Pathology, Indiana University School of Medicine, Indianapolis, United States of America

5Purdue University Institute for Cancer Research, Purdue University, West Lafayette, United States of America

6Department of Medicine, Indiana University School of Medicine, Indianapolis, United States of America

7Riley Hospital for Children, Indiana University Health, Indianapolis, United States of America

8Department of Laboratory Medicine and Pathology, Mayo Clinic Arizona, Phoenix, United States of America

9Department of Medical and Molecular Genetics, Indiana University School of Medicine, Indianapolis, United States of America

Find articles by Wek, R. in: PubMed | Google Scholar

1Herman B Wells Center for Pediatric Research, Indiana University School of Medicine, Indianapolis, United States of America

2Department of Biochemistry, Molecular Biology, and Pharmacology, Indiana University School of Medicine, Indianapolis, United States of America

3Morgridge Institute for Research & Department of Medical Microbiology and I, University of Wisconsin-Madison, Madison, United States of America

4Department of Pathology, Indiana University School of Medicine, Indianapolis, United States of America

5Purdue University Institute for Cancer Research, Purdue University, West Lafayette, United States of America

6Department of Medicine, Indiana University School of Medicine, Indianapolis, United States of America

7Riley Hospital for Children, Indiana University Health, Indianapolis, United States of America

8Department of Laboratory Medicine and Pathology, Mayo Clinic Arizona, Phoenix, United States of America

9Department of Medical and Molecular Genetics, Indiana University School of Medicine, Indianapolis, United States of America

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

Published June 4, 2026 - More info

JCI Insight. https://doi.org/10.1172/jci.insight.203777.
Copyright © 2026, Claude 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 June 4, 2026 - Version history
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Abstract

Acute lymphoblastic leukemia (ALL) is the most common pediatric cancer, arising from both B- and T-cell lineages. Current therapy exploits ALL cells’ low expression of asparagine synthetase (ASNS) by using L-asparaginase, a bacterial enzyme that depletes circulating asparagine. However, resistance can emerge through induction of ASNS, mediated in part by the amino acid stress sensor GCN2. In this study, we addressed the efficacy of L-asparaginase in combination with genetic or pharmacological inhibition of GCN2 and a novel ASNS inhibitor designated ASX-173. Using a KrasG12D-driven mouse model of T-ALL, we found that GCN2 is dispensable for leukemogenesis. However, genetic inactivation or pharmacologic inhibition of GCN2 sensitized ALL cells to asparagine depletion, correlating with impaired ASNS induction. While GCN2 targeting enhanced sensitivity to asparagine depletion, a subset of Gcn2–/– T-ALL cells retained high ASNS expression and remained resistant to L-asparaginase. Likewise, some human T-ALL cells with elevated ASNS levels were refractory to GCN2 inhibition even under asparagine-depleted conditions. When combined with L-asparaginase, ASX-173 effectively eliminated ASNS-high leukemic cells in vitro and in vivo. These findings suggest that direct targeting of ASNS provides therapeutic benefit in leukemias that express high ASNS and are resistant to GCN2 inhibition under asparagine-depleted conditions.

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