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Acute myeloid leukemia

Inhibition of ubiquitin-specific protease 7 sensitizes acute myeloid leukemia to chemotherapy

Leukemia volume 35, pages 417–432 (2021)Cite this article

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Abstract

Resistance of acute myeloid leukemia (AML) to therapeutic agents is frequent. Consequently, the mechanisms leading to this resistance must be understood and addressed. In this paper, we demonstrate that inhibition of deubiquitinylase USP7 significantly reduces cell proliferation in vitro and in vivo, blocks DNA replication progression and increases cell death in AML. Transcriptomic dataset analyses reveal that a USP7 gene signature is highly enriched in cells from AML patients at relapse, as well as in residual blasts from patient-derived xenograft (PDX) models treated with clinically relevant doses of cytarabine, which indicates a relationship between USP7 expression and resistance to therapy. Accordingly, single-cell analysis of AML patient samples at relapse versus at diagnosis showed that a gene signature of the pre-existing subpopulation responsible for relapse is enriched in transcriptomes of patients with a high USP7 level. Furthermore, we found that USP7 interacts and modulates CHK1 protein levels and functions in AML. Finally, we demonstrated that USP7 inhibition acts in synergy with cytarabine to kill AML cell lines and primary cells of patients with high USP7 levels. Altogether, these data demonstrate that USP7 is both a marker of resistance to chemotherapy and a potential therapeutic target in overcoming resistance to treatment.

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Fig. 1: USP7 Inhibition impacts leukemic cell proliferation and viability in vitro and in vivo.
Fig. 2: USP7 inhibition impacts primary leukemic cell proliferation and viability in vitro and in vivo, without impacting normal cells.
Fig. 3: USP7 gene expression and transcriptomic gene set.
Fig. 4: USP7 and CHK1 protein expression correlate in primary AML samples.
Fig. 5: USP7 interacts and deubiquitinates CHK1 in leukemic cells.
Fig. 6: USP7 inhibition potentiates cytarabine treatment in AML cells.

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References

  1. Döhner H, Weisdorf DJ, Bloomfield CD. Acute myeloid leukemia. N Engl J Med. 2015;373:1136–52.

    PubMed  Google Scholar 

  2. Papaemmanuil E, Gerstung M, Bullinger L, Gaidzik VI, Paschka P, Roberts ND, et al. Genomic classification and prognosis in acute myeloid leukemia. N Engl J Med. 2016;374:2209–21.

    CAS  PubMed  PubMed Central  Google Scholar 

  3. Yang M, Zhao J, Liu T, Yang X, Wei H, Xu W, et al. Use of FLT3 inhibitors in acute myeloid leukemia remission induction or salvage therapy: systematic review and meta-analysis. Cancer Manag Res. 2018;10:2635–52.

    CAS  PubMed  PubMed Central  Google Scholar 

  4. DiNardo CD, Stein EM, de Botton S, Roboz GJ, Altman JK, Mims AS, et al. Durable remissions with ivosidenib in IDH1-mutated relapsed or refractory AML. N Engl J Med. 2018;378:2386–98.

    CAS  PubMed  Google Scholar 

  5. Zhou J, Chng WJ. Resistance to FLT3 inhibitors in acute myeloid leukemia: molecular mechanisms and resensitizing strategies. World J Clin Oncol. 2018;9:90–7.

    PubMed  PubMed Central  Google Scholar 

  6. Harrigan JA, Jacq X, Martin NM, Jackson SP. Deubiquitylating enzymes and drug discovery: emerging opportunities. Nat Rev Drug Discov. 2018;17:57–78.

    CAS  PubMed  Google Scholar 

  7. Basu B, Saha G, Choudhury SG, Ghosh MK. Cellular homeostasis or tumorigenesis: USP7 playing the double agent. Cancer Cell Microenviron. 2017;4:e1624.

    Google Scholar 

  8. Li M, Brooks CL, Kon N, Gu W. A dynamic role of HAUSP in the p53-Mdm2 pathway. Mol Cell. 2004;13:879–86.

    CAS  PubMed  Google Scholar 

  9. Bhattacharya S, Chakraborty D, Basu M, Ghosh MK. Emerging insights into HAUSP (USP7) in physiology, cancer and other diseases. Signal Transduct Target Ther. 2018;3:17.

    PubMed  PubMed Central  Google Scholar 

  10. Song MS, Salmena L, Carracedo A, Egia A, Lo-Coco F, Teruya-Feldstein J, et al. The deubiquitinylation and localization of PTEN are regulated by a HAUSP-PML network. Nature. 2008;455:813–7.

    CAS  PubMed  PubMed Central  Google Scholar 

  11. van der Horst A, de Vries-Smits AM, Brenkman AB, van Triest MH, van den Broek N, Colland F, et al. FOXO4 transcriptional activity is regulated by monoubiquitination and USP7/HAUSP. Nat Cell Biol. 2006;8:1064–73.

    PubMed  Google Scholar 

  12. Morotti A, Panuzzo C, Crivellaro S, Pergolizzi B, Familiari U, Berger AH, et al. BCR-ABL disrupts PTEN nuclear-cytoplasmic shuttling through phosphorylation-dependent activation of HAUSP. Leukemia. 2014;28:1326–33.

    CAS  PubMed  Google Scholar 

  13. D’Arcy P, Wang X, Linder S. Deubiquitinase inhibition as a cancer therapeutic strategy. Pharm Ther. 2015;147:32–54.

    Google Scholar 

  14. Kategaya L, Di Lello P, Rougé L, Pastor R, Clark KR, Drummond J, et al. USP7 small-molecule inhibitors interfere with ubiquitin binding. Nature. 2017;550:534–8.

    CAS  PubMed  Google Scholar 

  15. Turnbull AP, Ioannidis S, Krajewski WW, Pinto-Fernandez A, Heride C, Martin ACL, et al. Molecular basis of USP7 inhibition by selective small-molecule inhibitors. Nature. 2017;550:481–6.

    CAS  PubMed  PubMed Central  Google Scholar 

  16. Lamberto I, Liu X, Seo HS, Schauer NJ, Iacob RE, Hu W, et al. Structure-guided development of a potent and selective non-covalent active-site inhibitor of USP7. Cell Chem Biol. 2017;24:1490–500.

    CAS  PubMed  PubMed Central  Google Scholar 

  17. Pozhidaeva A, Valles G, Wang F, Wu J, Sterner DE, Nguyen P, et al. USP7-specific inhibitors target and modify the enzyme’s active site via distinct chemical mechanisms. Cell Chem Biol. 2017;24:1501–12.

    CAS  PubMed  Google Scholar 

  18. Desroses M, Altun M. The next step forward in ubiquitin-specific protease 7 selective inhibition. Cell Chem Biol. 2017;24:1429–31.

    CAS  PubMed  Google Scholar 

  19. Gavory G, O’Dowd CR, Helm MD, Flasz J, Arkoudis E, Dossang A, et al. Discovery and characterization of highly potent and selective allosteric USP7 inhibitors. Nat Chem Biol. 2018;14:118–25.

    CAS  PubMed  Google Scholar 

  20. Carra G, Panuzzo C, Torti D, Parvis G, Crivellaro S, Familiari U, et al. Therapeutic Inhibition of USP7-PTEN network in chronic lymphocytic leukemia: a strategy to overcoming TP53 mutated/deleted clones. Oncotarget. 2017;8:35508–22.

    PubMed  PubMed Central  Google Scholar 

  21. Chauhan D, Tian Z, Nicholson B, Kumar KG, Zhou B, Carrasco R, et al. A small molecule inhibitor of ubiquitin-specific protease-7 induces apoptosis in multiple myeloma cells and overcomes bortezomib resistance. Cancer Cell. 2012;22:345–58.

    CAS  PubMed  PubMed Central  Google Scholar 

  22. Fan YH, Cheng J, Vasudevan SA, Dou J, Zhang H, Patel RH, et al. USP7 inhibitor P22077 inhibits neuroblastoma growth via inducing p53-mediated apoptosis. Cell Death Dis. 2013;4:e867.

    CAS  PubMed  PubMed Central  Google Scholar 

  23. Adam K, Cartel M, Lambert M, David L, Yuan L, Besson A et al. A PIM-CHK1 signaling pathway regulates PLK1 phosphorylation and function during mitosis. J Cell Sci. 2018;10:131.

  24. David L, Fernandez-Vidal A, Bertoli S, Grgurevic S, Lepage B, Deshaies D, et al. CHK1 as a therapeutic target to bypass chemoresistance in AML. Sci Signal. 2016;9:ra90.

    PubMed  Google Scholar 

  25. Network TCGAR. Genomic and epigenomic landscapes of adult de novo acute myeloid leukemia. N Engl J Med. 2013;368:2059–74.

    Google Scholar 

  26. Verhaak RG, Wouters BJ, Erpelinck CA, Abbas S, Beverloo HB, Lugthart S, et al. Prediction of molecular subtypes in acute myeloid leukemia based on gene expression profiling. Haematologica. 2009;94:131–4.

    PubMed  Google Scholar 

  27. Hackl H, Steinleitner K, Lind K, Hofer S, Tosic N, Pavlovic S, et al. A gene expression profile associated with relapse of cytogenetically normal acute myeloid leukemia is enriched for leukemia stem cell genes. Leuk Lymphoma. 2015;56:1126–8.

    PubMed  Google Scholar 

  28. Farge T, Saland E, de Toni F, Aroua N, Hosseini M, Perry R, et al. Chemotherapy-resistant human acute myeloid leukemia cells are not enriched for leukemic stem cells but require oxidative metabolism. Cancer Discov. 2017;7:716–35.

    CAS  PubMed  PubMed Central  Google Scholar 

  29. Eppert K, Takenaka K, Lechman ER, Waldron L, Nilsson B, van Galen P, et al. Stem cell gene expression programs influence clinical outcome in human leukemia. Nat Med. 2011;17:1086–93.

    CAS  PubMed  Google Scholar 

  30. Alonsa de Vega I, Martin Y, Smits VAJ. USP7 controls Chk1 protein stability by direct deubiquitination. Cell Cycle. 2014;13:3921–6.

    Google Scholar 

  31. Zhang P, Wei Y, Wang L, Debeb BG, Yuan Y, Zhang J, et al. ATM-mediated stabilization of ZEB1 promotes DNA damage response and radioresistance through CHK1. Nat Cell Biol. 2014;16:864–75.

    CAS  PubMed  PubMed Central  Google Scholar 

  32. An T, Gong Y, Li X, Kong L, Ma P, Gong L, et al. USP7 inhibitor P5091 inhibits wnt signaling and colorectal tumor growth. Biochem Pharmacol. 2017;131:29–39.

    CAS  PubMed  Google Scholar 

  33. Petermann E, Maya-Mendoza A, Zachos G, Gillespie DA, Jackson DA, Caldecott KW. CHK1 requirement for high global rates of replication fork progression during normal vertebrate S phase. Mol Cell Biol. 2006;26:3319–26.

    CAS  PubMed  PubMed Central  Google Scholar 

  34. Lecona E, Rodriguez-Acebes S, Specks J, Lopez-Contreras AJ, Ruppen I, Murga M, et al. USP7 is a SUMO deubiquitinase essential for DNA replication. Nat Struct Mol Biol. 2016;23:270–7.

    CAS  PubMed  PubMed Central  Google Scholar 

  35. Zhao GY, Lin ZW, Lu CL, Gu J, Yuan YF, Xu FK, et al. USP7 overexpression predicts poor prognosis in lung squamous cell carcinoma and large cell carcinoma. Tumor Biol. 2015;36:1721–9.

    CAS  Google Scholar 

  36. Ma M, Yu N. Ubiquitin-specific protease 7 expression is a prognostic factor in epithelial ovarian cancer and correlates with lymph node metastasis. Onco Targets Ther. 2016;9:1559–69.

    CAS  PubMed  PubMed Central  Google Scholar 

  37. Shan H, Li X, Xiao X, Dai Y, Huang J, Song J, et al. USP7 deubiquitinates and stabilizes NOTCH1 in T-cell acute lymphoblastic leukemia. Signal Transduct Target Ther. 2018;3:29.

    PubMed  PubMed Central  Google Scholar 

  38. Jin Q, Martinez CA, Arcipowski KM, Zhu Y, Gutierrez-Diaz BT, Wang KK, et al. USP7 cooperates with NOTCH1 to drive the oncogenic transcriptional program in T cell leukemia. Clin Cancer Res. 2018;25:222–39.

    PubMed  PubMed Central  Google Scholar 

  39. Lobry C, Oh P, Mansour MR, Look AT, Aifantis I. Notch signaling: switching an oncogene to a tumor suppressor. Blood. 2014;123:2451–9.

    CAS  PubMed  PubMed Central  Google Scholar 

  40. Wang S, Itoh M, Shiratori E, Ohtaka M, Tohda S. NOTCH activation promotes glycosyltransferase expression in human myeloid leukemia cells. Hematol Rep. 2018;10:7576.

    PubMed  PubMed Central  Google Scholar 

  41. Xu X, Zhao Y, Xu M, Dai Q, Meng W, Yang J, et al. Activation of notch signal pathway is associated with a poorer prognosis in acute myeloid leukemia. Med Oncol. 2011;28:483–9.

    CAS  Google Scholar 

  42. Takam Kamga P, Bassi G, Cassaro A, Midolo M, Di Trapani M, Gatti A, et al. Notch signaling drives bone marrow stromal cell-mediated chemoresistance in acute myeloid leukemia. Oncotarget. 2016;7:21713–27.

    PubMed  Google Scholar 

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Acknowledgements

We would like to sincerely thank Thomas Farge for his technical support during in vivo experiments and Alison Boutet for technical assistance. We thank Laetitia Ligat and Dr Marie Tosolini at the CRCT microscopy facility and Manon Farcé at the CRCT cytometry facility for their helpful discussions of the analyses and the development of macro used in the ImageJ software. We gratefully acknowledge Dr Ze’ev Ronai for the critical review of the manuscript, and sincerely thank all members of the “Cell Cycle and Cancer” team for their critical review during the entire study. The authors sincerely thank Chloé Laplagne for the technical PBMC preparation and Dr Marie Jeanne Pillaire for insightful discussions on DNA fiber spreading experiments. Maëlle Cartel is a recipient of a fellowship from the Ligue Nationale contre le Cancer. We are grateful to our healthcare professionals for their boundless investment during the COVID-19 crisis.

Author information

Author notes

  1. Laure David

    Present address: IRSD, Université de Toulouse, INSERM, INRA, ENVT, UPS, Toulouse, France

  2. These authors contributed equally: Stéphane Manenti, Christine Didier

Authors and Affiliations

  1. Cancer Research Center of Toulouse, INSERM U1037, CNRS ERL 5294, Université de Toulouse, Toulouse, France

    Maëlle Cartel, Pierre-Luc Mouchel, Mathilde Gotanègre, Laure David, Sarah Bertoli, Véronique Mansat-De Mas, Jean-Emmanuel Sarry, Stéphane Manenti & Christine Didier

  2. Ligue Nationale Contre le Cancer, équipe labellisée 2016, Toulouse, France

    Maëlle Cartel, Laure David, Sarah Bertoli, Véronique Mansat-De Mas, Stéphane Manenti & Christine Didier

  3. Ligue Nationale Contre le Cancer, équipe labellisée 2018, Toulouse, France

    Pierre-Luc Mouchel, Mathilde Gotanègre & Jean-Emmanuel Sarry

  4. Service d’hématologie, Institut Universitaire du Cancer de Toulouse-Oncopôle, Toulouse, France

    Pierre-Luc Mouchel & Sarah Bertoli

  5. Laboratoire d’hématologie, Institut Universitaire du Cancer de Toulouse-Oncopole, Toulouse, France

    Véronique Mansat-De Mas

  6. LBCMCP, Centre de Biologie Intégrative, Université de Toulouse, CNRS, UPS, 31062, Toulouse Cedex, France

    Arnaud Besson

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Contributions

MC, PLM, MG, LD, JES, SM, and CD conceived and designed the study; MC, PLM, MG, LD, and CD performed experiments; SB and VMDM. contributed clinical samples; MC, AB, JES, SM, and CD wrote the manuscript; AB, JES, SM, and CD insured administrative, technical and material support; SM and CD supervised the project.

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Correspondence to Christine Didier.

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Cartel, M., Mouchel, PL., Gotanègre, M. et al. Inhibition of ubiquitin-specific protease 7 sensitizes acute myeloid leukemia to chemotherapy. Leukemia 35, 417–432 (2021). https://doi.org/10.1038/s41375-020-0878-x

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