Acute myeloid leukemia

Engineering resistance to CD33-targeted immunotherapy in normal hematopoiesis by CRISPR/Cas9-deletion of CD33 exon 2

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  1. 1.

    Walter RB, Appelbaum FR, Estey EH, Bernstein ID. Acute myeloid leukemia stem cells and CD33-targeted immunotherapy. Blood. 2012;119:6198–208.

    CAS  Article  Google Scholar 

  2. 2.

    Laszlo GS, Estey EH, Walter RB. The past and future of CD33 as therapeutic target in acute myeloid leukemia. Blood Rev. 2014;28:143–53.

    CAS  Article  Google Scholar 

  3. 3.

    Godwin CD, Gale RP, Walter RB. Gemtuzumab ozogamicin in acute myeloid leukemia. Leukemia. 2017;31:1855–68.

    CAS  Article  Google Scholar 

  4. 4.

    Walter RB. Investigational CD33-targeted therapeutics for acute myeloid leukemia. Expert Opin Investig Drugs. 2018;27:339–48.

    CAS  Article  Google Scholar 

  5. 5.

    Brinkman-Van der Linden EC, Angata T, Reynolds SA, Powell LD, Hedrick SM, Varki A. CD33/Siglec-3 binding specificity, expression pattern, and consequences of gene deletion in mice. Mol Cell Biol. 2003;23:4199–206.

    CAS  Article  Google Scholar 

  6. 6.

    Kim MY, Yu KR, Kenderian SS, Ruella M, Chen S, Shin TH, et al. Genetic inactivation of CD33 in hematopoietic stem cells to enable CAR T cell immunotherapy for acute myeloid leukemia. Cell. 2018;173:1439–53.e19.

    CAS  Article  Google Scholar 

  7. 7.

    Jiang F, Doudna JA. CRISPR-Cas9 structures and mechanisms. Annu Rev Biophys. 2017;46:505–29.

    CAS  Article  Google Scholar 

  8. 8.

    Lucas D, O’Leary HA, Ebert BL, Cowan CA, Tremblay CS. Utility of CRISPR/Cas9 systems in hematology research. Exp Hematol. 2017;54:1–3.

    CAS  Article  Google Scholar 

  9. 9.

    Gundry MC, Dever DP, Yudovich D, Bauer DE, Haas S, Wilkinson AC, et al. Technical considerations for the use of CRISPR/Cas9 in hematology research. Exp Hematol. 2017;54:4–11.

    CAS  Article  Google Scholar 

  10. 10.

    Collins PJ, Hale CM, Xu H. Edited course of biomedical research: leaping forward with CRISPR. Pharmacol Res. 2017;125(Pt B):258–65.

    CAS  Article  Google Scholar 

  11. 11.

    Laszlo GS, Harrington KH, Gudgeon CJ, Beddoe ME, Fitzgibbon MP, Ries RE, et al. Expression and functional characterization of CD33 transcript variants in human acute myeloid leukemia. Oncotarget. 2016;7:43281–94.

    Article  Google Scholar 

  12. 12.

    Humbert O, Peterson CW, Norgaard ZK, Radtke S, Kiem HP. A nonhuman primate transplantation model to evaluate hematopoietic stem cell gene editing strategies for beta-hemoglobinopathies. Mol Ther Methods Clin Dev. 2018;8:75–86.

    CAS  Article  Google Scholar 

  13. 13.

    Correnti CE, Laszlo GS, de van der Schueren WJ, Godwin CD, Bandaranayake A, Busch MA, et al. Simultaneous multiple interaction T-cell engaging (SMITE) bispecific antibodies overcome bispecific T-cell engager (BiTE) resistance via CD28 co-stimulation. Leukemia. 2018;32:1239–43.

    CAS  Article  Google Scholar 

  14. 14.

    Haworth KG, Ironside C, Norgaard ZK, Obenza WM, Adair JE, Kiem HP. In vivo murine-matured human CD3( + ) cells as a preclinical model for T cell-based immunotherapies. Mol Ther Methods Clin Dev. 2017;6:17–30.

    CAS  Article  Google Scholar 

  15. 15.

    Schneider CA, Rasband WS, Eliceiri KW. NIH Image to ImageJ: 25 years of image analysis. Nat Methods. 2012;9:671–5.

    CAS  Article  Google Scholar 

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We thank Dr. Colin E. Correnti and the Molecular Design and Therapeutics core facility team (Fred Hutchinson Cancer Research Center) for the generation of AMG 330. We also thank Helen Crawford for help preparing and formatting this manuscript, and Jerry Chen for general mouse colony maintenance. Research reported in this publication was supported by the Leukemia & Lymphoma Society (Translational Research Program, grant 6489-16). RBW is a Leukemia & Lymphoma Society Scholar in Clinical Research. H-PK is a Markey Molecular Medicine Investigator and received support as the inaugural recipient of the José Carreras/E. Donnall Thomas Endowed Chair for Cancer Research and the Fred Hutchinson Cancer Research Center Endowed Chair for Cell and Gene Therapy.

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Correspondence to Olivier Humbert.

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Conflict of interest

RBW has received laboratory research grants and/or clinical trial support from Actinium Pharmaceuticals, Inc, Amgen Inc., Amphivena Therapeutics, Inc., Covagen AG, and Seattle Genetics, Inc.; has ownership interests with Amphivena Therapeutics, Inc.; and is (or has been) a consultant to Amphivena Therapeutics, Inc., Boehringer Ingelheim Pharma GmbH & Co. KG, Covagen AG, Pfizer, Inc., and Seattle Genetics, Inc. H-PK is a consultant for Rocket Pharma and Homology Medicines. The other authors declare that they have no conflict of interest.

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Humbert, O., Laszlo, G.S., Sichel, S. et al. Engineering resistance to CD33-targeted immunotherapy in normal hematopoiesis by CRISPR/Cas9-deletion of CD33 exon 2. Leukemia 33, 762–808 (2019).

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