Page 135 - Haematologica Vol. 110 - January 2025

ARTICLE - CITE-seq analysis of HU effects on CML cells
following HU treatment and a non-optimal response to TKI in the two patients with the highest LSC-II content. Although the small sample size precludes conclusions regarding the role of LSC-II in TKI responsiveness, these findings point towards analysis of LSC-II/LSC-I ratios in a larger number of HU-treated patients to elucidate long-term effects on CML outcome.
In conclusion, the results of this study imply that short-term HU therapy enhances erythroid progenitor cell differentia- tion and alters the characteristics of CML LSC. Hence, HU status should be considered in future analyses addressing the SPC compartment in CML.
Disclosures
No conflicts of interest to disclose.
Contributions
HK and MSN designed, performed and analyzed experiments. MSN designed and performed the bioinformatic analysis with input from HK. TSB performed the SCENIC analysis. LW provided the material from the chronic myeloid leukemia patients. HK and MSN isolated mononuclear cells from pa- tients’ samples. AM, FBT, KH, PJ and LW conceived, designed and supervised the study. HK, MSN, KH, FBT and AM wrote
References
1. Hungerford DA. Chromosome studies in human leukemia. 1. Acute leukemia in children. J Natl Cancer Inst. 1961;27:983-1011.
2. Quintás-Cardama A, Cortes JE. Chronic myeloid leukemia: diagnosis and treatment. Mayo Clin Proc. 2006;81(7):973-988.
3. Cortes JE, Saglio G, Kantarjian HM, et al. Final 5-year study results of DASISION: the dasatinib versus imatinib study in treatment-naïve chronic myeloid leukemia patients trial. J Clin Oncol. 2016;34(20):2333-2340.
4. Hochhaus A, Saglio G, Hughes TP, et al. Long-term benefits and risks of frontline nilotinib vs imatinib for chronic myeloid leukemia in chronic phase: 5-year update of the randomized ENESTnd trial. Leukemia. 2016;30(5):1044-1054.
5. Hehlmann R, Heimpel H, Hasford J, et al. Randomized comparison of interferon-alpha with busulfan and hydroxyurea in chronic myelogenous leukemia. The German CML Study Group. Blood. 1994;84(12):4064-4077.
6. Wolfe HR, Rein LAM. The evolving landscape of frontline therapy in chronic phase chronic myeloid leukemia (CML). Curr Hematol Malig Rep. 2021;16(5):448-454.
7. Hochhaus A, Baccarani M, Silver RT, et al. European LeukemiaNet 2020 recommendations for treating chronic myeloid leukemia. Leukemia. 2020;34(4):966-984.
8. Kockerols CCB, Geelen I, Levin MD, et al. The use of hydroxyurea pretreatment in chronic myeloid leukemia in the current tyrosine kinase inhibitor era. Haematologica. 2022;107(8):1940-1943.
9. Kennedy BJ. Hydroxyurea therapy in chronic myelogenous leukemia. Cancer. 1972;29(4):1052-1056.
10. Cokic VP, Smith RD, Beleslin-Cokic BB, et al. Hydroxyurea induces fetal hemoglobin by the nitric oxide-dependent activation of soluble guanylyl cyclase. J Clin Invest. 2003;111(2):231-239.
H. Komic et al. the manuscript. All authors approved the manuscript prior
to submission.
Acknowledgments
The authors would like to acknowledge all patients and caregivers involved in the study. Figures were created with BioRender.com.
Funding
This work was supported by grants from the Swedish Re- search Council (2020-02783 to FBT and 2023-02193 to AM), the Swedish Cancer Society (21 1864 Pj to KH, 22 2388 Pj to FBT and 22 2128 Pj to AM), the Swedish state via the ALF agreement (ALFGBG-718421 to KH, ALFGBG-963642 to FBT and ALFGBG-724881 to AM), the Swedish Childhood Cancer Fund (PR2021-0015 to PJ), the Assar Gabrielsson Foundation and the Sahlgrenska Academy at the University of Gothenburg. The authors received research funding in the form of free reagents from BD Biosciences within the BD Multiomics Alliance.
Data-sharing statement
CITE-sequencing expression data are available online in a Zenodo repository (10.5281/zenodo.12580559). R codes are available upon request. For fastq files, please contact anna. martner@gu.se.
11. Lou TF, Singh M, Mackie A, Li W, Pace BS. Hydroxyurea generates nitric oxide in human erythroid cells: mechanisms for gamma- globin gene activation. Exp Biol Med. 2009;234(11):1374-1382.
12. Subotički T, Mitrović Ajtić O, Djikić D, et al. Nitric oxide mediation in hydroxyurea and nitric oxide metabolites’ inhibition of erythroid progenitor growth. Biomolecules. 2021;11(11):1562.
13. Ikuta T, Sellak H, Odo N, Adekile AD, Gaensler KML. Nitric oxide- cGMP Signaling stimulates erythropoiesis through multiple lineage-specific transcription factors: clinical implications and a novel target for erythropoiesis. PLoS One. 2016;11(1):e0144561.
14. Hümmer J, Kraus S, Brändle K, Lee-Thedieck C. Nitric oxide in the control of the in vitro proliferation and differentiation of human hematopoietic stem and progenitor cells. Front Cell Dev Biol. 2020;8:610369.
15. Holyoake T, Jiang X, Eaves C, Eaves A. Isolation of a highly quiescent subpopulation of primitive leukemic cells in chronic myeloid leukemia. Blood. 1999;94(6):2056-2064.
16. Vetrie D, Helgason GV, Copland M. The leukaemia stem cell: similarities, differences and clinical prospects in CML and AML. Nat Rev Cancer. 2020;20(3):158-173.
17. Houshmand M, Simonetti G, Circosta P, et al. Chronic myeloid leukemia stem cells. Leukemia. 2019;33(7):1543-1556.
18. Mojtahedi H, Yazdanpanah N, Rezaei N. Chronic myeloid leukemia stem cells: targeting therapeutic implications. Stem Cell Res Ther. 2021;12(1):603.
19. Nilsson MS, Komic H, Gustafsson J, et al. Multiomic single-cell analysis identifies von Willebrand factor and TIM3-expressing BCR-ABL1+ CML stem cells. bioRxiv. 2023 Sep 17. doi:10.1101/2023.09.14.557507 [preprint, not peer-reviewed].
20. Hao Y, Hao S, Andersen-Nissen E, et al. Integrated analysis of
Haematologica | 110 January 2025
127