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Intracellular angiopoietin-1 promotes TKI-resistance via activation of JAK/STAT5 pathway in chronic myeloid leukemia

Abstract

Drug resistance from BCR-ABL tyrosine kinase inhibitors (TKIs) and other chemotherapeutics results in treatment failure and disease progression in chronic myeloid leukemia (CML). However, the mechanism is still uncertain. In this study, we investigated the role of angiopoietin-1 (ANG-1) as a potential prognostic factor for drug resistance in CML. Both intracellular and secretory ANG-1 (iANG-1 and sANG-1) were overexpressed in multidrug-resistant CML samples. The IC50 value was higher in primary CD34+ CD38 cells with more ANG-1. Silencing ANG-1significantly sensitized three TKI-resistant CML cell lines to imatinib (IM) while recombinant human ANG-1 failed to retain cell survival in vitro. This indicated the important role of iANG-1 as opposed to sANG-1 in CML drug resistance. Moreover, a similar effect was observed in xenograft mice models bearing ANG-1-silenced CML cells. Subsequently, pathway analysis and protein validation experiments showed activation of the JAK/STAT pathway and augmentation of STAT5a phosphorylation in ANG-1 restored CML cells. Upstream Src phosphorylation, which plays a crucial role in CML drug resistance, was also upregulated as a key event in iANG-1-related JAK/STAT pathway activation. In conclusion, our study elucidated a new BCR-ABL independent molecular mechanism induced by intracytoplasmic ANG-1 overexpression as a potential strategy for overcoming CML resistance.

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Fig. 1: Critical target screening in CML at the transcriptional level using bioinformatics analysis.
Fig. 2: ANGPT1 overexpression and its encoding product ANG-1 in patients with CML and TKI-resistance.
Fig. 3: Modulation of ANG-1 expression significantly affects sensitivity of CML cells to TKIs and inhibits proliferation of CML-LSCs.
Fig. 4: Silencing ANG-1 inhibits invasion and proliferation of CML cell lines in vivo and prolongs the survival time of CML-CDX mice model.
Fig. 5: Inhibition of ANG-1 increases the sensitivity of CML-LSCs to TKI in a CML-PDX mice model.
Fig. 6: Activation of JAK/STAT5 signaling pathway mediated by intracellular ANG-1 plays a crucial role in TKI-response in CML cells.
Fig. 7: SRC phosphorylation mediates activation of JAK/STAT5 pathway induced by iANG-1 overexpression.
Fig. 8: Schematic diagram of molecular mechanism of iANG-1 involved TKI-resistance in CML.

References

  1. Hochhaus A, Larson RA, Guilhot F, Radich JP, Branford S, Hughes TP, et al. Long-term outcomes of imatinib treatment for chronic myeloid leukemia. N Engl J Med. 2017;376:917–27.

    Article  CAS  Google Scholar 

  2. Bower H, Bjorkholm M, Dickman PW, Hoglund M, Lambert PC, Andersson TM. Life expectancy of patients with chronic myeloid leukemia approaches the life expectancy of the general population. J Clin Oncol. 2016;34:2851–7.

    Article  CAS  Google Scholar 

  3. Minciacchi VR, Kumar R, Krause DS. Chronic myeloid leukemia: a model disease of the past. Cells. 2021;10:117.

    Article  Google Scholar 

  4. Braun TP, Eide CA, Druker BJ. Response and resistance to BCR-ABL1-targeted therapies. Cancer Cell. 2020;37:530–42.

    Article  CAS  Google Scholar 

  5. Mitchell R, Hopcroft LEM, Baquero P, Allan EK, Hewit K, James D, et al. Targeting BCR-ABL-independent TKI resistance in chronic myeloid leukemia by mTOR and autophagy inhibition. J Natl Cancer Inst. 2018;110:467–78.

    Article  CAS  Google Scholar 

  6. Cortes JE, Kim DW, Pinilla-Ibarz J, le Coutre P, Paquette R, Chuah C, et al. A phase 2 trial of ponatinib in Philadelphia chromosome-positive leukemias. N Engl J Med. 2013;369:1783–96.

    Article  CAS  Google Scholar 

  7. Bavaro L, Martelli M, Cavo M, Soverini S. Mechanisms of disease progression and resistance to tyrosine kinase inhibitor therapy in chronic myeloid leukemia: an update. Int J Mol Sci. 2019;20:6141.

    Article  CAS  Google Scholar 

  8. Chu S, McDonald T, Lin A, Chakraborty S, Huang Q, Snyder DS, et al. Persistence of leukemia stem cells in chronic myelogenous leukemia patients in prolonged remission with imatinib treatment. Blood. 2011;118:5565–72.

    Article  CAS  Google Scholar 

  9. Hashiyama M, Iwama A, Ohshiro K, Kurozumi K, Yasunaga K, Shimizu Y, et al. Predominant expression of a receptor tyrosine kinase, TIE, in hematopoietic stem cells and B cells. Blood. 1996;87:93–101.

    Article  CAS  Google Scholar 

  10. Teufel M, Seidel H, Kochert K, Meinhardt G, Finn RS, Llovet JM, et al. Biomarkers associated with response to regorafenib in patients with hepatocellular carcinoma. Gastroenterology. 2019;156:1731–41.

    Article  CAS  Google Scholar 

  11. Fagiani E, Lorentz P, Kopfstein L, Christofori G. Angiopoietin-1 and -2 exert antagonistic functions in tumor angiogenesis, yet both induce lymphangiogenesis. Cancer Res. 2011;71:5717–27.

    Article  CAS  Google Scholar 

  12. Liu XH, Bai CG, Yuan Y, Gong DJ, Huang SD. Angiopoietin-1 targeted RNA interference suppresses angiogenesis and tumor growth of esophageal cancer. World J Gastroenterol. 2008;14:1575–81.

    Article  CAS  Google Scholar 

  13. Torimura T, Ueno T, Kin M, Harada R, Taniguchi E, Nakamura T, et al. Overexpression of angiopoietin-1 and angiopoietin-2 in hepatocellular carcinoma. J Hepatol. 2004;40:799–807.

    Article  CAS  Google Scholar 

  14. Cheng CL, Hou HA, Jhuang JY, Lin CW, Chen CY, Tang JL, et al. High bone marrow angiopoietin-1 expression is an independent poor prognostic factor for survival in patients with myelodysplastic syndromes. Br J Cancer. 2011;105:975–82.

    Article  CAS  Google Scholar 

  15. Lee CY, Tien HF, Hu CY, Chou WC, Lin LI. Marrow angiogenesis-associated factors as prognostic biomarkers in patients with acute myelogenous leukaemia. Br J Cancer. 2007;97:877–82.

    Article  CAS  Google Scholar 

  16. Schliemann C, Bieker R, Padro T, Kessler T, Hintelmann H, Buchner T, et al. Expression of angiopoietins and their receptor Tie2 in the bone marrow of patients with acute myeloid leukemia. Haematologica. 2006;91:1203–11.

    CAS  Google Scholar 

  17. Grenga I, Kwilas AR, Donahue RN, Farsaci B, Hodge JW. Inhibition of the angiopoietin/Tie2 axis induces immunogenic modulation, which sensitizes human tumor cells to immune attack. J Immunother Cancer. 2015;3:52.

    Article  Google Scholar 

  18. Atesoglu EB, Tarkun P, Mehtap O, Demirsoy ET, Atalay F, Maden M, et al. Serum angiopoietin levels are different in acute and chronic myeloid neoplasms: angiopoietins do not only regulate tumor angiogenesis. Indian J Hematol Blood Transfus. 2016;32:162–7.

    Article  Google Scholar 

  19. Ichim CV. Kinase-independent mechanisms of resistance of leukemia stem cells to tyrosine kinase inhibitors. Stem Cells Transl Med. 2014;3:405–15.

    Article  CAS  Google Scholar 

  20. Aranda-Tavio H, Recio C, Martin-Acosta P, Guerra-Rodriguez M, Brito-Casillas Y, Blanco R, et al. JKST6, a novel multikinase modulator of the BCR-ABL1/STAT5 signaling pathway that potentiates direct BCR-ABL1 inhibition and overcomes imatinib resistance in chronic myelogenous leukemia. Biomed Pharmacother. 2021;144:112330.

    Article  CAS  Google Scholar 

  21. Huang H, Bhat A, Woodnutt G, Lappe R. Targeting the ANGPT-TIE2 pathway in malignancy. Nat Rev Cancer. 2010;10:575–85.

    Article  CAS  Google Scholar 

  22. Arai F, Hirao A, Ohmura M, Sato H, Matsuoka S, Takubo K, et al. Tie2/angiopoietin-1 signaling regulates hematopoietic stem cell quiescence in the bone marrow niche. Cell 2004;118:149–61.

    Article  CAS  Google Scholar 

  23. Parikh SM. Targeting Tie2 and the host vascular response in sepsis. Sci Transl Med. 2016;8:335fs339.

    Article  Google Scholar 

  24. Tolomeo M, Meli M, Grimaudo S. STAT5 and STAT5 inhibitors in hematological malignancies. Anticancer Agents Med Chem. 2019;19:2036–46.

    Article  CAS  Google Scholar 

  25. Gleixner KV, Schneeweiss M, Eisenwort G, Berger D, Herrmann H, Blatt K, et al. Combined targeting of STAT3 and STAT5: a novel approach to overcome drug resistance in chronic myeloid leukemia. Haematologica. 2017;102:1519–29.

    Article  CAS  Google Scholar 

  26. Dong B, Liang Z, Chen Z, Li B, Zheng L, Yang J, et al. Cryptotanshinone suppresses key onco-proliferative and drug-resistant pathways of chronic myeloid leukemia by targeting STAT5 and STAT3 phosphorylation. Sci China Life Sci. 2018;61:999–1009.

    Article  CAS  Google Scholar 

  27. Pinz S, Unser S, Rascle A. Signal transducer and activator of transcription STAT5 is recruited to c-Myc super-enhancer. BMC Mol Biol. 2016;17:10.

    Article  Google Scholar 

  28. Warsch W, Grundschober E, Sexl V. Adding a new facet to STAT5 in CML: multitasking for leukemic cells. Cell Cycle. 2013;12:1813–4.

    Article  CAS  Google Scholar 

  29. Bibi S, Arslanhan MD, Langenfeld F, Jeanningros S, Cerny-Reiterer S, Hadzijusufovic E, et al. Co-operating STAT5 and AKT signaling pathways in chronic myeloid leukemia and mastocytosis: possible new targets of therapy. Haematologica. 2014;99:417–29.

    Article  CAS  Google Scholar 

  30. Juen L, Brachet-Botineau M, Parmenon C, Bourgeais J, Herault O, Gouilleux F, et al. New inhibitor targeting signal transducer and activator of transcription 5 (STAT5) signaling in myeloid leukemias. J Med Chem. 2017;60:6119–36.

    Article  CAS  Google Scholar 

  31. Jiang X, Cheng Y, Hu C, Zhang A, Ren Y, Xu X. MicroRNA-221 sensitizes chronic myeloid leukemia cells to imatinib by targeting STAT5. Leuk Lymphoma. 2019;60:1709–20.

    Article  CAS  Google Scholar 

  32. Orlova A, Wagner C, de Araujo ED, Bajusz D, Neubauer HA, Herling M, et al. Direct targeting options for STAT3 and STAT5 in cancer. Cancers. 2019;11:1930.

    Article  CAS  Google Scholar 

  33. Mirmohammadsadegh A, Hassan M, Bardenheuer W, Marini A, Gustrau A, Nambiar S, et al. STAT5 phosphorylation in malignant melanoma is important for survival and is mediated through SRC and JAK1 kinases. J Investig Dermatol. 2006;126:2272–80.

    Article  CAS  Google Scholar 

  34. Ku M, Wall M, MacKinnon RN, Walkley CR, Purton LE, Tam C, et al. Src family kinases and their role in hematological malignancies. Leuk Lymphoma. 2015;56:577–86.

    Article  CAS  Google Scholar 

  35. Okutani Y, Kitanaka A, Tanaka T, Kamano H, Ohnishi H, Kubota Y, et al. Src directly tyrosine-phosphorylates STAT5 on its activation site and is involved in erythropoietin-induced signaling pathway. Oncogene. 2001;20:6643–50.

    Article  CAS  Google Scholar 

  36. O’Hare T, Walters DK, Stoffregen EP, Sherbenou DW, Heinrich MC, Deininger MW, et al. Combined Abl inhibitor therapy for minimizing drug resistance in chronic myeloid leukemia: Src/Abl inhibitors are compatible with imatinib. Clin Cancer Res. 2005;11:6987–93.

    Article  Google Scholar 

  37. Cortes JE, Kantarjian HM, Brummendorf TH, Kim DW, Turkina AG, Shen ZX, et al. Safety and efficacy of bosutinib (SKI-606) in chronic phase Philadelphia chromosome-positive chronic myeloid leukemia patients with resistance or intolerance to imatinib. Blood. 2011;118:4567–76.

    Article  CAS  Google Scholar 

  38. Montero JC, Seoane S, Ocana A, Pandiella A. Inhibition of SRC family kinases and receptor tyrosine kinases by dasatinib: possible combinations in solid tumors. Clin Cancer Res. 2011;17:5546–52.

    Article  CAS  Google Scholar 

  39. Ma D, Liu P, Wang P, Zhou Z, Fang Q, Wang J. PKC-beta/Alox5 axis activation promotes Bcr-Abl-independent TKI-resistance in chronic myeloid leukemia. J Cell Physiol. 2021;236:6312–27.

    Article  CAS  Google Scholar 

  40. Muller J, Sperl B, Reindl W, Kiessling A, Berg T. Discovery of chromone-based inhibitors of the transcription factor STAT5. Chembiochem. 2008;9:723–7.

    Article  Google Scholar 

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Acknowledgements

This study was supported, in part, by the National Natural Science Foundation of China (Nos. 82003835, 82160704 and 82160665), Basic Research Program of Guizhou Province Technology Bureau(No. ZK[2021] General-399, No.ZK[2022]General-451 and No. ZK[2021] General-568), Science and Technology Program of Guizhou Province Health Committee(Nos. gzwkj2021-466, Nos. gzwkj2021-158 and Nos. gzwkj2021-442), National-Local Joint EngineeringResearch Center for Innovative & Generic Chemical Drug, Guizhou. High-level Innovative Talents Supporting Program (2016-4015)Guiyang City Technology Bureau Planned Project (No. 2019‐9‐14‐8). And we gratefully appreciated the support from public experimental platform of Department of Pathology of Affiliated Hospital of Guizhou Medical University.

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DM and CLZ designed the experiments; PL, YW, YMZ and YSR performed experiments; DM and CJH completed bioinformatic analyses; DM, ZZ, PL and PW analyzed data; JSW, JYZ and PHL provided specimens; DM, CLZ and LT wrote and edited the manuscript; DM and CJH provided research reagents and valuable comments; DM, CJH and PL provided funding for the study.

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Correspondence to Jishi Wang, Chengliang Zhang or Lei Tang.

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Ma, D., Liu, P., Hu, C. et al. Intracellular angiopoietin-1 promotes TKI-resistance via activation of JAK/STAT5 pathway in chronic myeloid leukemia. Oncogene 42, 124–137 (2023). https://doi.org/10.1038/s41388-022-02536-y

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