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NT157, an IGF1R-IRS1/2 inhibitor, exhibits antineoplastic effects in pre-clinical models of chronic myeloid leukemia

Summary

Chronic myeloid leukemia (CML) is successfully treated with BCR-ABL1 tyrosine kinase inhibitors, but a significant percentage of patients develop resistance. Insulin receptor substrate 1 (IRS1) has been shown to constitutively associate with BCR-ABL1, and IRS1-specific silencing leads to antineoplastic effects in CML cell lines. Here, we characterized the efficacy of NT157, a pharmacological inhibitor of IGF1R-IRS1/2, in CML cells and observed significantly reduced cell viability and proliferation, accompanied by induction of apoptosis. In human K562 cells and in murine Ba/F3 cells, engineered to express either wild-type BCR-ABL1 or the imatinib-resistant BCR-ABL1T315I mutant, NT157 inhibited BCR-ABL1, IGF1R, IRS1/2, PI3K/AKT/mTOR, and STAT3/5 signaling, increased CDKN1A, FOS and JUN tumor suppressor gene expression, and reduced MYC and BCL2 oncogenes. NT157 significantly reduced colony formation of human primary CML cells with minimal effect on normal hematopoietic cells. Exposure of primary CML cells harboring BCR-ABL1T315I to NT157 resulted in increased apoptosis, reduced cell proliferation and decreased phospho-CRKL levels. In conclusion, NT157 has antineoplastic effects on BCR-ABL1 leukemogenesis, independent of T315I mutational status.

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References

  1. Deininger MW, Goldman JM, Melo JV (2000) The molecular biology of chronic myeloid leukemia. Blood 96:3343–3356

    CAS  Article  Google Scholar 

  2. O’Hare T, Eide CA, Deininger MW (2007) Bcr-Abl kinase domain mutations, drug resistance, and the road to a cure for chronic myeloid leukemia. Blood 110:2242–2249

    Article  Google Scholar 

  3. Hughes TP, Saglio G, Quintas-Cardama A, Mauro MJ, Kim DW, Lipton JH, Bradley-Garelik MB, Ukropec J, Hochhaus A (2015) BCR-ABL1 mutation development during first-line treatment with dasatinib or imatinib for chronic myeloid leukemia in chronic phase. Leukemia 29:1832–1838

    CAS  Article  Google Scholar 

  4. Apperley JF (2015) Chronic myeloid leukaemia. Lancet 385:1447–1459

    Article  Google Scholar 

  5. Wang LM, Myers MG Jr, Sun XJ, Aaronson SA, White M, Pierce JH (1993) IRS-1: essential for insulin- and IL-4-stimulated mitogenesis in hematopoietic cells. Science 261:1591–1594

    CAS  Article  Google Scholar 

  6. Folli F, Saad MJ, Backer JM, Kahn CR (1992) Insulin stimulation of phosphatidylinositol 3-kinase activity and association with insulin receptor substrate 1 in liver and muscle of the intact rat. J Biol Chem 267:22171–22177

    CAS  Article  Google Scholar 

  7. Skolnik EY, Batzer A, Li N, Lee CH, Lowenstein E, Mohammadi M, Margolis B, Schlessinger J (1993) The function of GRB2 in linking the insulin receptor to Ras signaling pathways. Science 260:1953–1955

    CAS  Article  Google Scholar 

  8. Traina F, Carvalheira JB, Saad MJ, Costa FF, Saad ST (2003) BCR-ABL binds to IRS-1 and IRS-1 phosphorylation is inhibited by imatinib in K562 cells. FEBS Lett 535:17–22

    CAS  Article  Google Scholar 

  9. Machado-Neto JA, Favaro P, Lazarini M, Costa FF, Olalla Saad ST, Traina F (2011) Knockdown of insulin receptor substrate 1 reduces proliferation and downregulates Akt/mTOR and MAPK pathways in K562 cells. Biochim Biophys Acta 1813:1404–1411

    CAS  Article  Google Scholar 

  10. Reuveni H, Flashner-Abramson E, Steiner L, Makedonski K, Song R, Shir A, Herlyn M, Bar-Eli M, Levitzki A (2013) Therapeutic destruction of insulin receptor substrates for cancer treatment. Cancer Res 73:4383–4394

    CAS  Article  Google Scholar 

  11. Ibuki N, Ghaffari M, Reuveni H, Pandey M, Fazli L, Azuma H, Gleave ME, Levitzki A, Cox ME (2014) The tyrphostin NT157 suppresses insulin receptor substrates and augments therapeutic response of prostate cancer. Mol Cancer Ther 13:2827–2839

    CAS  Article  Google Scholar 

  12. Garofalo C, Capristo M, Mancarella C, Reunevi H, Picci P, Scotlandi K (2015) Preclinical effectiveness of selective inhibitor of IRS-1/2 NT157 in osteosarcoma cell lines. Front Endocrinol (Lausanne) 6:74

    Article  Google Scholar 

  13. Flashner-Abramson E, Klein S, Mullin G, Shoshan E, Song R, Shir A, Langut Y, Bar-Eli M, Reuveni H, Levitzki A (2016) Targeting melanoma with NT157 by blocking Stat3 and IGF1R signaling. Oncogene 35:2675–2680

    CAS  Article  Google Scholar 

  14. Sanchez-Lopez E, Flashner-Abramson E, Shalapour S, Zhong Z, Taniguchi K, Levitzki A, Karin M (2016) Targeting colorectal cancer via its microenvironment by inhibiting IGF-1 receptor-insulin receptor substrate and STAT3 signaling. Oncogene 35:2634–2644

    CAS  Article  Google Scholar 

  15. Yang Y, Chan JY, Temiz NA, Yee D (2018) Insulin receptor substrate suppression by the tyrphostin NT157 inhibits responses to insulin-like growth factor-I and insulin in breast cancer cells. Horm Cancer 9(6):371–382

    CAS  Article  Google Scholar 

  16. Vardiman JW, Melo JV, Baccarani M, Radich JP, Kvasnicka HM (2017) Chronic myeloid leukaemi, BCR-ABL1-positive. In: Swerdlow SH, Campo E, Harris NL, Jaffe ES, Pileri SA, Stein H, Thiele J (eds) WHO classification of tumors of haematopoietic and lymphoid tissues. IARC, Lyon, pp 30–36

    Google Scholar 

  17. Chou TC (2006) Theoretical basis, experimental design, and computerized simulation of synergism and antagonism in drug combination studies. Pharmacol Rev 58:621–681

    CAS  Article  Google Scholar 

  18. Mulvihill MJ, Cooke A, Rosenfeld-Franklin M, Buck E, Foreman K, Landfair D, O’Connor M, Pirritt C, Sun Y, Yao Y, Arnold LD, Gibson NW, Ji QS (2009) Discovery of OSI-906: a selective and orally efficacious dual inhibitor of the IGF-1 receptor and insulin receptor. Future Med Chem 1:1153–1171

    CAS  Article  Google Scholar 

  19. Juric D, Lacayo NJ, Ramsey MC, Racevskis J, Wiernik PH, Rowe JM, Goldstone AH, O’Dwyer PJ, Paietta E, Sikic BI (2007) Differential gene expression patterns and interaction networks in BCR-ABL-positive and -negative adult acute lymphoblastic leukemias. J Clin Oncol 25:1341–1349

    CAS  Article  Google Scholar 

  20. Fenerich BA, Fernandes JC, Rodrigues Alves APN, Coelho-Silva JL, Scopim-Ribeiro R, Scheucher PS, Eide CA, Tognon CE, Druker BJ, Rego EM, Machado-Neto JA, Traina F (2020) NT157 has antineoplastic effects and inhibits IRS1/2 and STAT3/5 in JAK2V617F-positive myeloproliferative neoplasm cells. Signal Transduction Target Ther 5:5

    Article  Google Scholar 

  21. Lakshmikuttyamma A, Pastural E, Takahashi N, Sawada K, Sheridan DP, DeCoteau JF, Geyer CR (2008) Bcr-Abl induces autocrine IGF-1 signaling. Oncogene 27:3831–3844

    CAS  Article  Google Scholar 

  22. Xie J, Chen X, Zheng J, Li C, Stacy S, Holzenberger M, Hu X, Zhang CC (2015) IGF-IR determines the fates of BCR/ABL leukemia. J Hematol Oncol 8:3

    Article  Google Scholar 

  23. Abraham SA, Hopcroft LE, Carrick E, Drotar ME, Dunn K, Williamson AJ, Korfi K, Baquero P, Park LE, Scott MT, Pellicano F, Pierce A, Copland M, Nourse C, Grimmond SM, Vetrie D, Whetton AD, Holyoake TL (2016) Dual targeting of p53 and c-MYC selectively eliminates leukaemic stem cells. Nature 534:341–346

    Article  Google Scholar 

  24. Carter BZ, Mak PY, Mu H, Zhou H, Mak DH, Schober W, Leverson JD, Zhang B, Bhatia R, Huang X, Cortes J, Kantarjian H, Konopleva M, Andreeff M (2016) Combined targeting of BCL-2 and BCR-ABL tyrosine kinase eradicates chronic myeloid leukemia stem cells. Sci Transl Med 8:355ra117

    Article  Google Scholar 

  25. Passegue E, Wagner EF, Weissman IL (2004) JunB deficiency leads to a myeloproliferative disorder arising from hematopoietic stem cells. Cell 119:431–443

    CAS  Article  Google Scholar 

  26. Yang MY, Liu TC, Chang JG, Lin PM, Lin SF (2003) JunB gene expression is inactivated by methylation in chronic myeloid leukemia. Blood 101:3205–3211

    CAS  Article  Google Scholar 

  27. Hoshino K, Quintas-Cardama A, Radich J, Dai H, Yang H, Garcia-Manero G (2009) Downregulation of JUNB mRNA expression in advanced phase chronic myelogenous leukemia. Leuk Res 33:1361–1366

    CAS  Article  Google Scholar 

  28. O’Hare T, Shakespeare WC, Zhu X, Eide CA, Rivera VM, Wang F, Adrian LT, Zhou T, Huang WS, Xu Q, Metcalf CA III, Tyner JW, Loriaux MM, Corbin AS, Wardwell S, Ning Y, Keats JA, Wang Y, Sundaramoorthi R, Thomas M, Zhou D, Snodgrass J, Commodore L, Sawyer TK, Dalgarno DC, Deininger MW, Druker BJ, Clackson T (2009) AP24534, a pan-BCR-ABL inhibitor for chronic myeloid leukemia, potently inhibits the T315I mutant and overcomes mutation-based resistance. Cancer Cell 16:401–412

    Article  Google Scholar 

  29. Cortes JE, Kantarjian H, Shah NP, Bixby D, Mauro MJ, Flinn I, O’Hare T, Hu S, Narasimhan NI, Rivera VM, Clackson T, Turner CD, Haluska FG, Druker BJ, Deininger MW, Talpaz M (2012) Ponatinib in refractory Philadelphia chromosome-positive leukemias. N Engl J Med 367:2075–2088

    CAS  Article  Google Scholar 

  30. Cortes JE, Kim DW, Pinilla-Ibarz J, le Coutre P, Paquette R, Chuah C, Nicolini FE, Apperley JF, Khoury HJ, Talpaz M, DiPersio J, DeAngelo DJ, Abruzzese E, Rea D, Baccarani M, Muller MC, Gambacorti-Passerini C, Wong S, Lustgarten S, Rivera VM, Clackson T, Turner CD, Haluska FG, Guilhot F, Deininger MW, Hochhaus A, Hughes T, Goldman JM, Shah NP, Kantarjian H (2013) A phase 2 trial of ponatinib in Philadelphia chromosome-positive leukemias. N Engl J Med 369:1783–1796

    CAS  Article  Google Scholar 

  31. Moslehi JJ, Deininger M (2015) Tyrosine kinase inhibitor-associated cardiovascular toxicity in chronic myeloid leukemia. J Clin Oncol 33:4210–4218

    CAS  Article  Google Scholar 

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Acknowledgements

The authors would like to thank Dr Nicola Conran for English revision.

Funding

This study was financed in part by São Paulo Research Foundation (FAPESP), Grants #14/06037-6, #16/01639-3, #14/50947-7, #13/08135-2; in part by the Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - Brasil (CAPES); and in part by National Counsel of Technological and Scientific Development (CNPq), Grants #460750/2014-3, #305158/2013-9.

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Authors and Affiliations

Authors

Contributions

R.S-R designed, executed and analyzed the experiments and prepared the manuscript. J.A.M-N., J.L.C-S. participated in experiments and analyzed, and prepared the manuscript. B.A.F., J.C.F. provided inputs and participated in experiments using cell lines and primary human cells. P.S.S. participated in flow cytometry experiments and data analysis. C.A.E., S.L.S.S., C.E.T., B.J.D. provided inputs and participated in the interpretation of manuscript data. P.M.C., S.T.O.S., L.C.P., L.L.F-P., B.P.S, E.M.R. contributed to recruiting patients and collecting data. F.T. supervised and participated in overall design of study, experiments and analyses. All authors reviewed and edited the manuscript.

Corresponding author

Correspondence to Fabiola Traina.

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Conflict of interest

Brian J. Druker potential competing interests- SAB: Aileron Therapeutics, ALLCRON, Cepheid, Gilead Sciences, Vivid Biosciences, Celgene & Baxalta (inactive); SAB & Stock: Aptose Biosciences, Blueprint Medicines, Beta Cat, GRAIL, Third Coast Therapeutics, CTI BioPharma (inactive); Scientific Founder & Stock: MolecularMD; Board of Directors & Stock: Amgen; Board of Directors: Burroughs Wellcome Fund, CureOne; Joint Steering Committee: Beat AML LLS; Clinical Trial Funding: Novartis, Bristol-Myers Squibb, Pfizer; Royalties from Patent 6958335 (Novartis exclusive license) and OHSU and Dana-Farber Cancer Institute (one Merck exclusive license). Renata Scopim-Ribeiro declares that she has no conflict of interest. João Agostinho Machado-Neto declares that he has no conflict of interest. Christopher A. Eide declares that he has no conflict of interest. Juan Luiz Coelho-Silva declares that he has no conflict of interest. Bruna Alves Fenerich declares that she has no conflict of interest. Jaqueline Cristina Fernandes declares that she has no conflict of interest. Priscila Santos Scheucher declares that she has no conflict of interest. Samantha L. Savage Stevens declares that she has no conflict of interest. Paula de Melo Campos declares that she has no conflict of interest. Sara T. Olalla Saad declares that she has no conflict of interest. Leonardo de Carvalho Palma declares that he has no conflict of interest. Lorena Lobo de Figueiredo-Pontes declares that she has no conflict of interest. Belinda Pinto Simões declares that she has no conflict of interest. Eduardo Magalhães Rego declares that he has no conflict of interest. Cristina E. Tognon declares that she has no conflict of interest. Fabiola Traina declares that she has no conflict of interest.

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All procedures performed in studies involving human participants were in accordance with the ethical standards of the institutional and with the 1964 Helsinki declaration and its later amendments.

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Scopim-Ribeiro, R., Machado-Neto, J.A., Eide, C.A. et al. NT157, an IGF1R-IRS1/2 inhibitor, exhibits antineoplastic effects in pre-clinical models of chronic myeloid leukemia. Invest New Drugs 39, 736–746 (2021). https://doi.org/10.1007/s10637-020-01028-8

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  • DOI: https://doi.org/10.1007/s10637-020-01028-8

Keywords

  • Insulin receptor substrate 1
  • NT157
  • BCR-ABL1
  • Leukemogenesis