[en] Activating mutations of the interleukin-7 receptor (IL7R) occur in approximately 10% of patients with T cell acute lymphoblastic leukaemia (T-ALL). Most mutations generate a cysteine at the transmembrane domain leading to receptor homodimerization through disulfide bond formation and ligand-independent activation of STAT5. We hypothesized that the reducing agent N-acetylcysteine (NAC), a well-tolerated drug used widely in clinical practice to treat acetaminophen overdose, would reduce disulfide bond formation, and inhibit mutant IL7R-mediated oncogenic signalling. We found that treatment with NAC disrupted IL7R homodimerization in IL7R-mutant DND-41 cells as assessed by non-reducing Western blot, as well as in a luciferase complementation assay. NAC led to STAT5 dephosphorylation and cell apoptosis at clinically achievable concentrations in DND-41 cells, and Ba/F3 cells transformed by an IL7R-mutant construct containing a cysteine insertion. The apoptotic effects of NAC could be rescued in part by a constitutively active allele of STAT5. Despite using doses lower than those tolerated in humans, NAC treatment significantly inhibited the progression of human DND-41 cells engrafted in immunodeficient mice. Thus, targeting leukaemogenic IL7R homodimerization with NAC offers a potentially effective and feasible therapeutic strategy that warrants testing in patients with T-ALL.
Disciplines :
Hematology
Author, co-author :
Mansour, Marc R.
Reed, Casie
Eisenberg, Amy R.
Tseng, Jen-Chieh
Twizere, Jean-Claude ; Université de Liège > Agronomie, Bio-ingénierie et Chimie (AgroBioChem) > Biologie cell. et moléc.
Daakour, Sarah ; Université de Liège > Agronomie, Bio-ingénierie et Chimie (AgroBioChem) > Biologie cell. et moléc.
Ahola, T., Fellman, V., Laaksonen, R., Laitila, J., Lapatto, R., Neuvonen, P.J. & Raivio, K.O. (1999) Pharmacokinetics of intravenous N-acetylcysteine in pre-term new-born infants. European Journal of Clinical Pharmacology, 55, 645-650.
Asai, N., Iwashita, T., Matsuyama, M. & Takahashi, M. (1995) Mechanism of activation of the ret proto-oncogene by multiple endocrine neoplasia 2A mutations. Molecular and Cellular Biology, 15, 1613-1619.
Barata, J.T., Silva, A., Brandao, J.G., Nadler, L.M., Cardoso, A.A. & Boussiotis, V.A. (2004) Activation of PI3K is indispensable for interleukin 7-mediated viability, proliferation, glucose use, and growth of T cell acute lymphoblastic leukemia cells. Journal of Experimental Medicine, 200, 659-669.
Borgstrom, L., Kagedal, B. & Paulsen, O. (1986) Pharmacokinetics of N-acetylcysteine in man. European Journal of Clinical Pharmacology, 31, 217-222.
Bose, R., Kavuri, S.M., Searleman, A.C., Shen, W., Shen, D., Koboldt, D.C., Monsey, J., Goel, N., Aronson, A.B., Li, S., Ma, C.X., Ding, L., Mardis, E.R. & Ellis, M.J. (2013) Activating HER2 mutations in HER2 gene amplification negative breast cancer. Cancer Discovery, 3, 224-237.
Brown, M., Bjorksten, A., Medved, I. & McKenna, M. (2004) Pharmacokinetics of intravenous N-acetylcysteine in men at rest and during exercise. European Journal of Clinical Pharmacology, 60, 717-723.
Cartwright, T., Senussi, O. & Grady, D. (1977) Reagents which inhibit disulphide bond formation stabilize human fibroblast interferon. Journal of General Virology, 36, 323-327.
Cassonnet, P., Rolloy, C., Neveu, G., Vidalain, P.O., Chantier, T., Pellet, J., Jones, L., Muller, M., Demeret, C., Gaud, G., Vuillier, F., Lotteau, V., Tangy, F., Favre, M. & Jacob, Y. (2011) Benchmarking a luciferase complementation assay for detecting protein complexes. Nature Methods, 8, 990-992.
Chen, N., Aleksa, K., Woodland, C., Rieder, M. & Koren, G. (2007) Prevention of ifosfamide nephrotoxicity by N-acetylcysteine: clinical pharmacokinetic considerations. The Canadian Journal of Clinical Pharmacology, 14, e246-e250.
Chen, J., Reheman, A., Gushiken, F.C., Nolasco, L., Fu, X., Moake, J.L., Ni, H. & Lopez, J.A. (2011) N-acetylcysteine reduces the size and activity of von Willebrand factor in human plasma and mice. The Journal of Clinical Investigation, 121, 593-603.
Dadi, H.K. & Roifman, C.M. (1993) Activation of phosphatidylinositol-3 kinase by ligation of the interleukin-7 receptor on human thymocytes. The Journal of Clinical Investigation, 92, 1559-1563.
De Caro, L., Ghizzi, A., Costa, R., Longo, A., Ventresca, G.P. & Lodola, E. (1989) Pharmacokinetics and bioavailability of oral acetylcysteine in healthy volunteers. Arzneimittel-Forschung, 39, 382-386.
Experts in Chronic Myeloid Leukemia (2013) The price of drugs for chronic myeloid leukemia (CML) is a reflection of the unsustainable prices of cancer drugs: from the perspective of a large group of CML experts. Blood, 121, 4439-4442.
Flex, E., Petrangeli, V., Stella, L., Chiaretti, S., Hornakova, T., Knoops, L., Ariola, C., Fodale, V., Clappier, E., Paoloni, F., Martinelli, S., Fragale, A., Sanchez, M., Tavolaro, S., Messina, M., Cazzaniga, G., Camera, A., Pizzolo, G., Tornesello, A., Vignetti, M., Battistini, A., Cave, H., Gelb, B.D., Renauld, J.C., Biondi, A., Constantinescu, S.N., Foa, R. & Tartaglia, M. (2008) Somatically acquired JAK1 mutations in adult acute lymphoblastic leukemia. Journal of Experimental Medicine, 205, 751-758.
Hertzberg, L., Vendramini, E., Ganmore, I., Cazzaniga, G., Schmitz, M., Chalker, J., Shiloh, R., Iacobucci, I., Shochat, C., Zeligson, S., Cario, G., Stanulla, M., Strehl, S., Russell, L.J., Harrison, C.J., Bornhauser, B., Yoda, A., Rechavi, G., Bercovich, D., Borkhardt, A., Kempski, H., te Kronnie, G., Bourquin, J.P., Domany, E. & Izraeli, S. (2010) Down syndrome acute lymphoblastic leukemia, a highly heterogeneous disease in which aberrant expression of CRLF2 is associated with mutated JAK2: a report from the International BFM Study Group. Blood, 115, 1006-1017.
Higuchi, M., Asao, H., Tanaka, N., Oda, K., Takeshita, T. & Sugamura, K. (1997) Regulation of IL-2 signaling. Leukemia, 11, 416-417.
Hong, S.Y., Gil, H.W., Yang, J.O., Lee, E.Y., Kim, H.K., Kim, S.H., Chung, Y.H., Lee, E.M. & Hwang, S.K. (2005) Effect of high-dose intravenous N-acetylcysteine on the concentration of plasma sulfur-containing amino acids. Korean Journal of Internal Medicine, 20, 217-223.
Igaz, P., Toth, S. & Falus, A. (2001) Biological and clinical significance of the JAK-STAT pathway; lessons from knockout mice. Inflammation Research, 50, 435-441.
Jones, A.L., Jarvie, D.R., Simpson, D., Hayes, P.C. & Prescott, L.F. (1997) Pharmacokinetics of N-acetylcysteine are altered in patients with chronic liver disease. Alimentary Pharmacology & Therapeutics, 11, 787-791.
Kalman, L., Lindegren, M.L., Kobrynski, L., Vogt, R., Hannon, H., Howard, J.T. & Buckley, R. (2004) Mutations in genes required for T-cell development: IL7R, CD45, IL2RG, JAK3, RAG1, RAG2, ARTEMIS, and ADA and severe combined immunodeficiency: HuGE review. Genetics in Medicine, 6, 16-26.
Kontro, M., Kuusanmaki, H., Eldfors, S., Pemovska, T., Rajala, H., Edgren, H., Ellonen, P., Lagstrom, S., Lundan, T., Kallioniemi, O., Mustjoki, S., Porkka, K. & Heckman, C. (2014) Novel activating STAT5B mutations as drivers of T-ALL. Leukemia, 28, 1738-1742.
McElroy, C.A., Dohm, J.A. & Walsh, S.T. (2009) Structural and biophysical studies of the human IL-7/IL-7Ralpha complex. Structure, 17, 54-65.
McElroy, C.A., Holland, P.J., Zhao, P., Lim, J.M., Wells, L., Eisenstein, E. & Walsh, S.T. (2012) Structural reorganization of the interleukin-7 signaling complex. Proceedings of the National Academy of Sciences of the United States of America, 109, 2503-2508.
Metcalfe, C., Cresswell, P., Ciaccia, L., Thomas, B. & Barclay, A.N. (2011) Labile disulfide bonds are common at the leucocyte cell surface. Open Biology, 1, 110010.
Moriggl, R., Sexl, V., Kenner, L., Duntsch, C., Stangl, K., Gingras, S., Hoffmeyer, A., Bauer, A., Piekorz, R., Wang, D., Bunting, K.D., Wagner, E.F., Sonneck, K., Valent, P., Ihle, J.N. & Beug, H. (2005) Stat5 tetramer formation is associated with leukemogenesis. Cancer Cell, 7, 87-99.
Neuwelt, E.A., Pagel, M.A., Kraemer, D.F., Peterson, D.R. & Muldoon, L.L. (2004) Bone marrow chemoprotection without compromise of chemotherapy efficacy in a rat brain tumor model. Journal of Pharmacology and Experimental Therapeutics, 309, 594-599.
Nourjah, P., Ahmad, S.R., Karwoski, C. & Willy, M. (2006) Estimates of acetaminophen (Paracetomal)-associated overdoses in the United States. Pharmacoepidemiology and Drug Safety, 15, 398-405.
Olsson, B., Johansson, M., Gabrielsson, J. & Bolme, P. (1988) Pharmacokinetics and bioavailability of reduced and oxidized N-acetylcysteine. European Journal of Clinical Pharmacology, 34, 77-82.
Onishi, M., Nosaka, T., Misawa, K., Mui, A.L., Gorman, D., McMahon, M., Miyajima, A. & Kitamura, T. (1998) Identification and characterization of a constitutively active STAT5 mutant that promotes cell proliferation. Molecular and Cellular Biology, 18, 3871-3879.
Porcu, M., Kleppe, M., Gianfelici, V., Geerdens, E., De Keersmaecker, K., Tartaglia, M., Foa, R., Soulier, J., Cauwelier, B., Uyttebroeck, A., Macintyre, E., Vandenberghe, P., Asnafi, V. & Cools, J. (2012) Mutation of the receptor tyrosine phosphatase PTPRC (CD45) in T-cell acute lymphoblastic leukemia. Blood, 119, 4476-4479.
Prescott, L.F., Donovan, J.W., Jarvie, D.R. & Proudfoot, A.T. (1989) The disposition and kinetics of intravenous N-acetylcysteine in patients with paracetamol overdosage. European Journal of Clinical Pharmacology, 37, 501-506.
Sanda, T., Tyner, J.W., Gutierrez, A., Ngo, V.N., Glover, J., Chang, B.H., Yost, A., Ma, W., Fleischman, A.G., Zhou, W., Yang, Y., Kleppe, M., Ahn, Y., Tatarek, J., Kelliher, M.A., Neuberg, D.S., Levine, R.L., Moriggl, R., Muller, M., Gray, N.S., Jamieson, C.H., Weng, A.P., Staudt, L.M., Druker, B.J. & Look, A.T. (2013) TYK2-STAT1-BCL2 pathway dependence in T-cell acute lymphoblastic leukemia. Cancer Discovery, 3, 564-577.
Santoro, M., Carlomagno, F., Romano, A., Bottaro, D.P., Dathan, N.A., Grieco, M., Fusco, A., Vecchio, G., Matoskova, B., Kraus, M.H. & Di Fiore, P.P. (1995) Activation of RET as a dominant transforming gene by germline mutations of MEN2A and MEN2B. Science, 267, 381-383.
Shochat, C., Tal, N., Bandapalli, O.R., Palmi, C., Ganmore, I., te Kronnie, G., Cario, G., Cazzaniga, G., Kulozik, A.E., Stanulla, M., Schrappe, M., Biondi, A., Basso, G., Bercovich, D., Muckenthaler, M.U. & Izraeli, S. (2011) Gain-of-function mutations in interleukin-7 receptor-alpha (IL7R) in childhood acute lymphoblastic leukemias. Journal of Experimental Medicine, 208, 901-908.
Shochat, C., Tal, N., Gryshkova, V., Birger, Y., Bandapalli, O.R., Cazzaniga, G., Gershman, N., Kulozik, A.E., Biondi, A., Mansour, M.R., Twizere, J.C., Muckenthaler, M.U., Ben-Tal, N., Constantinescu, S.N., Bercovich, D. & Izraeli, S. (2014) Novel activating mutations lacking cysteine in type I cytokine receptors in acute lymphoblastic leukemia. Blood, 124, 106-110.
Silva, A., Girio, A., Cebola, I., Santos, C.I., Antunes, F. & Barata, J.T. (2011) Intracellular reactive oxygen species are essential for PI3K/Akt/mTOR-dependent IL-7-mediated viability of T-cell acute lymphoblastic leukemia cells. Leukemia, 25, 960-967.
Smilkstein, M.J., Knapp, G.L., Kulig, K.W. & Rumack, B.H. (1988) Efficacy of oral N-acetylcysteine in the treatment of acetaminophen overdose. Analysis of the national multicenter study (1976 to 1985). The New England Journal of Medicine, 319, 1557-1562.
White, K., Bruckner, J.V. & Guess, W.L. (1973) Toxicological studies of 2-mercaptoethanol. Journal of Pharmaceutical Sciences, 62, 237-241.
Yoda, A., Yoda, Y., Chiaretti, S., Bar-Natan, M., Mani, K., Rodig, S.J., West, N., Xiao, Y., Brown, J.R., Mitsiades, C., Sattler, M., Kutok, J.L., DeAngelo, D.J., Wadleigh, M., Piciocchi, A., Dal Cin, P., Bradner, J.E., Griffin, J.D., Anderson, K.C., Stone, R.M., Ritz, J., Foa, R., Aster, J.C., Frank, D.A. & Weinstock, D.M. (2010) Functional screening identifies CRLF2 in precursor B-cell acute lymphoblastic leukemia. Proceedings of the National Academy of Sciences of the United States of America, 107, 252-257.
Zenatti, P.P., Ribeiro, D., Li, W., Zuurbier, L., Silva, M.C., Paganin, M., Tritapoe, J., Hixon, J.A., Silveira, A.B., Cardoso, B.A., Sarmento, L.M., Correia, N., Toribio, M.L., Kobarg, J., Horstmann, M., Pieters, R., Brandalise, S.R., Ferrando, A.A., Meijerink, J.P., Durum, S.K., Yunes, J.A. & Barata, J.T. (2011) Oncogenic IL7R gain-of-function mutations in childhood T-cell acute lymphoblastic leukemia. Nature Genetics, 43, 932-939.
Zhang, J., Ding, L., Holmfeldt, L., Wu, G., Heatley, S.L., Payne-Turner, D., Easton, J., Chen, X., Wang, J., Rusch, M., Lu, C., Chen, S.C., Wei, L., Collins-Underwood, J.R., Ma, J., Roberts, K.G., Pounds, S.B., Ulyanov, A., Becksfort, J., Gupta, P., Huether, R., Kriwacki, R.W., Parker, M., McGoldrick, D.J., Zhao, D., Alford, D., Espy, S., Bobba, K.C., Song, G., Pei, D., Cheng, C., Roberts, S., Barbato, M.I., Campana, D., Coustan-Smith, E., Shurtleff, S.A., Raimondi, S.C., Kleppe, M., Cools, J., Shimano, K.A., Hermiston, M.L., Doulatov, S., Eppert, K., Laurenti, E., Notta, F., Dick, J.E., Basso, G., Hunger, S.P., Loh, M.L., Devidas, M., Wood, B., Winter, S., Dunsmore, K.P., Fulton, R.S., Fulton, L.L., Hong, X., Harris, C.C., Dooling, D.J., Ochoa, K., Johnson, K.J., Obenauer, J.C., Evans, W.E., Pui, C.H., Naeve, C.W., Ley, T.J., Mardis, E.R., Wilson, R.K., Downing, J.R. & Mullighan, C.G. (2012) The genetic basis of early T-cell precursor acute lymphoblastic leukaemia. Nature, 481, 157-163.