Abstract
The purpose of this study was to compare the expression and function of NOTCH1 in chronic lymphocytic leukemia (CLL) patients harboring a wild-type (WT) or mutated NOTCH1 gene. NOTCH1 mRNA and surface protein expression levels were independent of the NOTCH1 gene mutational status, consistent with the requirement for NOTCH1 signaling in this leukemia. However, compared with NOTCH1-WT CLL, mutated cases displayed biochemical and transcriptional evidence of an intense activation of the NOTCH1 pathway. In vivo, expression and activation of NOTCH1 was highest in CLL cells from the lymph nodes as confirmed by immunohistochemistry. In vitro, the NOTCH1 pathway was rapidly downregulated, suggesting that signaling relies upon micro-environmental interactions even in NOTCH1-mutated cases. Accordingly, co-culture of Jagged1+ (the NOTCH1 ligand) nurse-like cells with autologous CLL cells sustained NOTCH1 activity over time and mediated CLL survival and resistance against pro-apoptotic stimuli, both abrogated when NOTCH1 signaling was pharmacologically switched off. Together, these results show that NOTCH1 mutations have stabilizing effects on the NOTCH1 pathway in CLL. Furthermore, micro-environmental interactions appear critical in activating the NOTCH1 pathway both in WT and mutated patients. Finally, NOTCH1 signals may create conditions that favor drug resistance, thus making NOTCH1 a potential molecular target in CLL.
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Acknowledgements
Work supported by grants from the Italian Ministries of Education, University and Research (Futuro in Ricerca 2008 no. RBFR08ATLH and 2012 no. RBFRI2DICB), Italian Ministry of Health (Bando Giovani Ricercatori 2008 no. GR-2008-1138053 and 2010 no. GR-2010-2317594), Associazione Italiana per la Ricerca sul Cancro Foundation (IG 12754, Special Program Molecular Clinical Oncology 5 × 1000 No. 10007 and My First AIRC grant no. 13470), Compagnia di San Paolo (grant no. PMN_call_2012_0071), Fondazione Cariplo (call 2012) and local funds of the University of Turin. FA is supported by a Fondazione Veronesi fellowship. BG is supported by grant from the Croatian Ministry of Science, Education and Sport (no.198-1980955-0953). We would like to thank Katiuscia Gizzi and Maria Lamusta for their excellent technical assistance.
Author contributions
FA, BG, SS, TV and CC acquired data; DR, MC, LL GD’A and OJ provided patient samples; GI provided LN samples; FA, DR, GG and GI contributed to interpreting results; FA and SD conceived and designed the work and interpreted results; and SD wrote the paper.
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Arruga, F., Gizdic, B., Serra, S. et al. Functional impact of NOTCH1 mutations in chronic lymphocytic leukemia. Leukemia 28, 1060–1070 (2014). https://doi.org/10.1038/leu.2013.319
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DOI: https://doi.org/10.1038/leu.2013.319
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