Chronic Lymphocytic Leukemia

The γ-secretase inhibitor PF-03084014 combined with fludarabine antagonizes migration, invasion and angiogenesis in NOTCH1-mutated CLL cells

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Abstract

Targeting Notch signaling has emerged as a promising therapeutic strategy for chronic lymphocytic leukemia (CLL), especially for the poor prognostic subgroup of NOTCH1-mutated patients. Here, we report that the γ-secretase inhibitor PF-03084014 inhibits the constitutive Notch activation and induces selective apoptosis in CLL cells carrying NOTCH1 mutations. Combination of PF-03084014 with fludarabine has a synergistic antileukemic effect in primary NOTCH1-mutated CLL cells, even in the presence of the protective stroma. At transcriptional level, PF-03084014 plus fludarabine treatment induces the upregulation of the proapoptotic gene HRK and the downmodulation of MMP9, IL32 and RAC2 genes that are related to invasion and chemotaxis. PF-03084014 also overcomes fludarabine-mediated activation of nuclear factor-κB signaling. Moreover, this combination impairs angiogenesis and CXCL12-induced responses in NOTCH1-mutated CLL cells, in particular those related to tumoral migration and invasion. Importantly, all these collaborative effects are specific for NOTCH1 mutation and do not occur in unmutated cases. In conclusion, we provide evidence that Notch is a therapeutic target in CLL cases with NOTCH1-activating mutations, supporting the use of Notch pathway inhibitors in combination with chemotherapy as a promising approach for the treatment of these high-risk CLL patients.

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Acknowledgements

We thank Laura Jiménez and Sandra Cabezas for their technical support. We thank Dr Maria C Cid for providing HUVEC cells and Genomics Unit of IDIBAPS for the technical help. PF-03084014 was kindly provided by Pfizer. We are also very grateful to all individuals with CLL who have participated in this study from the CLL Spanish Consortium. This work was carried out at the Esther Koplowitz Center, Barcelona. This study was supported by research funding from Ministerio de Ciencia e Innovación (SAF 12/31242 to DC and SAF 11/29326 to PP-G), Fondo de Investigación Sanitaria (PI12/01847 to GR), Redes Temáticas de Investigación Cooperativa de Cáncer from the Instituto de Salud Carlos III (ISCIII), Spanish Ministry of Economy and Competitiveness and European Regional Development Fund (ERDF) ‘Una manera de hacer Europa’ RD12/0036/0004, RD12/0036/0036; RD12/0036/0023; RD12/0036/0067 and Generalitat de Catalunya 2009SGR967 (to DC). ML-G has a contract from Fundación Científica de la Asociación Española contra el Cáncer. AM and AM-C are recipients of predoctoral fellowships FPI from Ministerio de Ciencia e Innovación. PP-G holds a contract from Ramón y Cajal program (RYC2009-05134) and GR from Miguel Servet program (PI09/00060).

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Correspondence to D Colomer.

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