Research Papers:

Droplet digital PCR analysis of NOTCH1 gene mutations in chronic lymphocytic leukemia

Angela Minervini, Crescenzio Francesco Minervini, Luisa Anelli, Antonella Zagaria, Paola Casieri, Nicoletta Coccaro, Cosimo Cumbo, Giuseppina Tota, Luciana Impera, Paola Orsini, Claudia Brunetti, Annamaria Giordano, Giorgina Specchia and Francesco Albano _

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Oncotarget. 2016; 7:86469-86479. https://doi.org/10.18632/oncotarget.13246

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Angela Minervini1,*, Crescenzio Francesco Minervini1,*, Luisa Anelli1, Antonella Zagaria1, Paola Casieri1, Nicoletta Coccaro1, Cosimo Cumbo1, Giuseppina Tota1, Luciana Impera1, Paola Orsini1, Claudia Brunetti1, Annamaria Giordano1, Giorgina Specchia1, Francesco Albano1

1Department of Emergency and Organ Transplantation (D.E.T.O.), Hematology Section, University of Bari, Bari, 70124 Italy

*These authors have contributed equally to this work

Correspondence to:

Francesco Albano, email: [email protected]

Keywords: NOTCH1, chronic lymphocytic leukemia, droplet digital PCR, IGHV mutational status, molecular monitoring

Received: September 27, 2016     Accepted: October 28, 2016     Published: November 09, 2016


In chronic lymphocytic leukemia (CLL), NOTCH1 gene mutations (NOTCH1mut) have been associated with adverse prognostic features but the independence of these as a prognostic factor is still controversial. In our study we validated a c.7541-7542delCT NOTCH1 mutation assay based on droplet digital PCR (ddPCR); we also analyzed the NOTCH1mut allelic burden, expressed as fractional abundance (FA), in 88 CLL patients at diagnosis to assess its prognostic role and made a longitudinal ddPCR analysis in 10 cases harboring NOTCH1mut to verify the FA variation over time. Our data revealed that with the ddPCR approach the incidence of NOTCH1mut in CLL was much higher (53.4%) than expected. However, longitudinal ddPCR analysis of CLL cases showed a statistically significant reduction of the NOTCH1mut FA detected at diagnosis after treatment (median FA 11.67 % vs 0.09 %, respectively, p = 0.01); the same difference, in terms of NOTCH1mut FA, was observed in the relapsed cases compared to the NOTCH1mut allelic fraction observed in patients in complete or partial remission (median FA 4.75% vs 0.43%, respectively, p = 0.007). Our study demonstrated a much higher incidence of NOTCH1mut in CLL than has previously been reported, and showed that the NOTCH1mut allelic burden evaluation by ddPCR might identify patients in need of a closer clinical follow-up during the “watch and wait” interval and after standard chemotherapy.

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