The Predominant Prognostic Significance of NOTCH1 Mutation Defined by Emulsion PCR in Chronic Lymphocytic Leukemia

Abstract

Katarzyna Skórka,1 Michał Chojnacki,1 Marta Masternak,1 Agnieszka Karczmarczyk,1 Edyta Subocz,2 Ewa Wawrzyniak,3 Krzysztof Giannopoulos1,4 1Department of Experimental Hematooncology, Medical University of Lublin, Lublin, Poland; 2Department of Internal Medicine and Hematology, Military Institute of Medicine, Warsaw, Poland; 3Department of Hematology, Medical University of Lodz, Lodz, Poland; 4Department of Hematology, St. John’s Cancer Centre, Lublin, PolandCorrespondence: Krzysztof GiannopoulosDepartment of Experimental Hematooncology, Medical University of Lublin, Chodzki 1, Lublin, 20-093, PolandTel + 48 81448 6632Fax + 48 81448 6634Email [email protected]: NOTCH1mut represents a new prognostic marker in chronic lymphocytic leukaemia (CLL). The low sensitivity of the current methods may increase the risk of false-negative results, particularly in patients with low NOTCH1mut allelic burden. This study compared two methods of the NOTCH1mut assessment including droplet digital PCR (ddPCR) and amplification-refractory mutation system PCR (ARMS-PCR) untreated CLL patients.Patients and Methods: This study included 319 untreated CLL patients. Two PCR-based methods; ddPCR and ARMS-PCR were performed to assess the mutational status of NOTCH1. The Mann–Whitney, Fisher’s exact test, Kruskal–Wallis, Kaplan–Meier, Log rank tests and multivariate Cox proportional hazard regression model were used to analyze collected data.Results: We proved that ddPCR increased the detectability of the NOTCH1mut compared to ARMS-PCR in CLL (18.55% vs 6%). We showed a shorter time to first treatment (TTFT) in the NOTCH1mut group of patients compared to the NOTCH1wt defined by ddPCR (1.5 vs 33 months, p=0.01). The TTFT survival curves analysis in subgroups divided according to the mutational status of IGHV and NOTCH1 assessed by ddPCR discriminated group with the best prognosis: IGHVmutNOTCH1wt. Multivariate analysis revealed that the mutational status of IGHV represented an independent prognostic factor for TTFT, while NOTCH1mut determined by ddPCR constituted as a dependent prognostic factor for TTFT.Conclusion: The selection of the precise method of NOTCH1mut detection as ddPCR might significantly improve prognostic stratification of CLL patient. Assessment of IGHV might be relevant to more accurate discrimination of prognostic groups of CLL patients, especially in harboring NOTCH1mut irrespective of the quantity of allelic burden.Keywords: chronic lymphocytic leukemia, CLL, NOTCH1, droplet digital PCR, ddPCR, amplification-refractory mutation system PCR, ARMS-PCR, prognostic marker

Keywords

• chronic lymphocytic leukemia (cll)
• notch1
• droplet digital pcr (ddpcr)
• amplification-refractory mutation system pcr (arms-pcr)
• prognostic marker