Page 127 - Haematologica June

Non-Hodgkin Lymphoma
Highly sensitive MYD88L265P mutation detection by droplet digital polymerase chain reaction
in Waldenström macroglobulinemia
Ferrata Storti Foundation
Daniela Drandi,1 Elisa Genuardi,1 Irene Dogliotti,1,2 Martina Ferrante,1
Cristina Jiménez,3 Francesca Guerrini,4 Mariella Lo Schirico,1
Barbara Mantoan,1 Vittorio Muccio,2 Giuseppe Lia,1 Gian Maria Zaccaria,1,5 Paola Omedè,2 Roberto Passera,6 Lorella Orsucci,7 Giulia Benevolo,7
Federica Cavallo,1,2 Sara Galimberti,4 Ramón García Sanz,3 Mario Boccadoro,1,2 Marco Ladetto8 and Simone Ferrero1,2
1Department of Molecular Biotechnologies and Health Sciences, Hematology Division, University of Torino, Italy; 2Division of Hematology 1U, AOU Città della Salute e della Scienza di Torino, Italy; 3Hematology Department, University Hospital of Salamanca and Research Biomedical Institute of Salamanca, Spain; 4Division of Hematology, Department of Oncology, Santa Chiara Hospital, Pisa, Italy; 5Biolab, Department of Electronics and Telecommunications, Politecnico di Torino, Italy; 6Biostatistics Unit, Division of Nuclear Medicine, AOU Città della Salute e della Scienza di Torino, Italy; 7Division of Hematology 2, AOU Città della Salute e della Scienza di Torino, Italy and 8Division of Hematology, AO SS Antonio e Biagio e Cesare Arrigo, Alessandria, Italy
Haematologica 2018 Volume 103(6):1029-1037
ABSTRACT
We here describe a novel method for MYD88L265P mutation detec- tion and minimal residual disease monitoring in Waldenström macroglobulinemia, by droplet digital polymerase chain reac- tion, in bone marrow and peripheral blood cells, as well as in circulating cell-free DNA. Our method shows a sensitivity of 5.00x10-5, which is far superior to the widely used allele-specific polymerase chain reaction (1.00x10-3). Overall, 291 unsorted samples from 148 patients (133 with Waldenström macroglobulinemia, 11 with IgG lymphoplasmacytic lym- phoma and 4 with IgM monoclonal gammopathy of undetermined sig- nificance) were analyzed: 194 were baseline samples and 97 were follow- up samples. One hundred and twenty-two of 128 (95.3%) bone marrow and 47/66 (71.2%) baseline peripheral blood samples scored positive for MYD88L265P. To investigate whether MYD88L265P detection by droplet dig- ital polymerase chain reaction could be used for minimal residual disease monitoring, mutation levels were compared with IGH-based minimal residual disease analysis in 10 patients, and was found to be as informa- tive as the classical, standardized, but not yet validated in Waldenström macroglobulinemia, IGH-based minimal residual disease assay (r2=0.64). Finally, MYD88L265P detection by droplet digital polymerase chain reaction on plasma circulating tumor DNA from 60 patients showed a good cor- relation with bone marrow findings (bone marrow median mutational value 1.92x10-2, plasma circulating tumor DNA value: 1.4x10-2, peripheral blood value: 1.03x10-3). This study indicates that droplet digital poly- merase chain reaction assay of MYD88L265P is a feasible and sensitive tool for mutation screening and minimal residual disease monitoring in Waldenström macroglobulinemia. Both unsorted bone marrow and peripheral blood samples can be reliably tested, as can circulating tumor DNA, which represents an attractive, less invasive alternative to bone marrow for MYD88L265P detection.
Correspondence:
daniela.drandi@unito.it
Received: December 18, 2017. Accepted: February 23, 2018. Pre-published: March 22, 2018.
doi:10.3324/haematol.2017.186528
Check the online version for the most updated information on this article, online supplements, and information on authorship & disclosures: www.haematologica.org/content/103/6/1029
©2018 Ferrata Storti Foundation
Material published in Haematologica is covered by copyright. All rights are reserved to the Ferrata Storti Foundation. Use of published material is allowed under the following terms and conditions: https://creativecommons.org/licenses/by-nc/4.0/legalcode. Copies of published material are allowed for personal or inter- nal use. Sharing published material for non-commercial pur- poses is subject to the following conditions: https://creativecommons.org/licenses/by-nc/4.0/legalcode, sect. 3. Reproducing and sharing published material for com- mercial purposes is not allowed without permission in writing from the publisher.
haematologica | 2018; 103(6)
1029
ARTICLE