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Ferrata Storti Foundation
Haematologica 2019 Volume 104(2):288-296
Acute Myeloid Leukemia
A novel deep targeted sequencing method for minimal residual disease monitoring in acute myeloid leukemia
Esther Onecha,1,2 Maria Linares,1,2 Inmaculada Rapado,1,2,3
Yanira Ruiz-Heredia,1,2 Pilar Martinez-Sanchez,1 Teresa Cedena,1,2,3,4 Marta Pratcorona,5 Jaime Perez Oteyza,6 Pilar Herrera,7 Eva Barragan,4,8 Pau Montesinos,4,8 Jose Antonio Garcia Vela,9 Elena Magro,10
Eduardo Anguita,11 Angela Figuera,12 Rosalia Riaza,13
Pilar Martinez-Barranco,14 Beatriz Sanchez-Vega,1,2 Josep Nomdedeu,5 Miguel Gallardo,2* Joaquin Martinez-Lopez1,2,3,4* and Rosa Ayala1,2,3,4*
1Hematology Department, Hospital Universitario 12 de Octubre, Madrid; 2Hematological Malignancies Clinical Research Unit, CNIO, Madrid; 3Centro de Investigación Biomédica en Red Cáncer (CIBERONC), Madrid; 4Complutense University, Madrid; 5Hematology Department, Hospital Santa Creu i Sant Pau, Barcelona; 6Hematology Department, Hospital Universitario Sanchinarro, Madrid; 7Hematology Department, Hospital Universitario Ramon y Cajal, Madrid; 8Hematology Department, Hospital Universitario La Fe, Valencia; 9Department of Hematology, Hospital Universitario de Getafe, Madrid; 10Hematology Department, Hospital Universitario Principe de Asturias, Madrid; 11Hematology Department, Hospital Clínico San Carlos, IdISSC, UCM, Madrid; 12Hematology Department, Hospital Universitario de la Princesa, Madrid; 13Hematology Department, Hospital Universitario Severo Ochoa, Madrid and 14Hematology Department, Hospital Universitario Fundación Alcorcón, Madrid, Spain
ABSTRACT
Ahigh proportion of patients with acute myeloid leukemia who achieve minimal residual disease negative status ultimately relapse because a fraction of pathological clones remains undetected by standard methods. We designed and validated a high-throughput sequenc- ing method for minimal residual disease assessment of cell clonotypes with mutations of NPM1, IDH1/2 and/or FLT3-single nucleotide variants. For clinical validation, 106 follow-up samples from 63 patients in complete remission were studied by sequencing, evaluating the level of mutations detected at diagnosis. The predictive value of minimal residual disease sta- tus by sequencing, multiparameter flow cytometry, or quantitative poly- merase chain reaction analysis was determined by survival analysis. The sequencing method achieved a sensitivity of 10-4 for single nucleotide vari- ants and 10-5 for insertions/deletions and could be used in acute myeloid leukemia patients who carry any mutation (86% in our diagnostic data set). Sequencing–determined minimal residual disease positive status was asso- ciated with lower disease-free survival (hazard ratio 3.4, P=0.005) and lower overall survival (hazard ratio 4.2, P<0.001). Multivariate analysis showed that minimal residual disease positive status determined by sequencing was an independent factor associated with risk of death (haz- ard ratio 4.54, P=0.005) and the only independent factor conferring risk of relapse (hazard ratio 3.76, P=0.012). This sequencing-based method simpli- fies and standardizes minimal residual disease evaluation, with high appli- cability in acute myeloid leukemia. It is also an improvement upon flow cytometry- and quantitative polymerase chain reaction-based prediction of outcomes of patients with acute myeloid leukemia and could be incorpo- rated in clinical settings and clinical trials.
Introduction
Cytogenetic and molecular alterations at diagnosis and response to treatment are the most useful criteria for predicting the relative risk of relapse in acute myeloid leukemia (AML), and for guiding the choice between chemotherapy and
*MG, JM-L and RA contributed equally to this work.
Correspondence:
rosam.ayala@salud.madrid.org
Received: April 2, 2018. Accepted: August 8, 2018. Pre-published: August 9, 2018.
doi:10.3324/haematol.2018.194712
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