Application of a digital PCR method for WT1 to myeloid neoplasms in CR and deep ELN WT1 molecular response (< 10 copies)
Por:
Bussaglia, E, Pratcorona, M, Carricondo, M, Sansegundo, L, Rubio, MA, Monter, A, Brell, A, Badell, I, Esteve, J, Arnan, M, Talarn, C, Tormo, M, Garcia, A, Vall-Llovera, F, Ortin, X, Pedro, C, Bargay, J, Brunet, S, Sierra, J, Nomdedeu, J
Publicada:
1 abr 2020
Ahead of Print:
1 feb 2020
Resumen:
Bone marrow WT1 mRNA levels assessed by the ELN method are useful to establish prognostic correlations in myeloid malignancies treated with chemotherapy or hematopoietic stem cell transplantation (HCT). Those patients with WT1 levels below ten copies have a good outcome. However, some of these patients relapse. To further characterize this group of cases, we applied a new and sensitive digital (ddPCR) WT1 method. A consecutive series of 49 patients with treated myeloid malignancies and with an ELN WT1 quantitation of < 10 copies were included in the study. All cases (47 AML and 2 MDS) have received intensive chemotherapy or HCT. One to four micrograms of total RNA were retrotranscribed to obtain >= 10,000 ABL1 copies using the ELN protocol. Only those cases with a good quality cDNA were used in the ddPCR WT1 test. The ddPCR Gene Expression WT1 Assay of Bio-Rad (c) was used to perform the PCR amplification, and the microdroplets were quantified in the Bio-Rad's QX200 droplet reader. Eighteen patients showed a negative WT1 ddPCR assay (0 copies/mu l), whereas 31 cases were positive (results ranged from 1 to 15.2 copies/mu l). Survival analysis showed statistically significant differences in terms of OS between both groups, 83 +/- 8% vs. 46 +/- 9% (p = 0.024). A statistically significant correlation was also found between ddPCRWT1 results and CD123+ cell number detected by flow cytometry (p = 0.024). Larger series of patients tested with the current ddPCRWT1 method will solve whether it could be used to stratify patients with myeloid malignancies achieving deep WT1 molecular response (< 10 copies).
Filiaciones:
Bussaglia, E:
Univ Autonoma Barcelona, Hosp Santa Creu & St Pau, Hematol Dept, Hematol Lab, Barcelona 9008041, Spain
Hosp Santa Creu & Sant Pau, Hematol Dept, Barcelona, Spain
Pratcorona, M:
Univ Autonoma Barcelona, Hosp Santa Creu & St Pau, Hematol Dept, Hematol Lab, Barcelona 9008041, Spain
Hosp Santa Creu & Sant Pau, Hematol Dept, Barcelona, Spain
Carricondo, M:
Univ Autonoma Barcelona, Hosp Santa Creu & St Pau, Hematol Dept, Hematol Lab, Barcelona 9008041, Spain
Hosp Santa Creu & Sant Pau, Hematol Dept, Barcelona, Spain
Sansegundo, L:
Hosp Santa Creu & Sant Pau, Hematol Dept, Barcelona, Spain
Univ Barcelona, Barcelona, Spain
Rubio, MA:
Univ Autonoma Barcelona, Hosp Santa Creu & St Pau, Hematol Dept, Hematol Lab, Barcelona 9008041, Spain
Hosp Santa Creu & Sant Pau, Hematol Dept, Barcelona, Spain
Monter, A:
Univ Autonoma Barcelona, Hosp Santa Creu & St Pau, Hematol Dept, Hematol Lab, Barcelona 9008041, Spain
Hosp Santa Creu & Sant Pau, Hematol Dept, Barcelona, Spain
Brell, A:
Univ Autonoma Barcelona, Hosp Santa Creu & St Pau, Hematol Dept, Hematol Lab, Barcelona 9008041, Spain
Hosp Santa Creu & Sant Pau, Hematol Dept, Barcelona, Spain
Badell, I:
Hosp Santa Creu & Sant Pau, Hematol Dept, Barcelona, Spain
Hosp Santa Creu & Sant Pau, Pediat Dept, Barcelona, Spain
Esteve, J:
Hosp Clin Barcelona, Hematol Dept, Barcelona, Spain
Arnan, M:
ICO Hosp, Hematol Dept, Lhospitalet De Llobregat, Spain
Talarn, C:
Hosp Joan 23, Hematol Dept, Tarragona, Spain
Tormo, M:
Hosp Clin, Hematol Dept, Valencia, Spain
Garcia, A:
Hosp Arnau Vilanova, Hematol Dept, Lleida, Spain
Vall-Llovera, F:
Hosp Mutua Terrassa, Hematol Dept, Terrassa, Spain
Ortin, X:
Hosp Verge La Cinta, Hematol Dept, Tortosa, Spain
Pedro, C:
Parc Salut Mar, Hematol Dept, Barcelona, Spain
Bargay, J:
Hosp Son Llatzer, Hematol Dept, Palma De Mallorca, Spain
Brunet, S:
Hosp Santa Creu & Sant Pau, Hematol Dept, Barcelona, Spain
Univ Barcelona, Barcelona, Spain
Sierra, J:
Hosp Santa Creu & Sant Pau, Hematol Dept, Barcelona, Spain
Univ Barcelona, Barcelona, Spain
Nomdedeu, J:
Univ Autonoma Barcelona, Hosp Santa Creu & St Pau, Hematol Dept, Hematol Lab, Barcelona 9008041, Spain
Hosp Santa Creu & Sant Pau, Hematol Dept, Barcelona, Spain
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