Digital PCR improves the quantitation of DMR and the selection of CML candidates to TKIs discontinuation

Simona Bernardi, Michele Malagola, Camilla Zanaglio, Nicola Polverelli, Elif Dereli Eke, Mariella D’Adda, Mirko Farina, Cristina Bucelli, Luigi Scaffidi, Eleonora Toffoletti, Clara Deambrogi, Fabio Stagno, Micaela Bergamaschi, Luca Franceschini, Elisabetta Abruzzese, Maria Domenica Divona, Marco Gobbi, Francesco Di Raimondo, Gianluca Gaidano, Mario Tiribelli & 4 others Massimiliano Bonifacio, Chiara Cattaneo, Alessandra Iurlo, Domenico Russo

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

Treatment-free remission (TFR) by tyrosine kinase inhibitors (TKI) discontinuation in patients with deep molecular response (DMR) is a paramount goal in the current chronic myeloid leukemia (CML) therapeutic strategy. The best DMR level by real-time quantitative PCR (RT-qPCR) for TKI discontinuation is still a matter of debate. To compare the accuracy of digital PCR (dPCR) and RT-qPCR for BCR-ABL1 transcript levels detection, 142 CML patients were monitored for a median time of 24 months. Digital PCR detected BCR-ABL1 transcripts in the RT-qPCR undetectable cases. The dPCR analysis of the samples, grouped by the MR classes, revealed a significant difference between MR4.0 and MR4.5 (P = 0.0104) or MR5.0 (P = 0.0032). The clinical and hematological characteristics of the patients grouped according to DMR classes (MR4.0 vs MR4.5-5.0) were superimposable. Conversely, patients with dPCR values <0.468 BCR-ABL1 copies/µL (as we previously described) showed a longer DMR duration (P = 0.0220) and mainly belonged to MR4.5-5.0 (P = 0.0442) classes compared to patients with higher dPCR values. Among the 142 patients, 111 (78%) discontinued the TKI treatment; among the 111 patients, 24 (22%) lost the MR3.0or MR4.0. RT-qPCR was not able to discriminate patients with higher risk of MR loss after discontinuation (P = 0.8100). On the contrary, according to dPCR, 12/25 (48%) patients with BCR-ABL1 values ≥0.468 and 12/86 (14%) patients with BCR-ABL1 values <0.468 lost DMR in this cohort, respectively (P = 0.0003). Treatment-free remission of patients who discontinued TKI with a dPCR <0.468 was significantly higher compared to patients with dPCR ≥ 0.468 (TFR at 2 years 83% vs 52% P = 0.0017, respectively). In conclusion, dPCR resulted in an improved recognition of stable DMR and of candidates to TKI discontinuation.

Original languageEnglish
Pages (from-to)2041-2055
Number of pages15
JournalCancer Medicine
Volume8
Issue number5
DOIs
Publication statusPublished - May 1 2019

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Leukemia, Myelogenous, Chronic, BCR-ABL Positive
Polymerase Chain Reaction
Protein-Tyrosine Kinases
Real-Time Polymerase Chain Reaction
Therapeutics

Keywords

  • chronic myeloid leukemia
  • digital PCR (dPCR)
  • minimal residual disease (MRD) monitoring
  • treatment-free remission (TFR)
  • tyrosine kinase inhibitors (TKI) discontinuation

ASJC Scopus subject areas

  • Oncology
  • Radiology Nuclear Medicine and imaging
  • Cancer Research

Cite this

Bernardi, S., Malagola, M., Zanaglio, C., Polverelli, N., Dereli Eke, E., D’Adda, M., ... Russo, D. (2019). Digital PCR improves the quantitation of DMR and the selection of CML candidates to TKIs discontinuation. Cancer Medicine, 8(5), 2041-2055. https://doi.org/10.1002/cam4.2087

Digital PCR improves the quantitation of DMR and the selection of CML candidates to TKIs discontinuation. / Bernardi, Simona; Malagola, Michele; Zanaglio, Camilla; Polverelli, Nicola; Dereli Eke, Elif; D’Adda, Mariella; Farina, Mirko; Bucelli, Cristina; Scaffidi, Luigi; Toffoletti, Eleonora; Deambrogi, Clara; Stagno, Fabio; Bergamaschi, Micaela; Franceschini, Luca; Abruzzese, Elisabetta; Divona, Maria Domenica; Gobbi, Marco; Di Raimondo, Francesco; Gaidano, Gianluca; Tiribelli, Mario; Bonifacio, Massimiliano; Cattaneo, Chiara; Iurlo, Alessandra; Russo, Domenico.

In: Cancer Medicine, Vol. 8, No. 5, 01.05.2019, p. 2041-2055.

Research output: Contribution to journalArticle

Bernardi, S, Malagola, M, Zanaglio, C, Polverelli, N, Dereli Eke, E, D’Adda, M, Farina, M, Bucelli, C, Scaffidi, L, Toffoletti, E, Deambrogi, C, Stagno, F, Bergamaschi, M, Franceschini, L, Abruzzese, E, Divona, MD, Gobbi, M, Di Raimondo, F, Gaidano, G, Tiribelli, M, Bonifacio, M, Cattaneo, C, Iurlo, A & Russo, D 2019, 'Digital PCR improves the quantitation of DMR and the selection of CML candidates to TKIs discontinuation', Cancer Medicine, vol. 8, no. 5, pp. 2041-2055. https://doi.org/10.1002/cam4.2087
Bernardi S, Malagola M, Zanaglio C, Polverelli N, Dereli Eke E, D’Adda M et al. Digital PCR improves the quantitation of DMR and the selection of CML candidates to TKIs discontinuation. Cancer Medicine. 2019 May 1;8(5):2041-2055. https://doi.org/10.1002/cam4.2087
Bernardi, Simona ; Malagola, Michele ; Zanaglio, Camilla ; Polverelli, Nicola ; Dereli Eke, Elif ; D’Adda, Mariella ; Farina, Mirko ; Bucelli, Cristina ; Scaffidi, Luigi ; Toffoletti, Eleonora ; Deambrogi, Clara ; Stagno, Fabio ; Bergamaschi, Micaela ; Franceschini, Luca ; Abruzzese, Elisabetta ; Divona, Maria Domenica ; Gobbi, Marco ; Di Raimondo, Francesco ; Gaidano, Gianluca ; Tiribelli, Mario ; Bonifacio, Massimiliano ; Cattaneo, Chiara ; Iurlo, Alessandra ; Russo, Domenico. / Digital PCR improves the quantitation of DMR and the selection of CML candidates to TKIs discontinuation. In: Cancer Medicine. 2019 ; Vol. 8, No. 5. pp. 2041-2055.
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abstract = "Treatment-free remission (TFR) by tyrosine kinase inhibitors (TKI) discontinuation in patients with deep molecular response (DMR) is a paramount goal in the current chronic myeloid leukemia (CML) therapeutic strategy. The best DMR level by real-time quantitative PCR (RT-qPCR) for TKI discontinuation is still a matter of debate. To compare the accuracy of digital PCR (dPCR) and RT-qPCR for BCR-ABL1 transcript levels detection, 142 CML patients were monitored for a median time of 24 months. Digital PCR detected BCR-ABL1 transcripts in the RT-qPCR undetectable cases. The dPCR analysis of the samples, grouped by the MR classes, revealed a significant difference between MR4.0 and MR4.5 (P = 0.0104) or MR5.0 (P = 0.0032). The clinical and hematological characteristics of the patients grouped according to DMR classes (MR4.0 vs MR4.5-5.0) were superimposable. Conversely, patients with dPCR values <0.468 BCR-ABL1 copies/µL (as we previously described) showed a longer DMR duration (P = 0.0220) and mainly belonged to MR4.5-5.0 (P = 0.0442) classes compared to patients with higher dPCR values. Among the 142 patients, 111 (78{\%}) discontinued the TKI treatment; among the 111 patients, 24 (22{\%}) lost the MR3.0or MR4.0. RT-qPCR was not able to discriminate patients with higher risk of MR loss after discontinuation (P = 0.8100). On the contrary, according to dPCR, 12/25 (48{\%}) patients with BCR-ABL1 values ≥0.468 and 12/86 (14{\%}) patients with BCR-ABL1 values <0.468 lost DMR in this cohort, respectively (P = 0.0003). Treatment-free remission of patients who discontinued TKI with a dPCR <0.468 was significantly higher compared to patients with dPCR ≥ 0.468 (TFR at 2 years 83{\%} vs 52{\%} P = 0.0017, respectively). In conclusion, dPCR resulted in an improved recognition of stable DMR and of candidates to TKI discontinuation.",
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T1 - Digital PCR improves the quantitation of DMR and the selection of CML candidates to TKIs discontinuation

AU - Bernardi, Simona

AU - Malagola, Michele

AU - Zanaglio, Camilla

AU - Polverelli, Nicola

AU - Dereli Eke, Elif

AU - D’Adda, Mariella

AU - Farina, Mirko

AU - Bucelli, Cristina

AU - Scaffidi, Luigi

AU - Toffoletti, Eleonora

AU - Deambrogi, Clara

AU - Stagno, Fabio

AU - Bergamaschi, Micaela

AU - Franceschini, Luca

AU - Abruzzese, Elisabetta

AU - Divona, Maria Domenica

AU - Gobbi, Marco

AU - Di Raimondo, Francesco

AU - Gaidano, Gianluca

AU - Tiribelli, Mario

AU - Bonifacio, Massimiliano

AU - Cattaneo, Chiara

AU - Iurlo, Alessandra

AU - Russo, Domenico

PY - 2019/5/1

Y1 - 2019/5/1

N2 - Treatment-free remission (TFR) by tyrosine kinase inhibitors (TKI) discontinuation in patients with deep molecular response (DMR) is a paramount goal in the current chronic myeloid leukemia (CML) therapeutic strategy. The best DMR level by real-time quantitative PCR (RT-qPCR) for TKI discontinuation is still a matter of debate. To compare the accuracy of digital PCR (dPCR) and RT-qPCR for BCR-ABL1 transcript levels detection, 142 CML patients were monitored for a median time of 24 months. Digital PCR detected BCR-ABL1 transcripts in the RT-qPCR undetectable cases. The dPCR analysis of the samples, grouped by the MR classes, revealed a significant difference between MR4.0 and MR4.5 (P = 0.0104) or MR5.0 (P = 0.0032). The clinical and hematological characteristics of the patients grouped according to DMR classes (MR4.0 vs MR4.5-5.0) were superimposable. Conversely, patients with dPCR values <0.468 BCR-ABL1 copies/µL (as we previously described) showed a longer DMR duration (P = 0.0220) and mainly belonged to MR4.5-5.0 (P = 0.0442) classes compared to patients with higher dPCR values. Among the 142 patients, 111 (78%) discontinued the TKI treatment; among the 111 patients, 24 (22%) lost the MR3.0or MR4.0. RT-qPCR was not able to discriminate patients with higher risk of MR loss after discontinuation (P = 0.8100). On the contrary, according to dPCR, 12/25 (48%) patients with BCR-ABL1 values ≥0.468 and 12/86 (14%) patients with BCR-ABL1 values <0.468 lost DMR in this cohort, respectively (P = 0.0003). Treatment-free remission of patients who discontinued TKI with a dPCR <0.468 was significantly higher compared to patients with dPCR ≥ 0.468 (TFR at 2 years 83% vs 52% P = 0.0017, respectively). In conclusion, dPCR resulted in an improved recognition of stable DMR and of candidates to TKI discontinuation.

AB - Treatment-free remission (TFR) by tyrosine kinase inhibitors (TKI) discontinuation in patients with deep molecular response (DMR) is a paramount goal in the current chronic myeloid leukemia (CML) therapeutic strategy. The best DMR level by real-time quantitative PCR (RT-qPCR) for TKI discontinuation is still a matter of debate. To compare the accuracy of digital PCR (dPCR) and RT-qPCR for BCR-ABL1 transcript levels detection, 142 CML patients were monitored for a median time of 24 months. Digital PCR detected BCR-ABL1 transcripts in the RT-qPCR undetectable cases. The dPCR analysis of the samples, grouped by the MR classes, revealed a significant difference between MR4.0 and MR4.5 (P = 0.0104) or MR5.0 (P = 0.0032). The clinical and hematological characteristics of the patients grouped according to DMR classes (MR4.0 vs MR4.5-5.0) were superimposable. Conversely, patients with dPCR values <0.468 BCR-ABL1 copies/µL (as we previously described) showed a longer DMR duration (P = 0.0220) and mainly belonged to MR4.5-5.0 (P = 0.0442) classes compared to patients with higher dPCR values. Among the 142 patients, 111 (78%) discontinued the TKI treatment; among the 111 patients, 24 (22%) lost the MR3.0or MR4.0. RT-qPCR was not able to discriminate patients with higher risk of MR loss after discontinuation (P = 0.8100). On the contrary, according to dPCR, 12/25 (48%) patients with BCR-ABL1 values ≥0.468 and 12/86 (14%) patients with BCR-ABL1 values <0.468 lost DMR in this cohort, respectively (P = 0.0003). Treatment-free remission of patients who discontinued TKI with a dPCR <0.468 was significantly higher compared to patients with dPCR ≥ 0.468 (TFR at 2 years 83% vs 52% P = 0.0017, respectively). In conclusion, dPCR resulted in an improved recognition of stable DMR and of candidates to TKI discontinuation.

KW - chronic myeloid leukemia

KW - digital PCR (dPCR)

KW - minimal residual disease (MRD) monitoring

KW - treatment-free remission (TFR)

KW - tyrosine kinase inhibitors (TKI) discontinuation

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