Droplet Digital PCR Improves IG-/TR-based MRD Risk Definition in Childhood B-cell Precursor Acute Lymphoblastic Leukemia
Minimal residual disease (MRD) is the most powerful prognostic factor in pediatric acute lymphoblastic leukemia (ALL). Real-time quantitative polymerase chain reaction (RQ-PCR) represents the gold standard for molecular MRD assessment and risk-based stratification of front-line treatment. In the pro...
| Main Authors: | , , , , , , , , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
Wiley
2021-03-01
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| Series: | HemaSphere |
| Online Access: | http://journals.lww.com/10.1097/HS9.0000000000000543 |
| _version_ | 1797283322500481024 |
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| author | Irene Della Starza Vittorio Nunes Federica Lovisa Daniela Silvestri Marzia Cavalli Andrea Garofalo Mimma Campeggio Lucia Anna De Novi Roberta Soscia Carlotta Oggioni Lara Mussolin Andrea Biondi Anna Guarini Maria Grazia Valsecchi Valentino Conter Alessandra Biffi Giuseppe Basso Robin Foà Giovanni Cazzaniga |
| author_facet | Irene Della Starza Vittorio Nunes Federica Lovisa Daniela Silvestri Marzia Cavalli Andrea Garofalo Mimma Campeggio Lucia Anna De Novi Roberta Soscia Carlotta Oggioni Lara Mussolin Andrea Biondi Anna Guarini Maria Grazia Valsecchi Valentino Conter Alessandra Biffi Giuseppe Basso Robin Foà Giovanni Cazzaniga |
| author_sort | Irene Della Starza |
| collection | DOAJ |
| description | Minimal residual disease (MRD) is the most powerful prognostic factor in pediatric acute lymphoblastic leukemia (ALL). Real-time quantitative polymerase chain reaction (RQ-PCR) represents the gold standard for molecular MRD assessment and risk-based stratification of front-line treatment. In the protocols of the Italian Association of Pediatric Hematology and Oncology (AIEOP) and the Berlin-Frankfurth-Munschen (BFM) group AIEOP-BFM ALL2009 and ALL2017, B-lineage ALL patients with high RQ-PCR-MRD at day+33 and positive at day+78 are defined slow early responders (SERs). Based on results of the AIEOP-BFM ALL2000 study, these patients are treated as high-risk also when positive MRD signal at day +78 is below the lower limit of quantification of RQ-PCR (“positive not-quantifiable,” POS-NQ). To assess whether droplet digital polymerase chain reaction (ddPCR) could improve patients’ risk definition, we analyzed MRD in 209 pediatric B-lineage ALL cases classified by RQ-PCR as POS-NQ and/or negative (NEG) at days +33 and/or +78 in the AIEOP-BFM ALL2000 trial. ddPCR MRD analysis was performed on 45 samples collected at day +78 from SER patients, who had RQ-PCR MRD ≥ 5.0 × 10–4 at day+33 and POS-NQ at day+78 and were treated as medium risk (MR). The analysis identified 13 of 45 positive quantifiable cases. Most relapses occurred in this patients’ subgroup, while ddPCR NEG or ddPCR-POS-NQ patients had a significantly better outcome (P < 0.001). Overall, in 112 MR cases and 52 standard-risk patients, MRD negativity and POS-NQ were confirmed by the ddPCR analysis except for a minority of cases, for whom no differences in outcome were registered. These data indicate that ddPCR is more accurate than RQ-PCR in the measurement of MRD, particularly in late follow-up time points, and may thus allow improving patients’ stratification in ALL protocols. |
| first_indexed | 2024-03-07T17:29:25Z |
| format | Article |
| id | doaj.art-b1b6d3f45e824146b248f9d4d86c5d5c |
| institution | Directory Open Access Journal |
| issn | 2572-9241 |
| language | English |
| last_indexed | 2024-03-07T17:29:25Z |
| publishDate | 2021-03-01 |
| publisher | Wiley |
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| series | HemaSphere |
| spelling | doaj.art-b1b6d3f45e824146b248f9d4d86c5d5c2024-03-02T18:22:19ZengWileyHemaSphere2572-92412021-03-0153e54310.1097/HS9.0000000000000543202103000-00017Droplet Digital PCR Improves IG-/TR-based MRD Risk Definition in Childhood B-cell Precursor Acute Lymphoblastic LeukemiaIrene Della Starza0Vittorio Nunes1Federica Lovisa2Daniela Silvestri3Marzia Cavalli4Andrea Garofalo5Mimma Campeggio6Lucia Anna De Novi7Roberta Soscia8Carlotta Oggioni9Lara Mussolin10Andrea Biondi11Anna Guarini12Maria Grazia Valsecchi13Valentino Conter14Alessandra Biffi15Giuseppe Basso16Robin Foà17Giovanni Cazzaniga181 Hematology, Department of Translational and Precision Medicine, “Sapienza” University of Rome, Italy3 Tettamanti Research Centre, Pediatrics, University of Milano-Bicocca, Monza, Italy4 Division of Pediatric Hematology and Oncology, Department of Women’s and Children’s Health, University of Padova, Padova, Italy & Istituto di Ricerca Pediatrica Città della Speranza, Padova, Italy5 Center of Bioinformatics, Biostatistics and Bioimaging, School of Medicine and Surgery, University of Milano-Bicocca, Monza, Italy1 Hematology, Department of Translational and Precision Medicine, “Sapienza” University of Rome, Italy3 Tettamanti Research Centre, Pediatrics, University of Milano-Bicocca, Monza, Italy4 Division of Pediatric Hematology and Oncology, Department of Women’s and Children’s Health, University of Padova, Padova, Italy & Istituto di Ricerca Pediatrica Città della Speranza, Padova, Italy1 Hematology, Department of Translational and Precision Medicine, “Sapienza” University of Rome, Italy1 Hematology, Department of Translational and Precision Medicine, “Sapienza” University of Rome, Italy3 Tettamanti Research Centre, Pediatrics, University of Milano-Bicocca, Monza, Italy4 Division of Pediatric Hematology and Oncology, Department of Women’s and Children’s Health, University of Padova, Padova, Italy & Istituto di Ricerca Pediatrica Città della Speranza, Padova, Italy6 Pediatrics, Fondazione MBBM/San Gerardo Hospital, University of Milano-Bicocca, Monza, Italy1 Hematology, Department of Translational and Precision Medicine, “Sapienza” University of Rome, Italy5 Center of Bioinformatics, Biostatistics and Bioimaging, School of Medicine and Surgery, University of Milano-Bicocca, Monza, Italy6 Pediatrics, Fondazione MBBM/San Gerardo Hospital, University of Milano-Bicocca, Monza, Italy4 Division of Pediatric Hematology and Oncology, Department of Women’s and Children’s Health, University of Padova, Padova, Italy & Istituto di Ricerca Pediatrica Città della Speranza, Padova, Italy9 Italian Institute for Genomic Medicine, Torino, Italy1 Hematology, Department of Translational and Precision Medicine, “Sapienza” University of Rome, Italy3 Tettamanti Research Centre, Pediatrics, University of Milano-Bicocca, Monza, ItalyMinimal residual disease (MRD) is the most powerful prognostic factor in pediatric acute lymphoblastic leukemia (ALL). Real-time quantitative polymerase chain reaction (RQ-PCR) represents the gold standard for molecular MRD assessment and risk-based stratification of front-line treatment. In the protocols of the Italian Association of Pediatric Hematology and Oncology (AIEOP) and the Berlin-Frankfurth-Munschen (BFM) group AIEOP-BFM ALL2009 and ALL2017, B-lineage ALL patients with high RQ-PCR-MRD at day+33 and positive at day+78 are defined slow early responders (SERs). Based on results of the AIEOP-BFM ALL2000 study, these patients are treated as high-risk also when positive MRD signal at day +78 is below the lower limit of quantification of RQ-PCR (“positive not-quantifiable,” POS-NQ). To assess whether droplet digital polymerase chain reaction (ddPCR) could improve patients’ risk definition, we analyzed MRD in 209 pediatric B-lineage ALL cases classified by RQ-PCR as POS-NQ and/or negative (NEG) at days +33 and/or +78 in the AIEOP-BFM ALL2000 trial. ddPCR MRD analysis was performed on 45 samples collected at day +78 from SER patients, who had RQ-PCR MRD ≥ 5.0 × 10–4 at day+33 and POS-NQ at day+78 and were treated as medium risk (MR). The analysis identified 13 of 45 positive quantifiable cases. Most relapses occurred in this patients’ subgroup, while ddPCR NEG or ddPCR-POS-NQ patients had a significantly better outcome (P < 0.001). Overall, in 112 MR cases and 52 standard-risk patients, MRD negativity and POS-NQ were confirmed by the ddPCR analysis except for a minority of cases, for whom no differences in outcome were registered. These data indicate that ddPCR is more accurate than RQ-PCR in the measurement of MRD, particularly in late follow-up time points, and may thus allow improving patients’ stratification in ALL protocols.http://journals.lww.com/10.1097/HS9.0000000000000543 |
| spellingShingle | Irene Della Starza Vittorio Nunes Federica Lovisa Daniela Silvestri Marzia Cavalli Andrea Garofalo Mimma Campeggio Lucia Anna De Novi Roberta Soscia Carlotta Oggioni Lara Mussolin Andrea Biondi Anna Guarini Maria Grazia Valsecchi Valentino Conter Alessandra Biffi Giuseppe Basso Robin Foà Giovanni Cazzaniga Droplet Digital PCR Improves IG-/TR-based MRD Risk Definition in Childhood B-cell Precursor Acute Lymphoblastic Leukemia HemaSphere |
| title | Droplet Digital PCR Improves IG-/TR-based MRD Risk Definition in Childhood B-cell Precursor Acute Lymphoblastic Leukemia |
| title_full | Droplet Digital PCR Improves IG-/TR-based MRD Risk Definition in Childhood B-cell Precursor Acute Lymphoblastic Leukemia |
| title_fullStr | Droplet Digital PCR Improves IG-/TR-based MRD Risk Definition in Childhood B-cell Precursor Acute Lymphoblastic Leukemia |
| title_full_unstemmed | Droplet Digital PCR Improves IG-/TR-based MRD Risk Definition in Childhood B-cell Precursor Acute Lymphoblastic Leukemia |
| title_short | Droplet Digital PCR Improves IG-/TR-based MRD Risk Definition in Childhood B-cell Precursor Acute Lymphoblastic Leukemia |
| title_sort | droplet digital pcr improves ig tr based mrd risk definition in childhood b cell precursor acute lymphoblastic leukemia |
| url | http://journals.lww.com/10.1097/HS9.0000000000000543 |
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