TY - JOUR
T1 - Validation of the EuroClonality-NGS DNA capture panel as an integrated genomic tool for lymphoproliferative disorders
AU - Stewart, James Peter
AU - Gazdova, Jana
AU - Darzentas, Nikos
AU - Wren, Dorte
AU - Proszek, Paula
AU - Fazio, Grazia
AU - Songia, Simona
AU - Alcoceba, Miguel
AU - Sarasquete, Maria Eugenia
AU - Villarese, Patrick
AU - Van Der Klift, Michele Y.
AU - Heezen, Kim C.
AU - McCafferty, Neil
AU - Pal, Karol
AU - Catherwood, Mark
AU - Kim, Chang Sik
AU - Srivastava, Shambhavi
AU - Kroeze, Leonie I.
AU - Hodges, Elizabeth
AU - Stamatopoulos, Kostas
AU - Klapper, Wolfram
AU - Genuardi, Elisa
AU - Ferrero, Simone
AU - Van Den Brand, Michiel
AU - Cazzaniga, Giovanni
AU - Davi, Frederic
AU - Sutton, Lesley Ann
AU - Garcia-Sanz, Ramon
AU - Groenen, Patricia J.T.A.
AU - Macintyre, Elizabeth A.
AU - Bruggemann, Monika
AU - Pott, Christiane
AU - Langerak, Anton W.
AU - Gonzalez, David
N1 - Funding Information:
The Northern Ireland High Performance Computing Service provided computing resources, funded by the Engineering and Physical Sciences Research Council (EP/T022175).
Funding Information:
Conflict-of-interest disclosure: M.A. has received honoraria from Janssen and nonfinancial support (event attendance) from Janssen, AbbVie, and Roche. W.K. has received research grants from Roche, Regeneron, Amgen, and Takeda. L.-A.S. has received honoraria from Janssen. R.G.-S. has received honoraria, consultancy, and/or research funding from Amgen, Bristol-Myers Squibb, Gilead, Incyte, Janssen, Novartis, Pharmacyclics, and Takeda. E.M. provides consultancy services to Servier for minimal residual disease measurement in ALL. A.W.L. has received contract research support from Roche-Genentech, Gilead, and Janssen. D.G. has received honoraria, consultancy, and/or research funding from Roche, AstraZeneca, Novar-tis, Incyte, Eli Lilly, and Illumina and is cofounder of Univ8 Genomics. The remaining authors declare no competing financial interests.
Publisher Copyright:
© 2021 American Society of Hematology. All rights reserved.
PY - 2021/8/24
Y1 - 2021/8/24
N2 - Current diagnostic standards for lymphoproliferative disorders include multiple tests for detection of clonal immunoglobulin (IG) and/or T-cell receptor (TCR) rearrangements, translocations, copy-number alterations (CNAs), and somatic mutations. The EuroClonality-NGS DNA Capture (EuroClonality-NDC) assay was designed as an integrated tool to characterize these alterations by capturing IGH switch regions along with variable, diversity, and joining genes of all IG and TCR loci in addition to clinically relevant genes for CNA and mutation analysis. Diagnostic performance against standard-of-care clinical testing was assessed in a cohort of 280 B-and T-cell malignancies from 10 European laboratories, including 88 formalin-fixed paraffin-embedded samples and 21 reactive lesions. DNA samples were subjected to the EuroClonality-NDC protocol in 7 EuroClonality-NGS laboratories and analyzed using a bespoke bioinformatic pipeline. The EuroClonality-NDC assay detected B-cell clonality in 191 (97%) of 197 B-cell malignancies and T-cell clonality in 71 (97%) of 73 T-cell malignancies. Limit of detection (LOD) for IG/TCR rearrangements was established at 5% using cell line blends. Chromosomal translocations were detected in 145 (95%) of 152 cases known to be positive. CNAs were validated for immunogenetic and oncogenetic regions, highlighting their novel role in confirming clonality in somatically hypermutated cases. Single-nucleotide variant LOD was determined as 4% allele frequency, and an orthogonal validation using 32 samples resulted in 98% concordance. The EuroClonality-NDC assay is a robust tool providing a single end-To-end workflow for simultaneous detection of B-and T-cell clonality, translocations, CNAs, and sequence variants.
AB - Current diagnostic standards for lymphoproliferative disorders include multiple tests for detection of clonal immunoglobulin (IG) and/or T-cell receptor (TCR) rearrangements, translocations, copy-number alterations (CNAs), and somatic mutations. The EuroClonality-NGS DNA Capture (EuroClonality-NDC) assay was designed as an integrated tool to characterize these alterations by capturing IGH switch regions along with variable, diversity, and joining genes of all IG and TCR loci in addition to clinically relevant genes for CNA and mutation analysis. Diagnostic performance against standard-of-care clinical testing was assessed in a cohort of 280 B-and T-cell malignancies from 10 European laboratories, including 88 formalin-fixed paraffin-embedded samples and 21 reactive lesions. DNA samples were subjected to the EuroClonality-NDC protocol in 7 EuroClonality-NGS laboratories and analyzed using a bespoke bioinformatic pipeline. The EuroClonality-NDC assay detected B-cell clonality in 191 (97%) of 197 B-cell malignancies and T-cell clonality in 71 (97%) of 73 T-cell malignancies. Limit of detection (LOD) for IG/TCR rearrangements was established at 5% using cell line blends. Chromosomal translocations were detected in 145 (95%) of 152 cases known to be positive. CNAs were validated for immunogenetic and oncogenetic regions, highlighting their novel role in confirming clonality in somatically hypermutated cases. Single-nucleotide variant LOD was determined as 4% allele frequency, and an orthogonal validation using 32 samples resulted in 98% concordance. The EuroClonality-NDC assay is a robust tool providing a single end-To-end workflow for simultaneous detection of B-and T-cell clonality, translocations, CNAs, and sequence variants.
UR - http://www.scopus.com/inward/record.url?scp=85113837183&partnerID=8YFLogxK
U2 - 10.1182/bloodadvances.2020004056
DO - 10.1182/bloodadvances.2020004056
M3 - Article
AN - SCOPUS:85113837183
VL - 5
SP - 3188
EP - 3198
JO - Blood advances
JF - Blood advances
SN - 2473-9529
IS - 16
ER -