A complementary role of multiparameter flow cytometry and high-throughput sequencing for minimal residual disease detection in chronic lymphocytic leukemia: an European Research Initiative on CLL study

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Authors	RAWSTRON A.C. FAZI C. AGATHANGELIDIS A. VILLAMOR N. LETESTU R. NOMDEDEU J. PALACIO C. STEHLÍKOVÁ Olga KREUZER K-A. LIPTROT S. O´BRIEN D. DE TUTE R.M. MARINOV I. HAUWEL M. SPACEK M. DOBBER J. KATER A.P. GAMBELL P. SOOSAPILLA A. LOZANSKI G. BRACHTL G. LIN K. BOYSEN J. HANSON C. JORGENSEN J.L. STETLER-STEVENSON M. YUAN C. BROOME H.E. RASSENTI L. CRAIG F. DELGADO J. MORENO C. BOSCH F. EGLE A. DOUBEK Michael POSPÍŠILOVÁ Šárka MULLIGAN S. WESTERMAN D. SANDERS C.M. EMERSON R. ROBINS H.S. KIRSCH I. SHANAFELT T. PETTITT A. KIPPS T.J. WIERDA W.G. CYMBALISTA F. HALLEK M. HILLMEN P. MONTSERRAT E. GHIA P.
Year of publication	2016
Type	Article in Periodical
Magazine / Source	Leukemia
MU Faculty or unit	Central European Institute of Technology
Citation
web	https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4832072/pdf/leu2015313a.pdf
Doi	http://dx.doi.org/10.1038/leu.2015.313
Field	Oncology and hematology
Keywords	ASSAY; QUANTIFICATION; RITUXIMAB; SURVIVAL; THERAPY; ERADICATION; EXPRESSION; GUIDELINES; DIAGNOSIS; TRIAL
Description	In chronic lymphocytic leukemia (CLL) the level of minimal residual disease (MRD) after therapy is an independent predictor of outcome. Given the increasing number of new agents being explored for CLL therapy, using MRD as a surrogate could greatly reduce the time necessary to assess their efficacy. In this European Research Initiative on CLL (ERIC) project we have identified and validated a flow-cytometric approach to reliably quantitate CLL cells to the level of 0.0010% (10(-5)). The assay comprises a core panel of six markers (i.e. CD19, CD20, CD5, CD43, CD79b and CD81) with a component specification independent of instrument and reagents, which can be locally re-validated using normal peripheral blood. This method is directly comparable to previous ERIC-designed assays and also provides a backbone for investigation of new markers. A parallel analysis of high-throughput sequencing using the ClonoSEQ assay showed good concordance with flow cytometry results at the 0.010% (10(-4)) level, the MRD threshold defined in the 2008 International Workshop on CLL guidelines, but it also provides good linearity to a detection limit of 1 in a million (10(-6)). The combination of both technologies would permit a highly sensitive approach to MRD detection while providing a reproducible and broadly accessible method to quantify residual disease and optimize treatment in CLL.
Related projects:	CEITEC - Central European Institute of Technology Molekulární charakterizace B buněčných receptorů a jejich vztah k evoluci genetických změn u chronické lymfocytární leukémie