title: Clonal architecture of CXCR4 WHIM-like mutations in Waldenström Macroglobulinaemia.
abstract: CXCR4(WHIM) somatic mutations are distinctive to Waldenström Macroglobulinaemia (WM), and impact disease presentation and treatment outcome. The clonal architecture of CXCR4(WHIM) mutations remains to be delineated. We developed highly sensitive allele-specific polymerase chain reaction (AS-PCR) assays for detecting the most common CXCR4(WHIM) mutations (CXCR4(S338X C>A and C>G) ) in WM. The AS-PCR assays detected CXCR4(S338X) mutations in WM and IgM monoclonal gammopathy of unknown significance (MGUS) patients not revealed by Sanger sequencing. By combined AS-PCR and Sanger sequencing, CXCR4(WHIM) mutations were identified in 44/102 (43%), 21/62 (34%), 2/12 (17%) and 1/20 (5%) untreated WM, previously treated WM, IgM MGUS and marginal zone lymphoma patients, respectively, but no chronic lymphocytic leukaemia, multiple myeloma, non-IgM MGUS patients or healthy donors. Cancer cell fraction analysis in WM and IgM MGUS patients showed CXCR4(S338X) mutations were primarily subclonal, with highly variable clonal distribution (median 35·1%, range 1·2-97·5%). Combined AS-PCR and Sanger sequencing revealed multiple CXCR4(WHIM) mutations in many individual WM patients, including homozygous and compound heterozygous mutations validated by deep RNA sequencing. The findings show that CXCR4(WHIM) mutations are more common in WM than previously revealed, and are primarily subclonal, supporting their acquisition after MYD88(L265P) in WM oncogenesis. The presence of multiple CXCR4(WHIM) mutations within individual WM patients may be indicative of targeted CXCR4 genomic instability.
authors.affiliation: Bing Center for Waldenström's Macroglobulinemia, Dana Farber Cancer Institute, Boston, MA, USA.
authors.forename: Lian
authors.initials: L
authors.lastname: Xu
authors.affiliation: Bing Center for Waldenström's Macroglobulinemia, Dana Farber Cancer Institute, Boston, MA, USA.|Department of Medicine, Harvard Medical School, Boston, MA, USA.
authors.forename: Zachary R
authors.initials: ZR
authors.lastname: Hunter
authors.affiliation: Bing Center for Waldenström's Macroglobulinemia, Dana Farber Cancer Institute, Boston, MA, USA.
authors.forename: Nicholas
authors.initials: N
authors.lastname: Tsakmaklis
authors.affiliation: Bing Center for Waldenström's Macroglobulinemia, Dana Farber Cancer Institute, Boston, MA, USA.|Department of Medicine, Harvard Medical School, Boston, MA, USA.
authors.forename: Yang
authors.initials: Y
authors.lastname: Cao
authors.affiliation: Bing Center for Waldenström's Macroglobulinemia, Dana Farber Cancer Institute, Boston, MA, USA.|Department of Medicine, Harvard Medical School, Boston, MA, USA.
authors.forename: Guang
authors.initials: G
authors.lastname: Yang
authors.affiliation: Bing Center for Waldenström's Macroglobulinemia, Dana Farber Cancer Institute, Boston, MA, USA.
authors.forename: Jie
authors.initials: J
authors.lastname: Chen
authors.affiliation: Bing Center for Waldenström's Macroglobulinemia, Dana Farber Cancer Institute, Boston, MA, USA.|Department of Medicine, Harvard Medical School, Boston, MA, USA.
authors.forename: Xia
authors.initials: X
authors.lastname: Liu
authors.affiliation: Bing Center for Waldenström's Macroglobulinemia, Dana Farber Cancer Institute, Boston, MA, USA.
authors.forename: Sandra
authors.initials: S
authors.lastname: Kanan
authors.affiliation: Bing Center for Waldenström's Macroglobulinemia, Dana Farber Cancer Institute, Boston, MA, USA.|Department of Medicine, Harvard Medical School, Boston, MA, USA.
authors.forename: Jorge J
authors.initials: JJ
authors.lastname: Castillo
authors.affiliation: Department of Medicine, Harvard Medical School, Boston, MA, USA.|Lipper Center for Multiple Myeloma, Dana Farber Cancer Institute, Boston, MA, USA.
authors.forename: Yu-Tzu
authors.initials: YT
authors.lastname: Tai
authors.affiliation: Department of Pathology, Stanford University Medical Center, Stanford, CA, USA.
authors.forename: James L
authors.initials: JL
authors.lastname: Zehnder
authors.affiliation: Department of Medicine, Harvard Medical School, Boston, MA, USA.|Department of Medical Oncology, Dana Farber Cancer Institute, Boston, MA, USA.
authors.forename: Jennifer R
authors.initials: JR
authors.lastname: Brown
authors.affiliation: Department of Medicine, Harvard Medical School, Boston, MA, USA.|Department of Medical Oncology, Dana Farber Cancer Institute, Boston, MA, USA.
authors.forename: Ruben D
authors.initials: RD
authors.lastname: Carrasco
authors.affiliation: Department of Medical Oncology, Stanford University Medical Center, Stanford, CA, USA.
authors.forename: Ranjana
authors.initials: R
authors.lastname: Advani
authors.affiliation: Department of Medical Oncology, Stanford University Medical Center, Stanford, CA, USA.
authors.forename: Jean M
authors.initials: JM
authors.lastname: Sabile
authors.affiliation: Department of Medical Oncology, Memorial Sloane Kettering Cancer Center, New York, NY, USA.
authors.forename: Kimon
authors.initials: K
authors.lastname: Argyropoulos
authors.affiliation: Department of Medical Oncology, Memorial Sloane Kettering Cancer Center, New York, NY, USA.
authors.forename: M
authors.initials: M
authors.lastname: Lia Palomba
authors.affiliation: Haematology Unit, Niguarda Hospital, Milan, Italy.
authors.forename: Enrica
authors.initials: E
authors.lastname: Morra
authors.affiliation: Haematology Unit, Niguarda Hospital, Milan, Italy.
authors.forename: Alessandra
authors.initials: A
authors.lastname: Trojani
authors.affiliation: Haematology Unit, Niguarda Hospital, Milan, Italy.
authors.forename: Antonino
authors.initials: A
authors.lastname: Greco
authors.affiliation: Haematology Unit, Niguarda Hospital, Milan, Italy.
authors.forename: Alessandra
authors.initials: A
authors.lastname: Tedeschi
authors.affiliation: Department of Molecular Medicine, University of Pavia, Pavia, Italy.|Department of Haematology Oncology, Fondazione IRCCS Policlinico San Matteo, Pavia, Italy.
authors.forename: Marzia
authors.initials: M
authors.lastname: Varettoni
authors.affiliation: Department of Molecular Medicine, University of Pavia, Pavia, Italy.|Department of Haematology Oncology, Fondazione IRCCS Policlinico San Matteo, Pavia, Italy.
authors.forename: Luca
authors.initials: L
authors.lastname: Arcaini
authors.affiliation: Department of Medicine, Harvard Medical School, Boston, MA, USA.|Lipper Center for Multiple Myeloma, Dana Farber Cancer Institute, Boston, MA, USA.
authors.forename: Nikhil M
authors.initials: NM
authors.lastname: Munshi
authors.affiliation: Department of Medicine, Harvard Medical School, Boston, MA, USA.|Lipper Center for Multiple Myeloma, Dana Farber Cancer Institute, Boston, MA, USA.
authors.forename: Kenneth C
authors.initials: KC
authors.lastname: Anderson
authors.affiliation: Bing Center for Waldenström's Macroglobulinemia, Dana Farber Cancer Institute, Boston, MA, USA.|Department of Medicine, Harvard Medical School, Boston, MA, USA.
authors.forename: Steven P
authors.initials: SP
authors.lastname: Treon
mesh_terms: D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D019943:Amino Acid Substitution; D005260:Female; D042822:Genomic Instability; D006801:Humans; D007075:Immunoglobulin M / Q000097:blood; D018442:Lymphoma, B-Cell, Marginal Zone / Q000235:genetics; D008297:Male; D008875:Middle Aged; D008998:Monoclonal Gammopathy of Undetermined Significance / Q000235:genetics; D009154:Mutation; D053594:Myeloid Differentiation Factor 88 / Q000235:genetics; D016133:Polymerase Chain Reaction / Q000379:methods; D019718:Receptors, CXCR4 / Q000235:genetics*; D017422:Sequence Analysis, DNA / Q000379:methods; D008258:Waldenstrom Macroglobulinemia / Q000235:genetics*
chemical_list: C527276:CXCR4 protein, human; D007075:Immunoglobulin M; C507779:MYD88 protein, human; D053594:Myeloid Differentiation Factor 88; D019718:Receptors, CXCR4
keywords: CXCR4; IgM MGUS; MYD88 L265P; Marginal Zone Lymphoma; WHIM; Waldenström macroglobulinaemia