Acquired mutations associated with ibrutinib resistance in Waldenström macroglobulinemia.

Overview

abstract

Ibrutinib produces high response rates and durable remissions in Waldenström macroglobulinemia (WM) that are impacted by MYD88 and CXCR4^WHIM mutations. Disease progression can develop on ibrutinib, although the molecular basis remains to be clarified. We sequenced sorted CD19⁺ lymphoplasmacytic cells from 6 WM patients who progressed after achieving major responses on ibrutinib using Sanger, TA cloning and sequencing, and highly sensitive and allele-specific polymerase chain reaction (AS-PCR) assays that we developed for Bruton tyrosine kinase (BTK) mutations. AS-PCR assays were used to screen patients with and without progressive disease on ibrutinib, and ibrutinib-naïve disease. Targeted next-generation sequencing was used to validate AS-PCR findings, assess for other BTK mutations, and other targets in B-cell receptor and MYD88 signaling. Among the 6 progressing patients, 3 had BTK^Cys481 variants that included BTK^{Cys481Ser(c.1635G>C and c.1634T>A)} and BTK^{Cys481Arg(c.1634T>C)} Two of these patients had multiple BTK mutations. Screening of 38 additional patients on ibrutinib without clinical progression identified BTK^Cys481 mutations in 2 (5.1%) individuals, both of whom subsequently progressed. BTK^Cys481 mutations were not detected in baseline samples or in 100 ibrutinib-naive WM patients. Using mutated MYD88 as a tumor marker, BTK^Cys481 mutations were subclonal, with a highly variable clonal distribution. Targeted deep-sequencing confirmed AS-PCR findings, and identified an additional BTK^{Cys481Tyr(c.1634G>A)} mutation in the 2 patients with multiple other BTK^Cys481 mutations, as well as CARD11^{Leu878Phe(c.2632C>T)} and PLCγ2^{Tyr495His(c.1483T>C)} mutations. Four of the 5 patients with BTK^C481 variants were CXCR4 mutated. BTK^Cys481 mutations are common in WM patients with clinical progression on ibrutinib, and are associated with mutated CXCR4.