Clinical Grade “SNaPshot” Genetic Mutation Profiling in Multiple Myeloma

Abstract

Whole genome sequencing studies have identified several oncogenic mutations in multiple myeloma (MM). As MM progresses, it evolves genetically underscoring the need to have tools for rapid detection of targetable mutations to optimize individualized treatment. Massachusetts General Hospital (MGH) has developed a Clinical Laboratory Improvement Amendments (CLIA)-approved, high-throughput, genotyping platform to determine the mutation status of a panel of known oncogenes. Sequence analysis using SNaPshot on DNA extracted from bone marrow and extramedullary plasmacytomas is feasible and leads to the detection of potentially druggable mutations. Screening MM patients for somatic mutations in oncogenes may provide novel targets leading to additional therapies for this patient population.

RET Solvent Front Mutations Mediate Acquired Resistance to Selective RET Inhibition in RET-Driven Malignancies

“…The red circles indicate the dura-based left parieto-occipital metastasis (third row), which showed significant improvement at 3 months, but progressed with increase in size at 11 months; (D) Sanger sequencing of the patient’s biopsy samples before and after selpercatinib, demonstrating a CCDC6-RET-acquired RET G810S mutation with wild-type gatekeeper residue V804. Next-generation sequencing analysis (“SNaPshot” 25 ) confirmed the RET G810S mutation and CDKN2A loss but no other acquired mutations (data not shown). IAC, Internal Auditory Canal protocol; Val, valine; Gly, glycine; Ser, serine; CT, computed tomography; RET, rearranged in transfection.…”

A DNA target-enrichment approach to detect mutations, copy number changes and immunoglobulin translocations in multiple myeloma

“… 16 Targeted NGS has significant advantages over whole-genome or whole-exome sequencing as it allows high-throughput, robust and easy analysis of chromosomal and gene lesions of large cohorts of patients by reducing the footprint of the genome to be sequenced in each case. Such studies have already been performed in acute myeloid leukemia, 17 , 18 myelodysplastic syndrome 19 , 20 and myeloma to detect recurrent gene lesions 21 , 22 or characterize immunoglobulin heavy chain (IGH) translocations, 23 but their full potential to comprehensively annotate the extended spectrum of genomic lesions with prognostic significance in myeloma has not been exploited so far.…”

SNaPshot as a Valuable Option for the Identification of Mutations in Myeloma

“…O'Donnell and colleagues reported the interest of a Clinical Laboratory Improvement Amendments-approved, high-throughput, genotyping platform to determine the mutation status of a panel of known cancer genes in MM ( O'Donnell et al, 2014 ). The method uses a highly sensitive multiplexed PCR-based assay to simultaneously identify 70 genetic loci frequently mutated in 15 cancer genes including NRAS, KRAS, TP53, BRAF and HRAS.…”

“…The assay developed by O'Donnell et al (2014) appears useful for rapid identification of mutations representing potential therapeutic targets in tumors with complex clonal evolution. Development of patient-specific personalized therapy may limit the side effects of treatment, improving compliance with dosing regimens and overall quality of life.…”

“…The SNaPshot assay reported by O'Donnell and colleagues is undoubtedly useful in clinical practice ( O'Donnell et al, 2014 ). This ready-to-use assay to detect major mutations in MM is an interesting method to integrate rapid genomic analysis into clinical routine for myeloma patients and could be valuable for adapting targeted treatment according to clonal evolution, during progression of the disease, in combination with existing therapies.…”