title: Annotation of Sequence Variants in Cancer Samples: Processes and Pitfalls for Routine Assays in the Clinical Laboratory.
abstract: As DNA sequencing of multigene panels becomes routine for cancer samples in the clinical laboratory, an efficient process for classifying variants has become more critical. Determining which germline variants are significant for cancer disposition and which somatic mutations are integral to cancer development or therapy response remains difficult, even for well-studied genes such as BRCA1 and TP53. We compare and contrast the general principles and lines of evidence commonly used to distinguish the significance of cancer-associated germline and somatic genetic variants. The factors important in each step of the analysis pipeline are reviewed, as are some of the publicly available annotation tools. Given the range of indications and uses of cancer sequencing assays, including diagnosis, staging, prognostication, theranostics, and residual disease detection, the need for flexible methods for scoring of variants is discussed. The usefulness of protein prediction tools and multimodal risk-based or Bayesian approaches are highlighted. Using TET2 variants encountered in hematologic neoplasms, several examples of this multifactorial approach to classifying sequence variants of unknown significance are presented. Although there are still significant gaps in the publicly available data for many cancer genes that limit the broad application of explicit algorithms for variant scoring, the elements of a more rigorous model are outlined.
authors.affiliation: Department of Pathology, Quest Diagnostics Nichols Institute, Chantilly, Virginia. Electronic address: lobin.a.lee@questdiagnostics.com.
authors.forename: Lobin A
authors.initials: LA
authors.lastname: Lee
authors.affiliation: Department of Pathology, Quest Diagnostics Nichols Institute, Chantilly, Virginia.
authors.forename: Kevin J
authors.initials: KJ
authors.lastname: Arvai
authors.affiliation: Department of Pathology, Quest Diagnostics Nichols Institute, Chantilly, Virginia.
authors.forename: Dan
authors.initials: D
authors.lastname: Jones
mesh_terms: D064790:Clinical Laboratory Services; D003955:Diagnostic Tests, Routine / Q000639:trends*; D020022:Genetic Predisposition to Disease; D005820:Genetic Testing; D014644:Genetic Variation / Q000235:genetics*; D015894:Genome, Human; D006801:Humans; D058977:Molecular Sequence Annotation; D009369:Neoplasms / Q000175:diagnosis / Q000235:genetics*; D017422:Sequence Analysis, DNA / Q000379:methods*