title: Targeted cancer exome sequencing reveals recurrent mutations in myeloproliferative neoplasms.
abstract: With the intent of dissecting the molecular complexity of Philadelphia-negative myeloproliferative neoplasms (MPN), we designed a target enrichment panel to explore, using next-generation sequencing (NGS), the mutational status of an extensive list of 2000 cancer-associated genes and microRNAs. The genomic DNA of granulocytes and in vitro-expanded CD3+T-lymphocytes, as a germline control, was target-enriched and sequenced in a learning cohort of 20 MPN patients using Roche 454 technology. We identified 141 genuine somatic mutations, most of which were not previously described. To test the frequency of the identified variants, a larger validation cohort of 189 MPN patients was additionally screened for these mutations using Ion Torrent AmpliSeq NGS. Excluding the genes already described in MPN, for 8 genes (SCRIB, MIR662, BARD1, TCF12, FAT4, DAP3, POLG and NRAS), we demonstrated a mutation frequency between 3 and 8%. We also found that mutations at codon 12 of NRAS (NRASG12V and NRASG12D) were significantly associated, for primary myelofibrosis (PMF), with highest dynamic international prognostic scoring system (DIPSS)-plus score categories. This association was then confirmed in 66 additional PMF patients composing a final dataset of 168 PMF showing a NRAS mutation frequency of 4.7%, which was associated with a worse outcome, as defined by the DIPSS plus score.
authors.affiliation: 1] Center for Genome Research, University of Modena and Reggio Emilia, Modena, Italy [2] Life Sciences Department, University of Modena and Reggio Emilia, Modena, Italy [3] Center for Regenerative Medicine 'Stefano Ferrari', University of Modena and Reggio Emilia, Modena, Italy.
authors.forename: E
authors.initials: E
authors.lastname: Tenedini
authors.affiliation: 1] Center for Genome Research, University of Modena and Reggio Emilia, Modena, Italy [2] Life Sciences Department, University of Modena and Reggio Emilia, Modena, Italy.
authors.forename: I
authors.initials: I
authors.lastname: Bernardis
authors.affiliation: 1] Center for Genome Research, University of Modena and Reggio Emilia, Modena, Italy [2] Life Sciences Department, University of Modena and Reggio Emilia, Modena, Italy.
authors.forename: V
authors.initials: V
authors.lastname: Artusi
authors.affiliation: 1] Center for Genome Research, University of Modena and Reggio Emilia, Modena, Italy [2] Life Sciences Department, University of Modena and Reggio Emilia, Modena, Italy.
authors.forename: L
authors.initials: L
authors.lastname: Artuso
authors.affiliation: 1] Center for Genome Research, University of Modena and Reggio Emilia, Modena, Italy [2] Life Sciences Department, University of Modena and Reggio Emilia, Modena, Italy.
authors.forename: E
authors.initials: E
authors.lastname: Roncaglia
authors.affiliation: Department of Experimental and Clinical Medicine, University of Florence, Florence, Italy.
authors.forename: P
authors.initials: P
authors.lastname: Guglielmelli
authors.affiliation: Department of Experimental and Clinical Medicine, University of Florence, Florence, Italy.
authors.forename: L
authors.initials: L
authors.lastname: Pieri
authors.affiliation: Department of Experimental and Clinical Medicine, University of Florence, Florence, Italy.
authors.forename: C
authors.initials: C
authors.lastname: Bogani
authors.affiliation: Department of Experimental and Clinical Medicine, University of Florence, Florence, Italy.
authors.forename: F
authors.initials: F
authors.lastname: Biamonte
authors.affiliation: Department of Experimental and Clinical Medicine, University of Florence, Florence, Italy.
authors.forename: G
authors.initials: G
authors.lastname: Rotunno
authors.affiliation: Department of Experimental and Clinical Medicine, University of Florence, Florence, Italy.
authors.forename: C
authors.initials: C
authors.lastname: Mannarelli
authors.affiliation: 1] Life Sciences Department, University of Modena and Reggio Emilia, Modena, Italy [2] Center for Regenerative Medicine 'Stefano Ferrari', University of Modena and Reggio Emilia, Modena, Italy.
authors.forename: E
authors.initials: E
authors.lastname: Bianchi
authors.affiliation: Department of Experimental and Clinical Medicine, University of Florence, Florence, Italy.
authors.forename: A
authors.initials: A
authors.lastname: Pancrazzi
authors.affiliation: Department of Experimental and Clinical Medicine, University of Florence, Florence, Italy.
authors.forename: T
authors.initials: T
authors.lastname: Fanelli
authors.affiliation: 1] Center for Genome Research, University of Modena and Reggio Emilia, Modena, Italy [2] Life Sciences Department, University of Modena and Reggio Emilia, Modena, Italy.
authors.forename: G
authors.initials: G
authors.lastname: Malagoli Tagliazucchi
authors.affiliation: 1] Center for Genome Research, University of Modena and Reggio Emilia, Modena, Italy [2] Life Sciences Department, University of Modena and Reggio Emilia, Modena, Italy.
authors.forename: S
authors.initials: S
authors.lastname: Ferrari
authors.affiliation: 1] Life Sciences Department, University of Modena and Reggio Emilia, Modena, Italy [2] Center for Regenerative Medicine 'Stefano Ferrari', University of Modena and Reggio Emilia, Modena, Italy.
authors.forename: R
authors.initials: R
authors.lastname: Manfredini
authors.affiliation: Department of Experimental and Clinical Medicine, University of Florence, Florence, Italy.
authors.forename: A M
authors.initials: AM
authors.lastname: Vannucchi
authors.affiliation: 1] Center for Genome Research, University of Modena and Reggio Emilia, Modena, Italy [2] Life Sciences Department, University of Modena and Reggio Emilia, Modena, Italy.
authors.forename: E
authors.initials: E
authors.lastname: Tagliafico
mesh_terms: D015331:Cohort Studies; D059472:Exome; D018095:Germ-Line Mutation; D006801:Humans; D009196:Myeloproliferative Disorders / Q000235:genetics*; D009369:Neoplasms / Q000235:genetics*