title: Genomic Alterations of Anaplastic Thyroid Carcinoma Detected by Targeted Massive Parallel Sequencing in a BRAF(V600E) Mutation-Prevalent Area.
abstract: Anaplastic thyroid carcinoma (ATC), the most aggressive type of thyroid cancer, has no effective therapy. Due to its dismal prognosis, it is vital to understand the genetic alterations of ATC and identify effective molecular targets. Targeted next-generation sequencing was performed to investigate the mutational profile of ATC using a massive parallel sequencing approach.
  
  
  
  DNA from formalin-fixed, paraffin-embedded archival samples of 11 ATCs and normal matched pairs were used. A total of 48 genetic alterations were identified by targeted exome sequencing. These alterations were validated by mass spectrometric genotyping and direct Sanger sequencing.
  
  
  
  The most commonly mutated gene was BRAF, identified in 10 samples (91%), all showing the V600E point mutation. A KRAS point mutation was observed in the one sample (9%) without the BRAF(V600E) mutation. All 11 ATCs harbored BRAF or RAS mutations, reflecting the possibility that differentiated thyroid carcinomas progress to ATCs after the accumulation of mutations. A loss of function mutation of TP53 was observed in eight samples (73%), a PIK3CA mutation was observed in two samples (18%), and a frameshift mutation of PTEN was observed in one sample (9%). Twenty-eight novel mutated genes were found that had not previously been associated with ATC. Of these, loss of function mutations of NF2, KMT2D, and PKHD1 were repeatedly seen in three samples (27%), two samples (18%), and two samples (18%), respectively. Using direct Sanger sequencing, two samples (18%) were also found with a RASAL1 mutation. KMT2D and RASAL1 mutations were significantly associated with shorter ATC patient survival.
  
  
  
  This comprehensive analysis of ATCs using targeted massive parallel sequencing identified several novel mutations in ATCs, such as loss of function mutations of NF2 or KMT2D. Future studies are needed to confirm the role of these novel mutations as independent drivers of ATC development.
authors.affiliation: 1 Department of Internal Medicine, Asan Medical Center, University of Ulsan College of Medicine , Seoul, Korea.
authors.forename: Min Ji
authors.initials: MJ
authors.lastname: Jeon
authors.affiliation: 2 Department of Pathology, Asan Medical Center, University of Ulsan College of Medicine , Seoul, Korea.
authors.forename: Sung-Min
authors.initials: SM
authors.lastname: Chun
authors.affiliation: 3 Center for Cancer Genome Discovery, Asan Institute for Life Science , Seoul, Korea.
authors.forename: Deokhoon
authors.initials: D
authors.lastname: Kim
authors.affiliation: 1 Department of Internal Medicine, Asan Medical Center, University of Ulsan College of Medicine , Seoul, Korea.
authors.forename: Hyemi
authors.initials: H
authors.lastname: Kwon
authors.affiliation: 1 Department of Internal Medicine, Asan Medical Center, University of Ulsan College of Medicine , Seoul, Korea.
authors.forename: Eun Kyung
authors.initials: EK
authors.lastname: Jang
authors.affiliation: 1 Department of Internal Medicine, Asan Medical Center, University of Ulsan College of Medicine , Seoul, Korea.
authors.forename: Tae Yong
authors.initials: TY
authors.lastname: Kim
authors.affiliation: 1 Department of Internal Medicine, Asan Medical Center, University of Ulsan College of Medicine , Seoul, Korea.
authors.forename: Won Bae
authors.initials: WB
authors.lastname: Kim
authors.affiliation: 1 Department of Internal Medicine, Asan Medical Center, University of Ulsan College of Medicine , Seoul, Korea.
authors.forename: Young Kee
authors.initials: YK
authors.lastname: Shong
authors.affiliation: 2 Department of Pathology, Asan Medical Center, University of Ulsan College of Medicine , Seoul, Korea.
authors.forename: Se Jin
authors.initials: SJ
authors.lastname: Jang
authors.affiliation: 2 Department of Pathology, Asan Medical Center, University of Ulsan College of Medicine , Seoul, Korea.
authors.forename: Dong Eun
authors.initials: DE
authors.lastname: Song
authors.affiliation: 1 Department of Internal Medicine, Asan Medical Center, University of Ulsan College of Medicine , Seoul, Korea.
authors.forename: Won Gu
authors.initials: WG
authors.lastname: Kim
mesh_terms: D000368:Aged; D000369:Aged, 80 and over; D004252:DNA Mutational Analysis / Q000379:methods; D005260:Female; D005838:Genotype; D059014:High-Throughput Nucleotide Sequencing; D006801:Humans; D008297:Male; D008875:Middle Aged; D009154:Mutation; D048493:Proto-Oncogene Proteins B-raf / Q000235:genetics*; D016283:Proto-Oncogene Proteins p21(ras) / Q000235:genetics*; D065646:Thyroid Carcinoma, Anaplastic / Q000235:genetics* / Q000473:pathology; D013964:Thyroid Neoplasms / Q000235:genetics* / Q000473:pathology
chemical_list: C117307:KRAS protein, human; D048493:Proto-Oncogene Proteins B-raf; D016283:Proto-Oncogene Proteins p21(ras)