title: Establishment and characterization of novel human primary and metastatic anaplastic thyroid cancer cell lines and their genomic evolution over a year as a primagraft.
abstract: BACKGROUND
  Anaplastic thyroid cancer (ATC) has no effective treatment, resulting in a high rate of mortality. We established cell lines from a primary ATC and its lymph node metastasis, and investigated the molecular factors and genomic changes associated with tumor growth.
  
  
  OBJECTIVE
  The aim of the study was to understand the molecular and genomic changes of highly aggressive ATC and its clonal evolution to develop rational therapies.
  
  
  METHODS
  We established unique cell lines from primary (OGK-P) and metastatic (OGK-M) ATC specimen, as well as primagraft from the metastatic ATC, which was serially xeno-transplanted for more than 1 year in NOD scid gamma mice were established. These cell lines and primagraft were used as tools to examine gene expression, copy number changes, and somatic mutations using RNA array, SNP Chip, and whole exome sequencing.
  
  
  RESULTS
  Mice carrying sc (OGK-P and OGK-M) tumors developed splenomegaly and neutrophilia with high expression of cytokines including CSF1, CSF2, CSF3, IL-1β, and IL-6. Levels of HIF-1α and its targeted genes were also elevated in these tumors. The treatment of tumor carrying mice with Bevacizumab effectively decreased tumor growth, macrophage infiltration, and peripheral WBCs. SNP chip analysis showed homozygous deletion of exons 3-22 of the PARD3 gene in the cells. Forced expression of PARD3 decreased cell proliferation, motility, and invasiveness, restores cell-cell contacts and enhanced cell adhesion. Next generation exome sequencing identified the somatic changes present in the primary, metastatic, and primagraft tumors demonstrating evolution of the mutational signature over the year of passage in vivo.
  
  
  CONCLUSIONS
  To our knowledge, we established the first paired human primary and metastatic ATC cell lines offering unique possibilities for comparative functional investigations in vitro and in vivo. Our exome sequencing also identified novel mutations, as well as clonal evolution in both the metastasis and primagraft.
authors.affiliation: Cancer Science Institute (CSI) of Singapore (M.G., D.K., S.V., A.M.T., L.-z.L., K.T.T., W.J.C., H.Y., H.P.K.), National University of Singapore, Singapore 117599, Singapore; Division of Hematology/Oncology (R.O., Q.H., T.A., S.G., H.P.K.), Cedars-Sinai Medical Center, Los Angeles, California 90048; Graduate School of Medicine (Y.N., S.O.), The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-8655, Japan; Department of Medicine (G.D.B.), Cedars-Sinai Medical Center, Los Angeles, California 90048; Department of Pathology and Laboratory Medicine (J.W.S., N.B.D.), David Geffen School of Medicine, Los Angeles, California 90048; and National University Cancer Institute (H.P.K.), National University Hospital, Singapore 117599.
authors.forename: Manoj
authors.initials: M
authors.lastname: Garg
authors.forename: Ryoko
authors.initials: R
authors.lastname: Okamoto
authors.forename: Yasunobu
authors.initials: Y
authors.lastname: Nagata
authors.forename: Deepika
authors.initials: D
authors.lastname: Kanojia
authors.forename: Subhashree
authors.initials: S
authors.lastname: Venkatesan
authors.forename: Anand
authors.initials: A
authors.lastname: M T
authors.forename: Glenn D
authors.initials: GD
authors.lastname: Braunstein
authors.forename: Jonathan W
authors.initials: JW
authors.lastname: Said
authors.forename: Ngan B
authors.initials: NB
authors.lastname: Doan
authors.forename: Quoc
authors.initials: Q
authors.lastname: Ho
authors.forename: Tadayuki
authors.initials: T
authors.lastname: Akagi
authors.forename: Sigal
authors.initials: S
authors.lastname: Gery
authors.forename: Li-Zhen
authors.initials: LZ
authors.lastname: Liu
authors.forename: Kar Tong
authors.initials: KT
authors.lastname: Tan
authors.forename: Wee Joo
authors.initials: WJ
authors.lastname: Chng
authors.forename: Henry
authors.initials: H
authors.lastname: Yang
authors.forename: Seishi
authors.initials: S
authors.lastname: Ogawa
authors.forename: H Phillip
authors.initials: HP
authors.lastname: Koeffler
mesh_terms: D000368:Aged; D000818:Animals; D045744:Cell Line, Tumor / Q000473:pathology*; D049109:Cell Proliferation; D006801:Humans; D008297:Male; D051379:Mice; D016688:Mice, Inbred NOD; D009368:Neoplasm Transplantation; D009504:Neutrophils / Q000473:pathology; D013163:Splenomegaly / Q000209:etiology / Q000473:pathology; D065646:Thyroid Carcinoma, Anaplastic / Q000150:complications / Q000473:pathology*; D013964:Thyroid Neoplasms / Q000150:complications / Q000473:pathology*