title: Development of a colon cancer GEMM-derived orthotopic transplant model for drug discovery and validation.
abstract: OBJECTIVE
  Effective therapies for KRAS-mutant colorectal cancer (CRC) are a critical unmet clinical need. Previously, we described genetically engineered mouse models (GEMM) for sporadic Kras-mutant and non-mutant CRC suitable for preclinical evaluation of experimental therapeutics. To accelerate drug discovery and validation, we sought to derive low-passage cell lines from GEMM Kras-mutant and wild-type tumors for in vitro screening and transplantation into the native colonic environment of immunocompetent mice for in vivo validation.
  
  
  METHODS
  Cell lines were derived from Kras-mutant and non-mutant GEMM tumors under defined media conditions. Growth kinetics, phosphoproteomes, transcriptomes, drug sensitivity, and metabolism were examined. Cell lines were implanted in mice and monitored for in vivo tumor analysis.
  
  
  RESULTS
  Kras-mutant cell lines displayed increased proliferation, mitogen-activated protein kinase signaling, and phosphoinositide-3 kinase signaling. Microarray analysis identified significant overlap with human CRC-related gene signatures, including KRAS-mutant and metastatic CRC. Further analyses revealed enrichment for numerous disease-relevant biologic pathways, including glucose metabolism. Functional assessment in vitro and in vivo validated this finding and highlighted the dependence of Kras-mutant CRC on oncogenic signaling and on aerobic glycolysis.
  
  
  CONCLUSIONS
  We have successfully characterized a novel GEMM-derived orthotopic transplant model of human KRAS-mutant CRC. This approach combines in vitro screening capability using low-passage cell lines that recapitulate human CRC and potential for rapid in vivo validation using cell line-derived tumors that develop in the colonic microenvironment of immunocompetent animals. Taken together, this platform is a clear advancement in preclinical CRC models for comprehensive drug discovery and validation efforts.
authors: Dana-Farber Cancer Institute, Boston, Massachusetts, USA. Eric.Martin@pfizer.com
authors: Eric S
authors: ES
authors: Martin
authors: Peter J
authors: PJ
authors: Belmont
authors: Mark J
authors: MJ
authors: Sinnamon
authors: Larissa Georgeon
authors: LG
authors: Richard
authors: Jing
authors: J
authors: Yuan
authors: Erin M
authors: EM
authors: Coffee
authors: Jatin
authors: J
authors: Roper
authors: Lydia
authors: L
authors: Lee
authors: Pedram
authors: P
authors: Heidari
authors: Sophia Y
authors: SY
authors: Lunt
authors: Gautam
authors: G
authors: Goel
authors: Xiadong
authors: X
authors: Ji
authors: Zhi
authors: Z
authors: Xie
authors: Tao
authors: T
authors: Xie
authors: John
authors: J
authors: Lamb
authors: Scott L
authors: SL
authors: Weinrich
authors: Todd
authors: T
authors: VanArsdale
authors: Roderick T
authors: RT
authors: Bronson
authors: Ramnik J
authors: RJ
authors: Xavier
authors: Matthew G
authors: MG
authors: Vander Heiden
authors: Julie L C
authors: JL
authors: Kan
authors: Umar
authors: U
authors: Mahmood
authors: Kenneth E
authors: KE
authors: Hung
mesh_terms: D000818:Animals; D063646:Carcinogenesis / Q000235:genetics / Q000378:metabolism; D045744:Cell Line, Tumor; D016000:Cluster Analysis; D003110:Colonic Neoplasms / Q000188:drug therapy / Q000235:genetics* / Q000378:metabolism; D004195:Disease Models, Animal; D020869:Gene Expression Profiling; D017491:Genes, APC; D016158:Genes, p53; D005838:Genotype; D005947:Glucose / Q000378:metabolism; D006801:Humans; D019344:Lactic Acid / Q000096:biosynthesis; D051379:Mice; D008822:Mice, Transgenic; D020928:Mitogen-Activated Protein Kinases / Q000378:metabolism; D009154:Mutation; D019869:Phosphatidylinositol 3-Kinases / Q000378:metabolism; D015398:Signal Transduction; D058570:TOR Serine-Threonine Kinases / Q000378:metabolism; D060449:Wnt Signaling Pathway; D023041:Xenograft Model Antitumor Assays; D018631:ras Proteins / Q000235:genetics* / Q000378:metabolism
chemical_list: D005947:Glucose; D019344:Lactic Acid; D020928:Mitogen-Activated Protein Kinases; D019869:Phosphatidylinositol 3-Kinases; D058570:TOR Serine-Threonine Kinases; D018631:ras Proteins
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