title: A gene expression signature of epithelial tubulogenesis and a role for ASPM in pancreatic tumor progression.
abstract: OBJECTIVE
  Many patients with pancreatic ductal adenocarcinoma (PDAC) develop recurrent or metastatic diseases after surgery, so it is important to identify those most likely to benefit from aggressive therapy. Disruption of tissue microarchitecture is an early step in pancreatic tumorigenesis and a parameter used in pathology grading of glandular tumors. We investigated whether changes in gene expression during pancreatic epithelial morphogenesis were associated with outcomes of patients with PDAC after surgery.
  
  
  METHODS
  We generated architectures of human pancreatic duct epithelial cells in a 3-dimensional basement membrane matrix. We identified gene expression profiles of the cells during different stages of tubular morphogenesis (tubulogenesis) and of PANC-1 cells during spheroid formation. Differential expression of genes was confirmed by immunoblot analysis. We compared the gene expression profile associated with pancreatic epithelial tubulogenesis with that of PDAC samples from 27 patients, as well as with their outcomes after surgery.
  
  
  RESULTS
  We identified a gene expression profile associated with tubulogenesis that resembled the profile of human pancreatic tissue with differentiated morphology and exocrine function. Patients with PDACs with this profile fared well after surgery. Based on this profile, we established a 6-28 gene tubulogenesis-specific signature that accurately determined the prognosis of independent cohorts of patients with PDAC (total n = 128; accuracy = 81.2%-95.0%). One gene, ASPM, was down-regulated during tubulogenesis but up-regulated in human PDAC cell lines and tumor samples; up-regulation correlated with patient outcomes (Cox regression P = .0028). Bioinformatic, genetic, biochemical, functional, and clinical correlative studies showed that ASPM promotes aggressiveness of PDAC by maintaining Wnt-β-catenin signaling and stem cell features of PDAC cells.
  
  
  CONCLUSIONS
  We identified a gene expression profile associated with pancreatic epithelial tubulogenesis and a tissue architecture-specific signature of PDAC cells that is associated with patient outcomes after surgery.
authors.affiliation: Laboratory for Tumor Epigenetics and Stemness, National Institute of Cancer Research and Translational Center for Glandular Malignancies, National Health Research Institutes, Tainan, Taiwan; Department of Pathology, National Cheng-Kung University Hospital and College of Medicine, National Cheng Kung University, Tainan, Taiwan.
authors.forename: Wei-Yu
authors.initials: WY
authors.lastname: Wang
authors.forename: Chung-Chi
authors.initials: CC
authors.lastname: Hsu
authors.forename: Ting-Yun
authors.initials: TY
authors.lastname: Wang
authors.forename: Chi-Rong
authors.initials: CR
authors.lastname: Li
authors.forename: Ya-Chin
authors.initials: YC
authors.lastname: Hou
authors.forename: Jui-Mei
authors.initials: JM
authors.lastname: Chu
authors.forename: Chung-Ta
authors.initials: CT
authors.lastname: Lee
authors.forename: Ming-Sheng
authors.initials: MS
authors.lastname: Liu
authors.forename: Jimmy J-M
authors.initials: JJ
authors.lastname: Su
authors.forename: Kuan-Ying
authors.initials: KY
authors.lastname: Jian
authors.forename: Shenq-Shyang
authors.initials: SS
authors.lastname: Huang
authors.forename: Shih-Sheng
authors.initials: SS
authors.lastname: Jiang
authors.forename: Yan-Shen
authors.initials: YS
authors.lastname: Shan
authors.forename: Pin-Wen
authors.initials: PW
authors.lastname: Lin
authors.forename: Yin-Ying
authors.initials: YY
authors.lastname: Shen
authors.forename: Michael T-L
authors.initials: MT
authors.lastname: Lee
authors.forename: Tze-Sian
authors.initials: TS
authors.lastname: Chan
authors.forename: Chun-Chao
authors.initials: CC
authors.lastname: Chang
authors.forename: Chung-Hsing
authors.initials: CH
authors.lastname: Chen
authors.forename: I-Shou
authors.initials: IS
authors.lastname: Chang
authors.forename: Yen-Ling
authors.initials: YL
authors.lastname: Lee
authors.forename: Li-Tzong
authors.initials: LT
authors.lastname: Chen
authors.forename: Kelvin K
authors.initials: KK
authors.lastname: Tsai
mesh_terms: D000818:Animals; D014408:Biomarkers, Tumor / Q000235:genetics / Q000502:physiology; D021441:Carcinoma, Pancreatic Ductal / Q000235:genetics / Q000473:pathology*; D002454:Cell Differentiation / Q000235:genetics* / Q000502:physiology; D002465:Cell Movement / Q000235:genetics / Q000502:physiology; D004195:Disease Models, Animal; D018450:Disease Progression; D004848:Epithelium / Q000473:pathology; D005500:Follow-Up Studies; D015972:Gene Expression Regulation, Neoplastic / Q000235:genetics* / Q000502:physiology; D064593:Heterografts; D006801:Humans; D051379:Mice; D016688:Mice, Inbred NOD; D016513:Mice, SCID; D009419:Nerve Tissue Proteins / Q000235:genetics / Q000502:physiology*; D010183:Pancreatic Ducts / Q000473:pathology*; D010190:Pancreatic Neoplasms / Q000235:genetics / Q000473:pathology*; D011379:Prognosis; D012189:Retrospective Studies; D015398:Signal Transduction / Q000235:genetics / Q000502:physiology; D059467:Transcriptome / Q000235:genetics* / Q000502:physiology; D051153:Wnt Proteins / Q000502:physiology; D051176:beta Catenin / Q000502:physiology
chemical_list: C467202:ASPM protein, human; D014408:Biomarkers, Tumor; D009419:Nerve Tissue Proteins; D051153:Wnt Proteins; D051176:beta Catenin
keywords: 3-dimensional; 3D; ASPM; Biomarker; C-index; CSC; Dvl; HPDE; PDAC; Pancreatic Cancer; Prognostic Factor; Tumor Progression; abnormal spindle-like microcephaly associated; cancer stem cell; concordance index; dishevelled; human pancreatic ductal epithelial; pancreatic ductal adenocarcinoma; rBM; reconstituted basement membrane; shRNA; short-hairpin RNA