title: Expression profiling of in vivo ductal carcinoma in situ progression models identified B cell lymphoma-9 as a molecular driver of breast cancer invasion.
abstract: BACKGROUND
  There are an estimated 60,000 new cases of ductal carcinoma in situ (DCIS) each year. A lack of understanding in DCIS pathobiology has led to overtreatment of more than half of patients. We profiled the temporal molecular changes during DCIS transition to invasive ductal carcinoma (IDC) using in vivo DCIS progression models. These studies identified B cell lymphoma-9 (BCL9) as a potential molecular driver of early invasion. BCL9 is a newly found co-activator of Wnt-stimulated β-catenin-mediated transcription. BCL9 has been shown to promote progression of multiple myeloma and colon carcinoma. However BCL9 role in breast cancer had not been previously recognized.
  
  
  METHODS
  Microarray and RNA sequencing were utilized to characterize the sequential changes in mRNA expression during DCIS invasive transition. BCL9-shRNA knockdown was performed to assess the role of BCL9 in in vivo invasion, epithelial-mesenchymal transition (EMT) and canonical Wnt-signaling. Immunofluorescence of 28 patient samples was used to assess a correlation between the expression of BCL9 and biomarkers of high risk DCIS. The cancer genome atlas data were analyzed to assess the status of BCL9 gene alterations in breast cancers.
  
  
  RESULTS
  Analysis of BCL9, by RNA and protein showed BCL9 up-regulation to be associated with DCIS transition to IDC. Analysis of patient DCIS revealed a significant correlation between high nuclear BCL9 and pathologic characteristics associated with DCIS recurrence: Estrogen receptor (ER) and progesterone receptor (PR) negative, high nuclear grade, and high human epidermal growth factor receptor2 (HER2). In vivo silencing of BCL9 resulted in the inhibition of DCIS invasion and reversal of EMT. Analysis of the TCGA data showed BCL9 to be altered in 26 % of breast cancers. This is a significant alteration when compared to HER2 (ERBB2) gene (19 %) and estrogen receptor (ESR1) gene (8 %). A significantly higher proportion of basal like invasive breast cancers compared to luminal breast cancers showed BCL9 amplification.
  
  
  CONCLUSIONS
  BCL9 is a molecular driver of DCIS invasive progression and may predispose to the development of basal like invasive breast cancers. As such, BCL9 has the potential to serve as a biomarker of high risk DCIS and as a therapeutic target for prevention of IDC.
authors.affiliation: Department of Pathology, University of Kansas Medical Center, 3901 Rainbow Blvd, Mail Stop 3003, Kansas City, KS, 66160, USA. helsarraj@kumc.edu.
authors.forename: Hanan S
authors.initials: HS
authors.lastname: Elsarraj
authors.affiliation: Department of Pathology, University of Kansas Medical Center, 3901 Rainbow Blvd, Mail Stop 3003, Kansas City, KS, 66160, USA. yhong@kumc.edu.
authors.forename: Yan
authors.initials: Y
authors.lastname: Hong
authors.affiliation: Department of Pathology, University of Kansas Medical Center, 3901 Rainbow Blvd, Mail Stop 3003, Kansas City, KS, 66160, USA. kvaldez@kumc.edu.
authors.forename: Kelli E
authors.initials: KE
authors.lastname: Valdez
authors.affiliation: School of Medicine, University of Kansas Medical Center, Kansas City, KS, 66160, USA. wmichaels@kumc.edu.
authors.forename: Whitney
authors.initials: W
authors.lastname: Michaels
authors.affiliation: School of Medicine, University of Kansas Medical Center, Kansas City, KS, 66160, USA. mhook@kumc.edu.
authors.forename: Marcus
authors.initials: M
authors.lastname: Hook
authors.affiliation: Department of Radiology, University of Kansas Medical Center, Kansas City, KS, 66160, USA. wsmith2@kumc.edu.
authors.forename: William P
authors.initials: WP
authors.lastname: Smith
authors.affiliation: Department of Cancer Biology, University of Kansas Medical Center, Kansas City, KS, 66160, USA. jchien@kumc.edu.
authors.forename: Jeremy
authors.initials: J
authors.lastname: Chien
authors.affiliation: Department of Biomedical Sciences, University at Albany-SUNY, Rensselaer, NY, 12144, USA. jherschkowitz@albany.edu.
authors.forename: Jason I
authors.initials: JI
authors.lastname: Herschkowitz
authors.affiliation: School of Public Health, University of North Carolina, Chapel Hill, NC, 27599, USA. troester@unc.edu.
authors.forename: Melissa A
authors.initials: MA
authors.lastname: Troester
authors.affiliation: Department of Chemistry, Wichita State University, Wichita, KS, 67260, USA. Moriah.Beck@wichita.edu.
authors.forename: Moriah
authors.initials: M
authors.lastname: Beck
authors.affiliation: Department of Radiology, University of Kansas Medical Center, Kansas City, KS, 66160, USA. minciardi@kumc.edu.
authors.forename: Marc
authors.initials: M
authors.lastname: Inciardi
authors.affiliation: Department of Radiology, University of Kansas Medical Center, Kansas City, KS, 66160, USA. jgatewood@kumc.edu.
authors.forename: Jason
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authors.lastname: Gatewood
authors.affiliation: School of Medicine, University of Kansas, Wichita, KS, 67214, USA. lisasmay@cox.net.
authors.forename: Lisa
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authors.lastname: May
authors.affiliation: School of Medicine, University of Kansas, Wichita, KS, 67214, USA. tcusick@wsspa.com.
authors.forename: Therese
authors.initials: T
authors.lastname: Cusick
authors.affiliation: Department of Surgery, University of Kansas Medical Center, Kansas City, KS, 66160, USA. mmcginness@kumc.edu.
authors.forename: Marilee
authors.initials: M
authors.lastname: McGinness
authors.affiliation: Department of Radiology, Truman Medical Center, Kansas City, MO, 64108, USA. lawrence.ricci@tmcmed.org.
authors.forename: Lawrence
authors.initials: L
authors.lastname: Ricci
authors.affiliation: Department of Pathology, University of Kansas Medical Center, 3901 Rainbow Blvd, Mail Stop 3003, Kansas City, KS, 66160, USA. FFAN@kumc.edu.
authors.forename: Fang
authors.initials: F
authors.lastname: Fan
authors.affiliation: Department of Pathology, University of Kansas Medical Center, 3901 Rainbow Blvd, Mail Stop 3003, Kansas City, KS, 66160, USA. OTAWFIK@kumc.edu.
authors.forename: Ossama
authors.initials: O
authors.lastname: Tawfik
authors.affiliation: Department of Surgery, Duke University, Durham, NC, 27710, USA. marks003@mc.duke.edu.
authors.forename: Jeffrey R
authors.initials: JR
authors.lastname: Marks
authors.affiliation: Kansas Intellectual and Developmental Disabilities Research Center, University of Kansas Medical Center, Kansas City, KS, 66160, USA. jknapp5@kumc.edu.
authors.forename: Jennifer R
authors.initials: JR
authors.lastname: Knapp
authors.affiliation: Department of Biostatistics, University of Kansas Medical Center, Kansas City, KS, 66160, USA. hyeh@kumc.edu.
authors.forename: Hung-Wen
authors.initials: HW
authors.lastname: Yeh
authors.affiliation: Department of Pathology, University of Kansas Medical Center, 3901 Rainbow Blvd, Mail Stop 3003, Kansas City, KS, 66160, USA. PTHOMAS@kumc.edu.
authors.forename: Patricia
authors.initials: P
authors.lastname: Thomas
authors.affiliation: Department of Pathology, Dana-Farber Cancer Institute, Harvard Medical School, Boston, MA, 02115-5450, USA. Ruben_Carrasco@dfci.harvard.edu.
authors.forename: D R
authors.initials: DR
authors.lastname: Carrasco
authors.affiliation: Department of Pathology, University of Kansas Medical Center, 3901 Rainbow Blvd, Mail Stop 3003, Kansas City, KS, 66160, USA. tfields@kumc.edu.
authors.forename: Timothy A
authors.initials: TA
authors.lastname: Fields
authors.affiliation: Department of Pathology, University of Kansas Medical Center, 3901 Rainbow Blvd, Mail Stop 3003, Kansas City, KS, 66160, USA. agodwin@kumc.edu.
authors.forename: Andrew K
authors.initials: AK
authors.lastname: Godwin
authors.affiliation: Department of Pathology, University of Kansas Medical Center, 3901 Rainbow Blvd, Mail Stop 3003, Kansas City, KS, 66160, USA. fbehbod@kumc.edu.
authors.forename: Fariba
authors.initials: F
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mesh_terms: D000818:Animals; D014408:Biomarkers, Tumor / Q000235:genetics; D001943:Breast Neoplasms / Q000235:genetics* / Q000473:pathology*; D018270:Carcinoma, Ductal, Breast / Q000235:genetics* / Q000473:pathology; D002285:Carcinoma, Intraductal, Noninfiltrating / Q000235:genetics* / Q000473:pathology*; D018450:Disease Progression; D058750:Epithelial-Mesenchymal Transition / Q000235:genetics; D005260:Female; D006801:Humans; D051379:Mice; D016688:Mice, Inbred NOD; D016513:Mice, SCID; D009363:Neoplasm Proteins / Q000235:genetics*; D009364:Neoplasm Recurrence, Local / Q000235:genetics / Q000473:pathology; D018719:Erb-b2 Receptor Tyrosine Kinases / Q000235:genetics; D011960:Receptors, Estrogen / Q000235:genetics; D011980:Receptors, Progesterone / Q000235:genetics; D014157:Transcription Factors; D014158:Transcription, Genetic / Q000235:genetics; D059467:Transcriptome / Q000235:genetics*; D015854:Up-Regulation / Q000235:genetics; D051153:Wnt Proteins / Q000235:genetics; D051176:beta Catenin / Q000235:genetics
chemical_list: D014408:Biomarkers, Tumor; D009363:Neoplasm Proteins; D018719:Erb-b2 Receptor Tyrosine Kinases; D011960:Receptors, Estrogen; D011980:Receptors, Progesterone; D014157:Transcription Factors; D051153:Wnt Proteins; D051176:beta Catenin; C111180:BCL9 protein, human; C508053:ERBB2 protein, human