Biomarkers of High Grade Dysplasia in Barrett's Esophagus

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Methods (1)• Esophageal biopsies (11 cases)– Formalin-fixed, paraffin-embedded. 2004-7– Inclusion criteria• Good tissue orientation• Easily defined areas of HGD and normal BE (SAME PATIENT!!)• HGD confirmed by 2-3 GI pathologists• Must have high quality mRNA–10μm sections deparaffinized, scraped– RNA extracted with Paradise kit (Arcturus)– DNA removed (DNAse)– RNA quality determined by» Agilent 2100 Bioanalyser» 5’ and 3’ β-actin RT-PCR (oligo-dT)–3’:5’β -actin < 10; >15ng
Specimen Evaluation40 cases (1-6 blocks of biopsies per case) received fromcollaborators26 had both NBE and HGD available for LCM11 had sufficient quantities of RNA and passed 3’5’ QCevaluation for intact RNA
Page 1: Biomarkers of High GradeDysplasia i
Page 7: Barrett’s Esophagus• 5-15% of a
Page 12 and 13: Barrett’s Esophagus SequelaeUlcer
Page 14 and 15: Histopathologic Dx of DysplasiaDysp
Page 16 and 17: Dysplasia in Barrett’s Esophagus
Page 18: LGD
Page 21 and 22: Dysplasia in BEPitfalls• Sampling
Page 23 and 24: Ancillary Techniques in theScreenin
Page 25: Advantages over other microarray st
Page 29 and 30: Biopsy of Normal BE
Page 31 and 32: Use of Aperio ScanScope for Remote
Page 33 and 34: BioinformaticsExpression signals no
Page 35: Dysregulated Genes in HGD by Functi
Page 38 and 39: CAMK2N1MUC5ACHSD17BPGDHAMEP1AALDOBA
Page 40 and 41: Topoisomerase II alpha
Page 42 and 43: S100-A9
Page 44 and 45: Immunohistochemical stains for topo
Page 46 and 47: Thanks to…….Patricia A. Meitner

Biomarkers of High Grade Dysplasia in Barrett's Esophagus

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