title: Targeted Transcriptional Profiling of Microdissected Biopsy Specimens Representing Early Colonic Neoplasia.
abstract: Our incomplete understanding of the critical changes that accompany the earliest stages of tumor initiation provides a substantial hurdle for the development of novel intervention strategies for cancer prevention. Premalignant lesions are inherently difficult to characterize given their diminutive size, creating technical obstacles for accurate genetic profiling. Here, we describe an approach combining laser-capture microdissection (LCM) with targeted RNA-sequencing to study the transcriptional state of epithelial and stromal cells during the earliest detectable stage of human colorectal neoplasia, the aberrant crypt foci (ACF). We provide a robust and reproducible workflow for RNA isolation, library preparation, and expression profiling of laser-captured cells from frozen OCT-embedded tissue specimens. It is anticipated that the methodological approach outlined in this report will provide a framework for a broad range of microgenomics analyses that can be routinely applied to many other premalignant tissues. J. Cell. Biochem. 117: 2677-2681, 2016. © 2016 Wiley Periodicals, Inc.
authors.affiliation: Center for Molecular Medicine, UConn Health, Farmington, Connecticut.
authors.forename: Allen
authors.initials: A
authors.lastname: Mo
authors.affiliation: Thermo Fisher Scientific, South San Francisco, California.
authors.forename: Stephen
authors.initials: S
authors.lastname: Jackson
authors.affiliation: Division of Gastroenterology, School of Medicine, UConn Health, Farmington, Connecticut.
authors.forename: Thomas J
authors.initials: TJ
authors.lastname: Devers
authors.affiliation: Center for Molecular Medicine, UConn Health, Farmington, Connecticut. rosenberg@uchc.edu.|Colon Cancer Prevention Program, Neag Comprehensive Cancer Center, UConn Health, Farmington, Connecticut. rosenberg@uchc.edu.
authors.forename: Daniel W
authors.initials: DW
authors.lastname: Rosenberg
mesh_terms: D014408:Biomarkers, Tumor / Q000235:genetics*; D003110:Colonic Neoplasms / Q000175:diagnosis* / Q000235:genetics; D055088:Early Detection of Cancer; D020869:Gene Expression Profiling; D059014:High-Throughput Nucleotide Sequencing / Q000379:methods*; D006801:Humans; D061346:Laser Capture Microdissection / Q000379:methods*; D011230:Precancerous Conditions / Q000175:diagnosis* / Q000235:genetics
chemical_list: D014408:Biomarkers, Tumor
keywords: COLONIC ABERRANT CRYPT FOCI; EARLY NEOPLASIA; LASER-CAPTURE MICRODISSECTION; TARGETED RNA-SEQ