title: Laser capture microdissection-reduced representation bisulfite sequencing (LCM-RRBS) maps changes in DNA methylation associated with gonadectomy-induced adrenocortical neoplasia in the mouse.
abstract: DNA methylation is a mechanism for long-term transcriptional regulation and is required for normal cellular differentiation. Failure to properly establish or maintain DNA methylation patterns leads to cell dysfunction and diseases such as cancer. Identifying DNA methylation signatures in complex tissues can be challenging owing to inaccurate cell enrichment methods and low DNA yields. We have developed a technique called laser capture microdissection-reduced representation bisulfite sequencing (LCM-RRBS) for the multiplexed interrogation of the DNA methylation status of cytosine-guanine dinucleotide islands and promoters. LCM-RRBS accurately and reproducibly profiles genome-wide methylation of DNA extracted from microdissected fresh frozen or formalin-fixed paraffin-embedded tissue samples. To demonstrate the utility of LCM-RRBS, we characterized changes in DNA methylation associated with gonadectomy-induced adrenocortical neoplasia in the mouse. Compared with adjacent normal tissue, the adrenocortical tumors showed reproducible gains and losses of DNA methylation at genes involved in cell differentiation and organ development. LCM-RRBS is a rapid, cost-effective, and sensitive technique for analyzing DNA methylation in heterogeneous tissues and will facilitate the investigation of DNA methylation in cancer and organ development.
authors.affiliation: Department of Genetics, Center for Genome Sciences, Washington University School of Medicine, 4444 Forest Park Parkway, St. Louis, Missouri 63110, USA.
authors.forename: Maximiliaan
authors.initials: M
authors.lastname: Schillebeeckx
authors.forename: Anja
authors.initials: A
authors.lastname: Schrade
authors.forename: Ann-Kathrin
authors.initials: AK
authors.lastname: Löbs
authors.forename: Marjut
authors.initials: M
authors.lastname: Pihlajoki
authors.forename: David B
authors.initials: DB
authors.lastname: Wilson
authors.forename: Robi D
authors.initials: RD
authors.lastname: Mitra
mesh_terms: D000310:Adrenal Gland Neoplasms / Q000209:etiology / Q000235:genetics*; D000818:Animals; D002369:Castration; D019175:DNA Methylation; D006801:Humans; D061346:Laser Capture Microdissection; D051379:Mice; D016133:Polymerase Chain Reaction; D017422:Sequence Analysis, DNA; D013447:Sulfites
chemical_list: D013447:Sulfites; C042345:hydrogen sulfite