GENOME-WIDE METHYLOME ANALYSIS USING METHYLCAP-SEQ UNCOVERS NOVEL METHYLATED MARKERS FOR CANINE LYMPHOMA CELL LINE CLBL-1

Abstract

The most common type of tumor occurring in canine is the lymphomas. Epigenetic factors like DNA methylation is one of the important factors for causing cancer in canine. Cytology-based screening methods, such as fine needle aspiration (FNA) or immunocytochemistry (ICC), for canine lymphomas lack sensitivity. Despite an improvement in the detection of canine lymphomas by the DNA methylation analysis of lymph scrapings, only few methylation markers have been described. Even though technique like Digital Restriction Enzyme Analysis of Methylation (DREAM) can analyze the methylation profile of canine lymphoma cell line CLBL-1 at very low experimental cost without specific antibodies, the sensitivity and accuracy remain low in comparison to MethylCap-seq. In this study, a methylated DNA fragment of CLBL-1 was successfully captured by a specific protein for MethylCap-seq, and the methylation map of CLBL-1 was reconstructed by high throughput sequencing. For the first time, the genomic methylation region of canine CLBL-1 was analyzed. Using CLBL-1, we aimed to identify novel methylated markers for monitoring canine lymphomas. With the aid of methylation maps, several researchers have attempted to explore the relationship between DNA methylation and specific cancers. Additionally, the discovery of important biomarkers can predict or observe the course of canine cancer. Finally, the proposed advanced methylation information is likely to be suitable as a useful reference for clinical application of canine lymphoma research.