DNA methylome profiling identifies novel methylated genes in African American patients with colorectal neoplasia.
The identification of genes that are differentially methylated in colorectal cancer (CRC) has potential value for both diagnostic and therapeutic interventions specifically in high-risk populations such as African Americans (AAs). However, DNA methylation patterns in CRC, especially in AAs, have not been systematically explored and remain poorly understood. Here, we performed DNA methylome profiling to identify the methylation status of CpG islands within candidate genes involved in critical pathways important in the initiation and development of CRC. We used reduced representation bisulfite sequencing (RRBS) in colorectal cancer and adenoma tissues that were compared with DNA methylome from a healthy AA subject's colon tissue and peripheral blood DNA. The identified methylation markers were validated in fresh frozen CRC tissues and corresponding normal tissues from AA patients diagnosed with CRC at Howard University Hospital. We identified and validated the methylation status of 355 CpG sites located within 16 gene promoter regions associated with CpG islands. Fifty CpG sites located within CpG islands-in genes ATXN7L1 (2), BMP3 (7), EID3 (15), GAS7 (1), GPR75 (24), and TNFAIP2 (1)-were significantly hypermethylated in tumor vs. normal tissues (P<0.05). The methylation status of BMP3, EID3, GAS7, and GPR75 was confirmed in an independent, validation cohort. Ingenuity pathway analysis mapped three of these markers (GAS7, BMP3 and GPR) in the insulin and TGF-β1 network-the two key pathways in CRC. In addition to hypermethylated genes, our analysis also revealed that LINE-1 repeat elements were progressively hypomethylated in the normal-adenoma-cancer sequence. We conclude that DNA methylome profiling based on RRBS is an effective method for screening aberrantly methylated genes in CRC. While previous studies focused on the limited identification of hypermethylated genes, ours is the first study to systematically and comprehensively identify novel hypermethylated genes, as well as hypomethylated LINE-1 sequences, which may serve as potential biomarkers for CRC in African Americans. Our discovered biomarkers were intimately linked to the insulin/TGF-B1 pathway, further strengthening the association of diabetic disorders with colon oncogenic transformation.
Department of Medicine and Cancer Center; Department of Pathology; Howard University College of Medicine; Washington DC, USA.
Hassan
H
Ashktorab
Department of Medicine and Cancer Center; Department of Pathology; Howard University College of Medicine; Washington DC, USA.
M
M
Daremipouran
Department of Medicine and Cancer Center; Department of Pathology; Howard University College of Medicine; Washington DC, USA; Baylor Research Institute and Charles A Sammons Cancer Center; Baylor University Medical Center, Dallas, TX USA.
Ajay
A
Goel
Department of Medicine and Cancer Center; Department of Pathology; Howard University College of Medicine; Washington DC, USA; Hithru; Laurel, MD USA.
Sudhir
S
Varma
Department of Medicine and Cancer Center; Department of Pathology; Howard University College of Medicine; Washington DC, USA; Zymo Research Corp.; Irvine, CA USA.
R
R
Leavitt
Zymo Research Corp.; Irvine, CA USA.
Xueguang
X
Sun
Department of Medicine and Cancer Center; Department of Pathology; Howard University College of Medicine; Washington DC, USA.
Hassan
H
Brim
D000236:Adenoma / Q000235:genetics / Q000378:metabolism*; D001741:Black or African American; D014408:Biomarkers, Tumor / Q000235:genetics / Q000378:metabolism; D016022:Case-Control Studies; D015179:Colorectal Neoplasms / Q000235:genetics / Q000378:metabolism*; D018899:CpG Islands; D019175:DNA Methylation; D015894:Genome, Human; D006801:Humans; D020084:Long Interspersed Nucleotide Elements; D011401:Promoter Regions, Genetic; D017422:Sequence Analysis, DNA
D014408:Biomarkers, Tumor
African Americans; RRBS; colon cancer; neoplasia