Vitamin D Regulation of the Uridine Phosphorylase 1 Gene and Uridine-Induced DNA Damage in Colon in African Americans and European Americans

• Published in: Gastroenterology (New York, N.Y. 1943) Vol. 155; no. 4; pp. 1192 - 1204.e9
• Main Authors: Bhasin, Nobel, Alleyne, Dereck, Gray, Olivia A., Kupfer, Sonia S.
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 01.10.2018
• Subjects: African Americans - genetics; Animals; Binding Sites; Calcitriol - pharmacology; Cell Line; Colon - drug effects; Colon - enzymology; Colon - pathology; CRC; Cytoprotection; DNA Damage - drug effects; Epithelial Cells - drug effects; Epithelial Cells - enzymology; Epithelial Cells - pathology; Ethnicity; European Continental Ancestry Group - genetics; Gene Expression Regulation, Enzymologic; Genetics; Humans; Mice; Organoids - drug effects; Organoids - enzymology; Organoids - pathology; Polymorphism, Genetic; Promoter Regions, Genetic; Receptors, Calcitriol - agonists; Receptors, Calcitriol - genetics; Receptors, Calcitriol - metabolism; Risk Factor; Time Factors; Tissue Culture Techniques; Uridine - metabolism; Uridine - toxicity; Uridine Phosphorylase - genetics; Uridine Phosphorylase - metabolism; GTEx; dUTP; LASAGNA; VDR; DAPI; mRNA; siRNA; eQTL; ChIP; bp; Ethnicity; FLARE; SNP; CRC; TSS; kb; YAMC; Genetics; ATM; UPP1; Risk Factor; PCR; 5-FU; ATR
• ISSN: 0016-5085; 1528-0012
• DOI: 10.1053/j.gastro.2018.06.049

African Americans have the greatest colorectal cancer (CRC) burden in the United States; interethnic differences in protective effects of vitamin D might contribute to disparities. 1α,25(OH)2D3 vitamin D (the active form of vitamin D) induces transcription of the uridine phosphorylase gene (UPP1) in colon tissues of European Americans but to a lesser extent in colon tissues of African Americans. UPP1-knockout mice have increased intestinal concentrations of uridine and Deoxyuridine triphosphate (dUTP), have increased uridine-induced DNA damage, and develop colon tumors. We studied 1α,25(OH)2D3 regulation of UPP1 and uridine-induced DNA damage in the colon and differences in these processes between African and European Americans. We quantified expression and activity of UPP1 in response to 1α,25(OH)2D3 in young adult mouse colonic cells, human CRC cells (LS174T), and organoids (derived from rectosigmoid biopsy samples of healthy individuals undergoing colonoscopies) using quantitative polymerase chain reaction, immunoblot, and immunocytochemistry assays. Binding of the vitamin D receptor to UPP1 was tested by chromatin immunoprecipitation. Uridine-induced DNA damage was measured by fragment-length analysis in repair enzyme assays. Allele-specific 1α,25(OH)2D3 responses were tested using luciferase assays. Vitamin D increased levels of UPP1 mRNA, protein, and enzymatic activity and increased vitamin D receptor binding to the UPP1 promoter in young adult mouse colonic cells, LS174T cells, and organoids. 1α,25(OH)2D3 significantly reduced levels of uridine and uridine-induced DNA damage in these cells, which required UPP1 expression. Organoids derived from colon tissues of African Americans expressed lower levels of UPP1 after exposure to 1α,25(OH)2D3 and had increased uridine-induced DNA damage compared with organoids derived from tissues of European Americans. Luciferase assays with the T allele of single nucleotide polymorphism rs28605337 near UPP1, which is found more frequently in African Americans than European Americans, expressed lower levels of UPP1 after exposure to 1α,25(OH)2D3 than assays without this variant. We found vitamin D to increase expression of UPP1, leading to reduce uridine-induced DNA damage, in colon cells and organoids. A polymorphism in UPP1 found more frequently in African Americans than European Americans reduced UPP1 expression upon cell exposure to 1α,25(OH)2D3. Differences in expression of UPP1 in response to vitamin D could contribute to the increased risk of CRC in African Americans.