PMID- 34480209 OWN - NLM STAT- MEDLINE DCOM- 20220106 LR - 20220106 IS - 1435-5922 (Electronic) IS - 0944-1174 (Linking) VI - 56 IP - 11 DP - 2021 Nov TI - Increased DNA-incorporated thiopurine metabolite as a possible mechanism for leukocytopenia through cell apoptosis in inflammatory bowel disease patients with NUDT15 mutation. PG - 999-1007 LID - 10.1007/s00535-021-01820-0 [doi] AB - BACKGROUND AND AIMS: Polymorphisms in the nucleotide diphosphate-linked moiety X-type motif 15 (NUDT15) gene are associated with thiopurine-induced leukopenia in patients with inflammatory bowel disease (IBD). NUDT15-associated subcellular thiopurine metabolism has not been investigated in primary lymphocytes. We hypothesized that NUDT15 mutation increases DNA-incorporated deoxythioguanosine (dTG) and induces apoptosis in lymphocytes. METHODS: DNA-incorporated dTG in peripheral blood mononuclear cells (PBMCs) and 6-thioguanine nucleotides (6-TGN) in red blood cells were measured in patients with IBD undergoing thiopurine treatment. The association of a single nucleotide polymorphism for NUDT15 (rs116855232) with dTG(PBMC) was examined. The pro-apoptotic effect of DNA-incorporated dTG was examined ex vivo in association with NUDT15 genotypes by co-culturing patient-derived peripheral CD4(+) T lymphocytes with 6-thioguanine (6-TG). RESULTS: dTG(PBMC) was significantly higher in NUDT15 variants than in non-variants. dTG(PBMC), but not 6-TGN(RBC), negatively correlated with peripheral lymphocyte counts (r = - 0.31 and - 0.12, p = 0.012 and 0.173, respectively). DNA-incorporated dTG significantly accumulated to a greater extent in lymphocytes from NUDT15 variants when co-cultured with 6-TG ex vivo than in those from non-variants and was associated with decreased proliferation and increased apoptosis. CONCLUSION: Increased DNA-incorporated dTG may be responsible for thiopurine-induced leukocytopenia through cell apoptosis in IBD patients with NUDT15 mutation. CI - (c) 2021. Japanese Society of Gastroenterology. FAU - Toyonaga, Takahiko AU - Toyonaga T AD - Center for Advanced IBD Research and Treatment, Kitasato University Kitasato Institute Hospital, 5-9-1 Shirokane, Minato-ku, Tokyo, 108-8642, Japan. AD - Department of Gastroenterology, Kitasato University Kitasato Institute Hospital, 5-9-1 Shirokane, Minato-ku, Tokyo, 108-8642, Japan. FAU - Kobayashi, Taku AU - Kobayashi T AUID- ORCID: 0000-0002-2073-4234 AD - Center for Advanced IBD Research and Treatment, Kitasato University Kitasato Institute Hospital, 5-9-1 Shirokane, Minato-ku, Tokyo, 108-8642, Japan. kobataku@insti.kitasato-u.ac.jp. FAU - Kuronuma, Satoshi AU - Kuronuma S AD - Center for Advanced IBD Research and Treatment, Kitasato University Kitasato Institute Hospital, 5-9-1 Shirokane, Minato-ku, Tokyo, 108-8642, Japan. AD - Biomedical Laboratory, Department of Research, Kitasato University Kitasato Institute Hospital, 5-9-1 Shirokane, Minato-ku, Tokyo, 108-8642, Japan. FAU - Ueno, Aito AU - Ueno A AD - Center for Advanced IBD Research and Treatment, Kitasato University Kitasato Institute Hospital, 5-9-1 Shirokane, Minato-ku, Tokyo, 108-8642, Japan. FAU - Kiyohara, Hiroki AU - Kiyohara H AD - Department of Gastroenterology, Kitasato University Kitasato Institute Hospital, 5-9-1 Shirokane, Minato-ku, Tokyo, 108-8642, Japan. FAU - Okabayashi, Shinji AU - Okabayashi S AD - Center for Advanced IBD Research and Treatment, Kitasato University Kitasato Institute Hospital, 5-9-1 Shirokane, Minato-ku, Tokyo, 108-8642, Japan. FAU - Takeuchi, Osamu AU - Takeuchi O AD - Biomedical Laboratory, Department of Research, Kitasato University Kitasato Institute Hospital, 5-9-1 Shirokane, Minato-ku, Tokyo, 108-8642, Japan. FAU - Redfern, Christopher P F AU - Redfern CPF AD - The Newcastle Cancer Centre, Northern Institute for Cancer Research, Newcastle University, Newcastle upon Tyne, NE2 4HH, UK. FAU - Terai, Hideki AU - Terai H AD - Department of Respiratory Medicine, Kitasato University Kitasato Institute Hospital, 5-9-1 Shirokane, Minato-ku, Tokyo, 108-8642, Japan. FAU - Ozaki, Ryo AU - Ozaki R AD - Department of Gastroenterology, Kitasato University Kitasato Institute Hospital, 5-9-1 Shirokane, Minato-ku, Tokyo, 108-8642, Japan. FAU - Sagami, Shintaro AU - Sagami S AD - Center for Advanced IBD Research and Treatment, Kitasato University Kitasato Institute Hospital, 5-9-1 Shirokane, Minato-ku, Tokyo, 108-8642, Japan. AD - Department of Gastroenterology, Kitasato University Kitasato Institute Hospital, 5-9-1 Shirokane, Minato-ku, Tokyo, 108-8642, Japan. FAU - Nakano, Masaru AU - Nakano M AD - Department of Gastroenterology, Kitasato University Kitasato Institute Hospital, 5-9-1 Shirokane, Minato-ku, Tokyo, 108-8642, Japan. FAU - Coulthard, Sally A AU - Coulthard SA AD - The Newcastle Cancer Centre, Northern Institute for Cancer Research, Newcastle University, Newcastle upon Tyne, NE2 4HH, UK. FAU - Tanaka, Yoichi AU - Tanaka Y AD - Department of Clinical Pharmacy, Center for Clinical Pharmacy and Sciences, School of Pharmacy, Kitasato University, 5-9-1, Shirokane, Minato-ku, Tokyo, 108-8641, Japan. FAU - Hibi, Toshifumi AU - Hibi T AD - Center for Advanced IBD Research and Treatment, Kitasato University Kitasato Institute Hospital, 5-9-1 Shirokane, Minato-ku, Tokyo, 108-8642, Japan. LA - eng PT - Journal Article DEP - 20210904 PL - Japan TA - J Gastroenterol JT - Journal of gastroenterology JID - 9430794 RN - EC 2.1.1.- (Methyltransferases) RN - EC 2.1.1.67 (thiopurine methyltransferase) RN - EC 2.6.1.- (NUDT15 protein, human) RN - EC 3.6.1.- (Pyrophosphatases) SB - IM MH - Adult MH - Apoptosis MH - Cross-Sectional Studies MH - Female MH - Humans MH - Inflammatory Bowel Diseases/blood/*complications MH - Japan MH - Leukopenia/blood/*etiology MH - Male MH - Methyltransferases/*adverse effects/analysis MH - Middle Aged MH - Pyrophosphatases/*analysis/blood OTO - NOTNLM OT - Inflammatory bowel disease OT - NUDT15 OT - Thiopurine EDAT- 2021/09/05 06:00 MHDA- 2022/01/07 06:00 CRDT- 2021/09/04 06:10 PHST- 2021/05/25 00:00 [received] PHST- 2021/08/21 00:00 [accepted] PHST- 2021/09/05 06:00 [pubmed] PHST- 2022/01/07 06:00 [medline] PHST- 2021/09/04 06:10 [entrez] AID - 10.1007/s00535-021-01820-0 [pii] AID - 10.1007/s00535-021-01820-0 [doi] PST - ppublish SO - J Gastroenterol. 2021 Nov;56(11):999-1007. doi: 10.1007/s00535-021-01820-0. Epub 2021 Sep 4.