Loss of Nudt15 thiopurine detoxification increases direct DNA damage in hematopoietic stem cells
Abstract Thiopurines, such as 6-mercaptopurine (6-MP), are widely used as cytotoxic agents and immunosuppressants for leukemia and autoimmune or inflammatory diseases. A nonsynonymous single nucleotide polymorphism (p.Arg139Cys; R139C) of the nucleoside diphosphate-linked moiety X-type motif 15 (NUD...
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| Format: | Article |
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Nature Portfolio
2023-07-01
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| Series: | Scientific Reports |
| Online Access: | https://doi.org/10.1038/s41598-023-38952-7 |
| _version_ | 1797769560067145728 |
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| author | Noriaki Yamashita Masahiro Kawahara Takayuki Imai Goichi Tatsumi Ai Asai-Nishishita Akira Andoh |
| author_facet | Noriaki Yamashita Masahiro Kawahara Takayuki Imai Goichi Tatsumi Ai Asai-Nishishita Akira Andoh |
| author_sort | Noriaki Yamashita |
| collection | DOAJ |
| description | Abstract Thiopurines, such as 6-mercaptopurine (6-MP), are widely used as cytotoxic agents and immunosuppressants for leukemia and autoimmune or inflammatory diseases. A nonsynonymous single nucleotide polymorphism (p.Arg139Cys; R139C) of the nucleoside diphosphate-linked moiety X-type motif 15 (NUDT15) gene causes the loss of thiopurine detoxification, inducing myelosuppression. To understand such hematotoxicity, we investigate the effects of NUDT15 R139C on hematopoietic stem cells (HSCs) upon thiopurine administration. Using previously established Nudt15 R138C knock-in mice, which mimic myelosuppression in NUDT15 R139C homozygous or heterozygous patients following thiopurine administration, we investigated the numerical changes of HSCs and hematopoietic progenitor cells following 6-MP administration using in vivo flowcytometry and ex vivo HSC expansion. Genes differentially expressed between Nudt15 +/+ HSCs and Nudt15 R138C/R138C HSCs were identified using RNA-sequencing before the emergence of 6-MP-induced HSC-damage. Gene Ontology (GO) and Transcriptional Regulatory Relationships Unraveled by Sentence-based Text Mining (TRRUST) analyses were performed to elucidate the molecular effects of 6-MP on HSCs. In Nudt15 R138C/R138C mice, 6-MP induced exhaustion of HSCs faster than that of multipotent progenitors and as fast as that of myeloid-committed progenitors. Ex vivo-expanded Nudt15 R138C/R138C HSCs were dose- and time-dependently damaged by 6-MP. GO analysis identified the DNA damage response and cell cycle process as the most strongly influenced processes in Nudt15 R138C/R138C HSCs. TRRUST analysis revealed that the Trp53-regulated transcriptional regulatory network is influenced prior to HSC exhaustion in Nudt15 R138C/R138C HSCs. The loss of NUDT15 thiopurine detoxification enhances thiopurine-mediated DNA damage via the Trp53 networks in HSCs. Therefore, caution is required in long-term thiopurine use in patients with NUDT15 R139C in view of its adverse effects on HSCs in the form of DNA damage. |
| first_indexed | 2024-03-12T21:09:51Z |
| format | Article |
| id | doaj.art-3c96629815534cc8a1d0fa30ceebdd8c |
| institution | Directory Open Access Journal |
| issn | 2045-2322 |
| language | English |
| last_indexed | 2024-03-12T21:09:51Z |
| publishDate | 2023-07-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Scientific Reports |
| spelling | doaj.art-3c96629815534cc8a1d0fa30ceebdd8c2023-07-30T11:12:32ZengNature PortfolioScientific Reports2045-23222023-07-011311910.1038/s41598-023-38952-7Loss of Nudt15 thiopurine detoxification increases direct DNA damage in hematopoietic stem cellsNoriaki Yamashita0Masahiro Kawahara1Takayuki Imai2Goichi Tatsumi3Ai Asai-Nishishita4Akira Andoh5Division of Gastroenterology and Hematology, Department of Medicine, Shiga University of Medical ScienceDivision of Gastroenterology and Hematology, Department of Medicine, Shiga University of Medical ScienceDivision of Gastroenterology and Hematology, Department of Medicine, Shiga University of Medical ScienceDivision of Gastroenterology and Hematology, Department of Medicine, Shiga University of Medical ScienceDivision of Gastroenterology and Hematology, Department of Medicine, Shiga University of Medical ScienceDivision of Gastroenterology and Hematology, Department of Medicine, Shiga University of Medical ScienceAbstract Thiopurines, such as 6-mercaptopurine (6-MP), are widely used as cytotoxic agents and immunosuppressants for leukemia and autoimmune or inflammatory diseases. A nonsynonymous single nucleotide polymorphism (p.Arg139Cys; R139C) of the nucleoside diphosphate-linked moiety X-type motif 15 (NUDT15) gene causes the loss of thiopurine detoxification, inducing myelosuppression. To understand such hematotoxicity, we investigate the effects of NUDT15 R139C on hematopoietic stem cells (HSCs) upon thiopurine administration. Using previously established Nudt15 R138C knock-in mice, which mimic myelosuppression in NUDT15 R139C homozygous or heterozygous patients following thiopurine administration, we investigated the numerical changes of HSCs and hematopoietic progenitor cells following 6-MP administration using in vivo flowcytometry and ex vivo HSC expansion. Genes differentially expressed between Nudt15 +/+ HSCs and Nudt15 R138C/R138C HSCs were identified using RNA-sequencing before the emergence of 6-MP-induced HSC-damage. Gene Ontology (GO) and Transcriptional Regulatory Relationships Unraveled by Sentence-based Text Mining (TRRUST) analyses were performed to elucidate the molecular effects of 6-MP on HSCs. In Nudt15 R138C/R138C mice, 6-MP induced exhaustion of HSCs faster than that of multipotent progenitors and as fast as that of myeloid-committed progenitors. Ex vivo-expanded Nudt15 R138C/R138C HSCs were dose- and time-dependently damaged by 6-MP. GO analysis identified the DNA damage response and cell cycle process as the most strongly influenced processes in Nudt15 R138C/R138C HSCs. TRRUST analysis revealed that the Trp53-regulated transcriptional regulatory network is influenced prior to HSC exhaustion in Nudt15 R138C/R138C HSCs. The loss of NUDT15 thiopurine detoxification enhances thiopurine-mediated DNA damage via the Trp53 networks in HSCs. Therefore, caution is required in long-term thiopurine use in patients with NUDT15 R139C in view of its adverse effects on HSCs in the form of DNA damage.https://doi.org/10.1038/s41598-023-38952-7 |
| spellingShingle | Noriaki Yamashita Masahiro Kawahara Takayuki Imai Goichi Tatsumi Ai Asai-Nishishita Akira Andoh Loss of Nudt15 thiopurine detoxification increases direct DNA damage in hematopoietic stem cells Scientific Reports |
| title | Loss of Nudt15 thiopurine detoxification increases direct DNA damage in hematopoietic stem cells |
| title_full | Loss of Nudt15 thiopurine detoxification increases direct DNA damage in hematopoietic stem cells |
| title_fullStr | Loss of Nudt15 thiopurine detoxification increases direct DNA damage in hematopoietic stem cells |
| title_full_unstemmed | Loss of Nudt15 thiopurine detoxification increases direct DNA damage in hematopoietic stem cells |
| title_short | Loss of Nudt15 thiopurine detoxification increases direct DNA damage in hematopoietic stem cells |
| title_sort | loss of nudt15 thiopurine detoxification increases direct dna damage in hematopoietic stem cells |
| url | https://doi.org/10.1038/s41598-023-38952-7 |
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