論文 - 詳細
| RRC ID | 54868 |
|---|---|
| 著者 | Iwasawa C, Tamura R, Sugiura Y, Suzuki S, Kuzumaki N, Narita M, Suematsu M, Nakamura M, Yoshida K, Toda M, Okano H, Miyoshi H. |
| タイトル | Increased Cytotoxicity of Herpes Simplex Virus Thymidine Kinase Expression in Human Induced Pluripotent Stem Cells. |
| ジャーナル | Int J Mol Sci |
| Abstract |
Human induced pluripotent stem cells (iPSCs) hold enormous promise for regenerative medicine. The major safety concern is the tumorigenicity of transplanted cells derived from iPSCs. A potential solution would be to introduce a suicide gene into iPSCs as a safety switch. The herpes simplex virus type 1 thymidine kinase (HSV-TK) gene, in combination with ganciclovir, is the most widely used enzyme/prodrug suicide system from basic research to clinical applications. In the present study, we attempted to establish human iPSCs that stably expressed HSV-TK with either lentiviral vectors or CRISPR/Cas9-mediated genome editing. However, this task was difficult to achieve, because high-level and/or constitutive expression of HSV-TK resulted in the induction of cell death or silencing of HSV-TK expression. A nucleotide metabolism analysis suggested that excessive accumulation of thymidine triphosphate, caused by HSV-TK expression, resulted in an imbalance in the dNTP pools. This unbalanced state led to DNA synthesis inhibition and cell death in a process similar to a "thymidine block", but more severe. We also demonstrated that the Tet-inducible system was a feasible solution for overcoming the cytotoxicity of HSV-TK expression. Our results provided a warning against using the HSV-TK gene in human iPSCs, particularly in clinical applications. |
| 巻・号 | 20(4) |
| 公開日 | 2019-2-14 |
| DOI | 10.3390/ijms20040810 |
| PII | ijms20040810 |
| PMID | 30769780 |
| PMC | PMC6413063 |
| MeSH | Apoptosis / genetics CRISPR-Cas Systems / genetics Clustered Regularly Interspaced Short Palindromic Repeats Ganciclovir / pharmacology Gene Editing Gene Expression Regulation, Enzymologic / genetics Gene Expression Regulation, Viral / genetics Genes, Transgenic, Suicide / genetics Genetic Therapy* Genetic Vectors / therapeutic use Humans Induced Pluripotent Stem Cells / enzymology* Induced Pluripotent Stem Cells / transplantation Lentivirus / genetics Nucleotides / biosynthesis Nucleotides / genetics Simplexvirus / enzymology* Simplexvirus / genetics Thymidine Kinase / genetics* |
| IF | 4.183 |
| 引用数 | 6 |
| オルトメトリクス指標 |
オルトメトリクス指標項目
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| 各媒体での言及数の合計 | 0 |
| 過去6か月間でのオルトメトリクス指標の変動値 | 0.0 |
| リソース情報 | |
| ヒト・動物細胞 | NB1RGB(RCB0222) |