| 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
|
| リソース情報 |
| ヒト・動物細胞 |
NB1RGB(RCB0222) |
