論文 - 詳細
| RRC ID | 72199 |
|---|---|
| 著者 | Tang SY, Zha S, Du Z, Zeng J, Zhu D, Luo Y, Wang S. |
| タイトル | Targeted integration of EpCAM-specific CAR in human induced pluripotent stem cells and their differentiation into NK cells. |
| ジャーナル | Stem Cell Res Ther |
| Abstract |
BACKGROUND:Redirection of natural killer (NK) cells with chimeric antigen receptors (CAR) is attractive in developing off-the-shelf CAR therapeutics for cancer treatment. However, the site-specific integration of a CAR gene into NK cells remains challenging. METHODS:In the present study, we genetically modified human induced pluripotent stem cells (iPSCs) with a zinc finger nuclease (ZFN) technology to introduce a cDNA encoding an anti-EpCAM CAR into the adeno-associated virus integration site 1, a "safe harbour" for transgene insertion into human genome, and next differentiated the modified iPSCs into CAR-expressing iNK cells. RESULTS:We detected the targeted integration in 4 out of 5 selected iPSC clones, 3 of which were biallelically modified. Southern blotting analysis revealed no random integration events. iNK cells were successfully derived from the modified iPSCs with a 47-day protocol, which were morphologically similar to peripheral blood NK cells, displayed NK phenotype (CD56+CD3-), and expressed NK receptors. The CAR expression of the iPSC-derived NK cells was confirmed with RT-PCR and flow cytometry analysis. In vitro cytotoxicity assay further confirmed their lytic activity against NK cell-resistant, EpCAM-positive cancer cells, but not to EpCAM-positive normal cells, demonstrating the retained tolerability of the CAR-iNK cells towards normal cells. CONCLUSION:Looking ahead, the modified iPSCs generated in the current study hold a great potential as a practically unlimited source to generate anti-EpCAM CAR iNK cells. |
| 巻・号 | 12(1) |
| ページ | 580 |
| 公開日 | 2021-11-21 |
| DOI | 10.1186/s13287-021-02648-4 |
| PII | 10.1186/s13287-021-02648-4 |
| PMID | 34802459 |
| PMC | PMC8607711 |
| MeSH | Cell Differentiation Epithelial Cell Adhesion Molecule / genetics Epithelial Cell Adhesion Molecule / metabolism Humans Induced Pluripotent Stem Cells* / metabolism Killer Cells, Natural Receptors, Chimeric Antigen* / genetics Receptors, Chimeric Antigen* / metabolism |
| IF | 5.116 |
| リソース情報 | |
| ヒト・動物細胞 | OP9/G-DLL1(RCB2925) |