論文 - 詳細
| RRC ID | 44662 |
|---|---|
| 著者 | Saka K, Kawahara M, Teng J, Otsu M, Nakauchi H, Nagamune T. |
| タイトル | Top-down motif engineering of a cytokine receptor for directing ex vivo expansion of hematopoietic stem cells. |
| ジャーナル | J Biotechnol |
| Abstract |
The technique to expand hematopoietic stem cells (HSCs) ex vivo is eagerly anticipated to secure an enough amount of HSCs for clinical applications. Previously we developed a scFv-thrombopoietin receptor (c-Mpl) chimera, named S-Mpl, which can transduce a proliferation signal in HSCs in response to a cognate antigen. However, a remaining concern of the S-Mpl chimera may be the magnitude of the cellular expansion level driven by this molecule, which was significantly less than that mediated by endogenous wild-type c-Mpl. In this study, we engineered a tyrosine motif located in the intracellular domain of S-Mpl based on a top-down approach in order to change the signaling properties of the chimera. The truncated mutant (trunc.) and an amino-acid substitution mutant (Q to L) of S-Mpl were constructed to investigate the ability of these mutants to expand HSCs. The result showed that the truncated and Q to L mutants gave higher and considerably lower number of the cells than unmodified S-Mpl, respectively. The proliferation level through the truncated mutant was even higher than that of non-transduced HSCs with the stimulation of a native cytokine, thrombopoietin. Moreover, we analyzed the signaling properties of the S-Mpl mutants in detail using a pro-B cell line Ba/F3. The data indicated that the STAT3 and STAT5 activation levels through the truncated mutant increased, whereas activation of the Q to L mutant was inhibited by a negative regulator of intracellular signaling, SHP-1. This is the first demonstration that a non-natural artificial mutant of a cytokine receptor is effective for ex vivo expansion of hematopoietic cells compared with a native cytokine receptor. |
| 巻・号 | 168(4) |
| ページ | 659-65 |
| 公開日 | 2013-12-1 |
| DOI | 10.1016/j.jbiotec.2013.09.012 |
| PII | S0168-1656(13)00403-3 |
| PMID | 24070902 |
| MeSH | Amino Acid Motifs / genetics Gene Expression Regulation Genetic Engineering* Hematopoietic Stem Cells / cytology Hematopoietic Stem Cells / metabolism* Humans Mutation Receptors, Thrombopoietin / genetics* Receptors, Thrombopoietin / immunology Receptors, Thrombopoietin / metabolism Recombinant Fusion Proteins / genetics* Recombinant Fusion Proteins / immunology Recombinant Fusion Proteins / metabolism STAT3 Transcription Factor / biosynthesis STAT3 Transcription Factor / genetics STAT5 Transcription Factor / biosynthesis STAT5 Transcription Factor / genetics Signal Transduction Thrombopoietin / immunology Thrombopoietin / metabolism |
| IF | 3.503 |
| 引用数 | 2 |
| WOS 分野 | BIOTECHNOLOGY & APPLIED MICROBIOLOGY |
| 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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| 最多言及媒体 | Patent(IFI CLAIMS) |
| 各媒体での言及数の合計 | 6 |
| 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 | |