generation of functional CX26 gap junction forming cell derived from mouse iPS cells and its application to hereditary deafness
| Project/Area Number |
17K16948
|
|---|
| Research Category |
Grant-in-Aid for Young Scientists (B)
|
| Allocation Type | Multi-year Fund |
|---|
| Research Field |
Otorhinolaryngology
|
|---|
| Research Institution | Juntendo University |
|---|
Principal Investigator |
Fukunaga Ichiro 順天堂大学, 医学(系)研究科(研究院), 助教 (20746581)
|
|---|
| Project Period (FY) |
2017-04-01 – 2020-03-31
|
| Project Status |
Completed (Fiscal Year 2019)
|
| Budget Amount *help |
¥4,030,000 (Direct Cost: ¥3,100,000、Indirect Cost: ¥930,000)
Fiscal Year 2018: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000)
Fiscal Year 2017: ¥2,340,000 (Direct Cost: ¥1,800,000、Indirect Cost: ¥540,000)
|
|---|
| Keywords | GJB2 / 遺伝性難聴 / ES/iPS細胞 / 分化誘導 / 鳥類胚 / 内耳細胞 / Connexin26 / iPS細胞 / 蝸牛支持細胞 / ギャップ結合プラーク |
|---|
| Outline of Final Research Achievements |
Mutation of the Gap Junction Beta 2 gene (GJB2) encoding connexin 26 (CX26) is the most frequent cause of hereditary deafness worldwide. In this study, we generated functional CX26 gap junction-forming cell (iCX26GJC) from mouse ESC/iPSC by using 3D and 2D culture. In 3D culture, iCX6GJC were observed in a part of the aggregate. Aggregates were subcultured in 2D culture, and proliferation of iCX26GJCs were observed. To investigate whether the iCx26GJC were functional, we performed scrape-loading assay and Ca2+ imaging. In the iCX26GJC culture, we observed that Lucifer yellow diffused beyond the wounded parental cells. Furthermore, spontaneous Ca2+ signaling activity was observed. Finally, we found that combination with growth factor and inhibitor has been shown to result in greater production of isolatable CX26-expressing region and higher Gjb2 mRNA levels, thereby increasing the yield of highly purified iCX26GJCs.
|
| Academic Significance and Societal Importance of the Research Achievements |
これまで、ES/iPS細胞から感覚細胞への分化誘導法は複数報告されているが、CX26ギャップ結合を構築する蝸牛支持細胞を作製した報告はなかった。CX26をコードするGJB2遺伝子は世界最大の遺伝性難聴の原因遺伝子であり、疾患の対象となる細胞は蝸牛支持細胞などの非感覚細胞である。本研究の成果は、GJB2変異難聴を含めた、非感覚細胞を対象とする難聴の病態の解明や、治療法の開発への応用が期待される。
|
Report
(4 results)
Research Products
(7 results)
