Specificities of cellular uptake of primary cilium-derived extracellular vesicles for intercellular communication

细胞摄取原代纤毛来源的细胞外囊泡用于细胞间通讯的特异性

• 批准号: 21K15088
• 负责人: IJAZ FARYAL
• 金额: $ 3万
• 依托单位: Hiroshima University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Early-Career Scientists
• 财政年份: 2021
• 资助国家: 日本
• 起止时间: 2021-04-01 至 2024-03-31
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-21K15088/
• 关键词: Primary Cilium; Ciliopathies; primary cilium; Extracellular vesicles; Extracellular Vesicles

Cell-intrinsic mechanisms of ciliopathies have been well studied. In contrast, cell-extrinsic factors that underlie ciliopathies have barely been explored.We investigated the effects of extracellular bioactive factors derived from wildtype NIH/3T3 cells culture supernatant on the migration/ proliferation of primary cilium-deficient NIH/3T3-Kif3a-KO cells after wounding. The factors derived from wildtype NIH/3T3 cells culture supernatant increased the rate of cell migration/proliferation in target cells as compared to the factors derived from primary cilium-deficient cells culture supernatant from NIH/3T3-Kif3a-KO cells and NIH/3T3-Dync2h1-KO cells. Furthermore, by omics analysis of the supernatants, we found a molecule that showed a decline in the culture supernatant of primary cilium-deficient cells. These findings suggest that fibroblasts regulate cellular migration/proliferation process in primary cilium-deficient target cells via cell-extrinsic regulatory mechanisms in a primary cilium-dependent manner.

纤毛病的细胞内在机制已得到充分研究。相比之下，导致纤毛病的细胞外在因素几乎没有被探索过。我们研究了野生型NIH/3T3细胞培养上清液中衍生的细胞外生物活性因子对原代纤毛缺陷NIH/3T3-Kif3a-KO细胞受伤后迁移/增殖的影响。与源自NIH/3T3-Kif3a-KO细胞和NIH/3T3-Dync2h1-KO细胞的原代纤毛缺陷细胞培养上清液的因子相比，源自野生型NIH/3T3细胞培养上清液的因子增加了靶细胞中细胞迁移/增殖的速率。此外，通过对上清液的组学分析，我们发现了一种分子，其在原代纤毛缺陷细胞的培养上清液中表现出下降。这些发现表明，成纤维细胞通过细胞外在调节机制以原代纤毛依赖性方式调节原代纤毛缺陷的靶细胞中的细胞迁移/增殖过程。

项目成果

Double-strand break Site-Targeted PCR coupled with high concentration TBE-PAGE enables to detect NHEJ-mediated indel mutations at a 1-bp resolution

双链断裂定点 PCR 与高浓度 TBE-PAGE 相结合，能够以 1 bp 分辨率检测 NHEJ 介导的插入缺失突变

• 发表时间: 2022
• 作者: Faryal Ijaz;Ryota Nakazato;Mitsutoshi Setou;Koji Ikegami
• 通讯作者: Koji Ikegami

Time-lapse imaging of primary cilium behavior with physiological expression of fluorescent ciliary proteins

初级纤毛行为的延时成像与荧光纤毛蛋白的生理表达

• DOI: 10.1016/bs.mcb.2022.10.003
• 发表时间: 2023
• 期刊: Methods in Cell Biology
• 作者: Nakazato Ryota;Otani Hiroshi;Ijaz Faryal;Ikegami Koji
• 通讯作者: Ikegami Koji