Hypoxic preconditioning of human cardiosphere-derived cell sheets enhances cellular functions via activation of the PI3K/Akt/mTOR/HIF-1α pathway.
Hypoxic preconditioning of human cardiosphere-derived cell sheets enhances cellular functions via activation of the PI3K/Akt/mTOR/HIF-1α pathway.
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DOI：
10.20944/preprints201608.0124.v1
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发表时间：
2016-08
影响因子：
2.2
通讯作者：
Yuya Tanaka;Tohru Hosoyama;A. Mikamo;H. Kurazumi;Arata Nishimoto;Koji Ueno;B. Shirasawa;K. Hamano
中科院分区：
文献类型：
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作者：
Yuya Tanaka;Tohru Hosoyama;A. Mikamo;H. Kurazumi;Arata Nishimoto;Koji Ueno;B. Shirasawa;K. Hamano
细胞片层技术是一种很有前途的治疗策略，用于治疗缺血性疾病，如心肌梗死。我们最近开发了一种新的协议，称为“缺氧预处理”，能够增强细胞片的治疗效果。根据该方案，通过在缺氧培养条件下短暂孵育细胞片来增强细胞片的促血管生成和抗纤维化活性。然而，细胞片层缺氧预适应的确切分子机制尚不清楚。在本研究中，我们研究了信号转导细胞片，以确定那些响应缺氧预处理，使用心肌球衍生的细胞（CDC）片。我们最初测试片状结构是否适合缺氧预处理，通过比较它们与单个细胞。低渗预处理对片状细胞比单个细胞更有效。缺氧预处理诱导细胞表达缺氧诱导因子-1 α（HIF-1α）和哺乳动物雷帕霉素靶蛋白（mTOR），并诱导磷酸肌醇3-激酶（PI 3 K）/Akt通路。此外，低氧预处理增加了表皮生长因子受体（EGFR）和热休克蛋白60（HSP 60）在CDC片的磷酸化。我们的研究结果提供了新的见解，利用缺氧预处理的细胞片为基础的技术，用于治疗缺血性疾病。
Cell sheet technology is a promising therapeutic strategy for the treatment of ischemic diseases such as myocardial infarction. We recently developed a novel protocol, termed "hypoxic preconditioning," capable of augmenting the therapeutic efficacy of cell sheets. Following this protocol, the pro-angiogenic and anti-fibrotic activity of cell sheets were enhanced by brief incubation of cell sheets under hypoxic culture conditions. However, the precise molecular mechanism underlying the hypoxic preconditioning of cell sheets is unclear. In the present study, we examined signal transducers in cell sheets to identify those responsive to hypoxic preconditioning, using cardiosphere-derived cell (CDC) sheets. We initially tested whether sheet-like structures were suitable for hypoxic preconditioning by comparing them with individual cells. Hypoxic preconditioning was more effective in sheeted cells than in individual cells. Expression of hypoxia inducible factor-1α (HIF-1α) and mammalian target of rapamycin (mTOR) were induced upon hypoxic preconditioning of cell sheets, as was the phosphoinositide 3-kinase (PI3K)/Akt pathway. In addition, hypoxic preconditioning increased phosphorylation of epidermal growth factor receptor (EGFR) and heat shock protein 60 (HSP60) in CDC sheets. Our findings provide novel insights into the utility of hypoxic preconditioning in cell sheet-based technologies for the treatment of ischemic diseases.