Xin Wang1, Chang Chun Ling2, Liping Li1, Yinyin Qin1, Jialing Qi3, Xiaoyu Liu1, Bo You2, Yunwei Shi1, Jie Zhang3, Qiu Jiang4, Hui Xu1, Cheng Sun1, Yiwen You2, Renjie Chai5, Dong Liu6. 1. Co-innovation Center of Neuroregeneration, Jiangsu Key Laboratory of Neuroregeneration, Nantong University, Qixiu Road 19, Nantong 226001, China. 2. Affiliated Hospital of Nantong University, Nantong, China. 3. Medical College, Nantong University, Nantong, China. 4. Shanghai Institute of Cardiovascular Diseases, Zhongshan Hospital and Institutes of Biomedical Sciences, Fudan University, Shanghai, China. 5. Co-innovation Center of Neuroregeneration, Jiangsu Key Laboratory of Neuroregeneration, Nantong University, Qixiu Road 19, Nantong 226001, China Co-innovation Center of Neuroregeneration, Key Laboratory for Developmental Genes and Human Disease, Ministry of Education, Institute of Life Sciences, Southeast University, No. 2 Sipailou Road, Nanjing 210096, China liudongtom@gmail.com renjiec@seu.edu.cn. 6. Co-innovation Center of Neuroregeneration, Jiangsu Key Laboratory of Neuroregeneration, Nantong University, Qixiu Road 19, Nantong 226001, China liudongtom@gmail.com renjiec@seu.edu.cn.
Abstract
AIMS: MicroRNA-10 (miR-10) was originally shown to regulate angiogenesis by directly modulating the levels of membrane-bound fms-related tyrosine kinase 1 (mflt1) and its soluble splice isoform sflt1 post-transcriptionally in zebrafish. Given that flt1 knockdown incompletely rescues the angiogenic phenotypes in miR-10 morphants, flt1 is unlikely to be the only important target of miR-10 in endothelial cells (ECs). It will be interesting to investigate new mechanism responsible for angiogenic defect induced by miR-10 knockdown. METHODS AND RESULTS: Firstly, we demonstrated that miR-10a and miR-10b (miR-10a/10b) were highly enriched in embryonic zebrafish ECs using deep sequencing, Taqman polymerase chain reaction, and in situ hybridisation. Subsequently, we proved that loss of miR-10a/10b impaired blood vessel outgrowth through regulating tip cell behaviours. Mib1 was identified as a potential direct target of miR-10a/10b through in silicon analysis and in vitro luciferase sensor assay. In vivo reporter assay in zebrafish embryos confirmed the binding of miR-10 with 3'-UTR of zebrafish mib1. Furthermore, inhibition of mib1 and Notch signaling rescued the angiogenic defects in miR-10-deficient zebrafish embryos. In addition, we provided evidences that miR-10 regulates human ECs behaviour through targeting Mib1 as well. CONCLUSION: Taken together, these results indicate that miR-10 regulates the angiogenic behaviour in a Notch-dependent manner by directly targeting mib1. Published on behalf of the European Society of Cardiology. All rights reserved.
AIMS: MicroRNA-10 (miR-10) was originally shown to regulate angiogenesis by directly modulating the levels of membrane-bound fms-related tyrosine kinase 1 (mflt1) and its soluble splice isoform sflt1 post-transcriptionally in zebrafish. Given that flt1 knockdown incompletely rescues the angiogenic phenotypes in miR-10 morphants, flt1 is unlikely to be the only important target of miR-10 in endothelial cells (ECs). It will be interesting to investigate new mechanism responsible for angiogenic defect induced by miR-10 knockdown. METHODS AND RESULTS: Firstly, we demonstrated that miR-10a and miR-10b (miR-10a/10b) were highly enriched in embryonic zebrafish ECs using deep sequencing, Taqman polymerase chain reaction, and in situ hybridisation. Subsequently, we proved that loss of miR-10a/10b impaired blood vessel outgrowth through regulating tip cell behaviours. Mib1 was identified as a potential direct target of miR-10a/10b through in silicon analysis and in vitro luciferase sensor assay. In vivo reporter assay in zebrafish embryos confirmed the binding of miR-10 with 3'-UTR of zebrafishmib1. Furthermore, inhibition of mib1 and Notch signaling rescued the angiogenic defects in miR-10-deficient zebrafish embryos. In addition, we provided evidences that miR-10 regulates human ECs behaviour through targeting Mib1 as well. CONCLUSION: Taken together, these results indicate that miR-10 regulates the angiogenic behaviour in a Notch-dependent manner by directly targeting mib1. Published on behalf of the European Society of Cardiology. All rights reserved.