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Soluble delta-like 1 homolog (DLK1) stimulates angiogenesis through Notch1/Akt/eNOS signaling in endothelial cells

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A Correction to this article was published on 10 May 2018

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Abstract

Aim

Delta-like 1 homolog (DLK1) is a non-canonical ligand of Notch signaling, which plays a pivotal role in vascular development and tumor angiogenesis. This study aimed to elucidate the function and mechanism of DLK1 in angiogenesis.

Methods and results

By using in situ hybridization and immunohistochemical studies, expression analysis revealed a unique vascular tropism of DLK1 in vasculature of neuroblastoma and vascular tumors. Thus, it was hypothesized that DLK1 may be cleaved and then bound to endothelial cells, thereby regulating the endothelial function. To test such hypothesis, soluble DLK1 encompassing DLK1 extracellular domain (DLK1-EC) was generated and validated by its inhibitory function in adipogenesis assay. Recombinant DLK1-EC exhibited the preferential binding capability toward endothelial cells and stimulated the microvessels sprouting in aorta rings. Above all, implantation of DLK1-EC dose-dependently elicited the cornea neovascularization in rats. By using various angiogenesis assays, it was delineated that DLK1-EC stimulated the angiogenesis by promoting the proliferation, motility and tube formation of endothelial cells. By immunoblot and luciferase analysis, it was elucidated that DLK1-EC enhanced the expression and activities of Notch1/Akt/eNOS/Hes-1 signaling in dose- and time-dependent manners. Pharmaceutical blockage of Notch signaling using γ-secretase inhibitor DAPT abrogated the DLK1-EC-induced endothelial migration and Hes-1-driven luciferase activities. Furthermore, Notch1 inactivation by neutralizing antibodies or RNA interference reversed the DLK1-EC-induced angiogenesis.

Conclusions

The present study unveils the pro-angiogenic function and mechanism of soluble DLK1 through activation of Notch1 signaling in endothelial cells.

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Change history

  • 10 May 2018

    In the original publication of the article, there is an error in one of the citations in the Discussion section.

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Acknowledgements

We thank Chang Gung Medical Foundation and Tissue Bank of Kaohsiung Chang Gung Memorial Hospital (CLRPG8B0031 and CLRPG8B0033) for technical support.

Funding

This work was supported by Grants from the Ministry of Science and Technology, Taiwan (MOST 102-2325-B-110-001 to MHT, MOST 98-2320-B-182-023-MY3 and MOST 101-2320-B-182-009 to CCH), Chang Gung Memorial Hospital (CMRPG890303 and CMRPG891091), NSYSU-KMU Joint Research Project (NSYSUKMU104-P039) and the Center for Stem Cell Research, Kaohsiung Medical University, Taiwan (KMU-TP105G01).

Author information

Author notes

  1. Chao-Cheng Huang and Hsiao-Mei Kuo have contributed equally to this work.

Authors and Affiliations

  1. Biobank and Tissue Bank and Department of Pathology, Kaohsiung Chang Gung Memorial Hospital, Kaohsiung, Taiwan

    Chao-Cheng Huang & Shih-Hsuan Cheng

  2. Graduate Institute of Clinical Medical Sciences, Chang Gung University College of Medicine, Kaohsiung, Taiwan

    Chao-Cheng Huang

  3. Center for Neuroscience, National Sun Yat-Sen University, Kaohsiung, Taiwan

    Hsiao-Mei Kuo & Ming-Hong Tai

  4. Department of Biological Sciences, National Sun Yat-Sen University, Kaohsiung, Taiwan

    Hsiao-Mei Kuo, Tzu-Ting Chang & Ming-Hong Tai

  5. Departments of Ophthalmology, Kaohsiung Chang Gung Memorial Hospital, Kaohsiung, Taiwan

    Pei-Chang Wu

  6. Institute of Chemistry, Academia Sinica, Taipei, Taiwan

    Tzu-Ting Chang

  7. Doctoral Degree Program in Marine Biotechnology, National Sun Yat-Sen University, Kaohsiung, Taiwan

    Yi-Chen Chang & Ming-Hong Tai

  8. Doctoral Degree Program in Marine Biotechnology, Academia Sinica, Taipei, Taiwan

    Yi-Chen Chang & Ming-Hong Tai

  9. Department of Chemistry, National Sun Yat-Sen University, Kaohsiung, Taiwan

    Mei-Lang Kung

  10. Center for Stem Cell Research, Department of Internal Medicine, Kaohsiung Medical University, Kaohsiung, Taiwan

    Deng-Chyang Wu & Ming-Hong Tai

  11. Division of Gastroenterology, Department of Internal Medicine, Kaohsiung Medical University, Kaohsiung, Taiwan

    Shih-Hsuan Cheng, Deng-Chyang Wu & Ming-Hong Tai

  12. Division of Pediatric Surgery, Department of Surgery, Kaohsiung Chang Gung Memorial Hospital, Kaohsiung, 833, Taiwan

    Jiin-Haur Chuang

  13. Institute of Biomedical Sciences, National Sun Yat-Sen University, 70 Lien-Hai Rd, Kaohsiung, 804, Taiwan

    Ming-Hong Tai

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  1. Chao-Cheng Huang
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Correspondence to Jiin-Haur Chuang or Ming-Hong Tai.

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Huang, CC., Kuo, HM., Wu, PC. et al. Soluble delta-like 1 homolog (DLK1) stimulates angiogenesis through Notch1/Akt/eNOS signaling in endothelial cells. Angiogenesis 21, 299–312 (2018). https://doi.org/10.1007/s10456-018-9596-7

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