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Fibroblast-like synoviocyte migration is enhanced by IL-17-mediated overexpression of l-type amino acid transporter 1 (LAT1) via the mTOR/4E-BP1 pathway

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Abstract

In rheumatoid arthritis (RA), activated synovial fibroblasts have the ability to invade joint cartilage, actively contributing to joint destruction in RA. The mechanisms underlying this cell migration and invasion remain unclear. Our previous results and data from the GEO profile indicate that the l-type amino acid transporter gene, LAT1, is overexpressed in the synovium of RA. To identify its potential role in RA, fibroblast-like synoviocytes (FLS) from patients with RA were used to determine the effects of suppressing the LAT1 genes using RNA interference and the LAT inhibitor, BCH. We found that BCH exposure reduced the phosphorylation of mTOR and its downstream target 4EBP1, radiolabeled leucine uptake, and migration of RA FLS. LAT1 silencing by siRNA presented effects similar to BCH inhibition. Treatment of cells with IL-17 stimulated the expression of LAT1. In contrast, applying an inhibitor of mTOR pathway, temsirolimus, or silencing eIF4E neutralized the stimulation of IL-17 on LAT1. BCH and siLAT1 also resulted in lower IL-17-stimulated leucine uptake and cell migration. These results suggest that the migration of RA FLS is aggravated by IL-17-mediated overexpression of LAT1 via mTOR/4E-BP1 pathway. In conclusion, further investigation is warranted into LAT1 as a potential target for drug therapies aimed at attenuating migration of transformed-appearing fibroblasts and subsequently preventing further erosion of bone and cartilage.

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Acknowledgements

This study was supported by the National foundation of Natural Science (81671624), the Natural Science Foundation of Shandong Province (ZR2015YL029), and Shandong Health Science and Technology Research Program (2014WS0054), Key Project of Shandong Province (2017GSF218082), and Innovation Project of Shandong Academy of Medical Sciences.

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Authors and Affiliations

  1. Shandong Medicinal Biotechnology Centre, 18877 Jingshi Road, Jinan, Shandong, People’s Republic of China

    Zhao Yu, Wang Lin, Zhang Rui & Pan Jihong

  2. Key Lab for Biotechnology Drugs of Ministry of Health, 18877 Jingshi Road, Jinan, Shandong, People’s Republic of China

    Zhao Yu, Wang Lin, Zhang Rui & Pan Jihong

  3. Key Lab of Rare and Uncommon Disease, Shandong Province, 18877 Jingshi Road, Jinan, Shandong, People’s Republic of China

    Zhao Yu, Wang Lin, Zhang Rui & Pan Jihong

Authors
  1. Zhao Yu
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  2. Wang Lin
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  3. Zhang Rui
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  4. Pan Jihong
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Contributions

ZY performed the experiments; Dr. WL analyzed the data; ZR supplemented the experiments during revision; Dr. PJ designed the experiments and wrote the manuscript.

Corresponding author

Correspondence to Pan Jihong.

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Participants provided written informed consent to participate in the study and to allow their biological samples to be genetically analyzed.

Ethical standards

The Ethical Committee of Shandong Academy of Medicinal Sciences approved this study.

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The authors declare that they have no conflict of interest.

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Handling Editor: E. I. Closs.

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726_2017_2520_MOESM1_ESM.jpg

Supplementary material 1 (JPEG 50 kb) Supplementary Fig. 1 a IL-17 stimulated the expression of LAT1 in a dose-dependent manner for 12 h and the expression reached a maximum level with 50 ng/mL (***p < 0.001). b IL-17 stimulated the expression of LAT1 in time-dependent manner at a concentration of 50 ng/mL and the expression reached a maximum level in 12 h (***p < 0.001). All data are presented as the mean ± SEM of three independent experiments performed in triplicate

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Yu, Z., Lin, W., Rui, Z. et al. Fibroblast-like synoviocyte migration is enhanced by IL-17-mediated overexpression of l-type amino acid transporter 1 (LAT1) via the mTOR/4E-BP1 pathway. Amino Acids 50, 331–340 (2018). https://doi.org/10.1007/s00726-017-2520-4

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  • DOI: https://doi.org/10.1007/s00726-017-2520-4

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