1. IRF-1 mediates the suppressive effects of mTOR inhibition on arterial endothelium
- Author
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Peng, Kai, Fan, Xing, Li, Qiannan, Wang, Yiying, Chen, Xiaolin, Xiao, Pingxi, Passerini, Anthony G, Simon, Scott I, and Sun, ChongXiu
- Subjects
Biomedical and Clinical Sciences ,Medical Physiology ,Cardiovascular Medicine and Haematology ,Animals ,Apoptosis ,Cell Cycle Checkpoints ,Cell Proliferation ,Cells ,Cultured ,Endothelial Cells ,Endothelium ,Vascular ,Femoral Artery ,Humans ,Interferon Regulatory Factor-1 ,Mice ,Mice ,Knockout ,Naphthyridines ,Signal Transduction ,Sirolimus ,TOR Serine-Threonine Kinases ,Transfection ,Endothelium ,Cell proliferation ,Transcription factor ,Drug-eluting stents ,mTOR ,Cardiorespiratory Medicine and Haematology ,Cardiovascular System & Hematology ,Biochemistry and cell biology ,Cardiovascular medicine and haematology ,Medical physiology - Abstract
AimsMammalian target of rapamycin (mTOR) inhibitors used in drug-eluting stents (DES) to control restenosis have been found to delay endothelialization and increase incidence of late-stent thrombosis through mechanisms not completely understood. We revealed that mTOR inhibition (mTORi) upregulated the expression of cell growth suppressor IRF-1 in primary human arterial endothelial cells (HAEC). This study aimed to examine how mTOR-regulated IRF-1 expression contributes to the suppressive effect of mTORi on arterial endothelial proliferation.Methods and resultsWestern blotting, quantitative PCR, and a dual-luciferase reporter assay indicated that mTOR inhibitors rapamycin and torin 1 upregulated IRF-1 expression and increased its transcriptional activity. IRF-1 in turn contributed to the suppressive effect of mTORi by mediating HAEC apoptosis and cell cycle arrest in part through upregulation of caspase 1 and downregulation of cyclin D3, as revealed by CCK-8 assay, Annexin V binding assay, measurement of activated caspase 3, BrdU incorporation assay, and matrigel tube formation assay. In a mouse model of femoral artery wire injury, administration of rapamycin inhibited EC recovery, an effect alleviated by EC deficiency of IRF-1. Chromatin immunoprecipitation assay with HAEC and rescue expression of wild type or dominant-negative IRF-1 in EC isolated from Irf1-/- mice confirmed transcriptional regulation of IRF-1 on the expression of CASP1 and CCND3. Furthermore, mTORi activated multiple PKC members, among which PKCζ was responsible for the growth-inhibitory effect on HAEC. Activated PKCζ increased IRF1 transcription through JAK/STAT-1 and NF-κB signaling. Finally, overexpression of wild type or mutant raptor incapable of binding mTOR indicated that mTOR-free raptor contributed to PKCζ activation in mTOR-inhibited HAEC.ConclusionsThe study reveals an IRF-1-mediated mechanism that contributes to the suppressive effects of mTORi on HAEC proliferation. Further study may facilitate the development of effective strategies to reduce the side effects of DES used in coronary interventions.
- Published
- 2020