1. AAV expressing an mTOR‐inhibiting siRNA exhibits therapeutic potential in retinal vascular disorders by preserving endothelial integrity
- Author
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Heuiran Lee, Won-Il Seo, Steven Hyun Seung Lee, Jun-Sub Choi, Hee Jong Kim, Ha-Na Woo, Joo Yong Lee, Jin Kim, Keerang Park, Beom Jun Lee, and Seho Cha
- Subjects
Vascular Endothelial Growth Factor A ,Angiogenesis ,QH301-705.5 ,tight junction ,viruses ,proliferation ,mTORC1 ,medicine.disease_cause ,migration ,mTORC2 ,General Biochemistry, Genetics and Molecular Biology ,Neovascularization ,retinal vascular disorder ,chemistry.chemical_compound ,angiogenesis ,Human Umbilical Vein Endothelial Cells ,medicine ,Humans ,RNA, Small Interfering ,Biology (General) ,Mechanistic target of rapamycin ,Adeno-associated virus ,Research Articles ,PI3K/AKT/mTOR pathway ,biology ,TOR Serine-Threonine Kinases ,adeno‐associated virus ,Dependovirus ,endothelial cells ,small‐hairpin mTOR ,Vascular endothelial growth factor ,chemistry ,Cancer research ,biology.protein ,medicine.symptom ,Research Article - Abstract
Expanding on previous demonstrations of the therapeutic effects of adeno‐associated virus (AAV) carrying small‐hairpin RNA (shRNA) in downregulating the mechanistic target of rapamycin (mTOR) in in vivo retinal vascular disorders, vascular endothelial growth factor (VEGF)‐stimulated endothelial cells were treated with AAV2‐shmTOR to examine the role of mTOR inhibition in retinal angiogenesis. AAV2‐shmTOR exposure significantly reduced mTOR expression in human umbilical vein endothelial cells (HUVECs) and decreased downstream signaling cascades of mTOR complex 1 (mTORC1) and mTORC2 under VEGF treatment. Moreover, the angiogenic potential of VEGF was significantly inhibited by AAV2‐shmTOR, which preserved endothelial integrity by maintaining tight junctions between HUVECs. These data thus support previous in vivo studies and provide evidence that AAV2‐shmTOR induces therapeutic effects by inhibiting the neovascularization of endothelial cells., Inhibition of mTOR expression by AAV2‐shmTOR suppresses the proliferation and migration of endothelial cells stimulated by VEGF. mTOR is a major activator in endothelial cells under VEGF stimulation, and inhibition by AAV2‐shmTOR blocked the activation of downstream mTOR signaling molecules. Inhibition of the mTOR pathway consequently affected cell proliferation and the remodeling of tight junctions between endothelial cells.
- Published
- 2022