PMID- 31200728 OWN - NLM STAT- MEDLINE DCOM- 20200615 LR - 20200615 IS - 1478-811X (Electronic) IS - 1478-811X (Linking) VI - 17 IP - 1 DP - 2019 Jun 14 TI - mTORC1 and mTORC2 are differentially engaged in the development of laser-induced CNV. PG - 64 LID - 10.1186/s12964-019-0380-0 [doi] LID - 64 AB - BACKGROUND: The mechanistic target of rapamycin (mTOR) pathway is a potential target to inhibit pathologic processes in choroidal neovascularization. However, the exact role of mTOR signaling in the development of CNV remains obscure. In this study, we assessed the role of mTORC1 and mTORC2 as well as the effect of rapamycin (sirolimus) on choroidal neovascularization (CNV) in a laser-induced mouse model. METHODS: In experiment A, we observed the natural course of CNV development and the dynamics of mTOR-related proteins during the 12 days after the laser injury. The expression of mTOR-related proteins was evaluated using Western blot (WB). Cryosections of CNV-induced mice were immunostained for the visualization of the vascular and extravascular components of the CNV. Experiment B was performed to confirm the critical period of mTOR signaling in the development of laser-induced CNV, we administered rapamycin before and/or during the active period of mTOR complexes. WB and immunofluorescence staining was performed to evaluate the mode of action and the effect of mTOR inhibition on CNV development. RESULTS: In experiment A, we detected high levels of p-mTOR S2448 and p-mTOR S2481 from the 5th to 12th day of laser injury. Immunofluorescence imaging of cryosections of mice sacrificed on day 7 revealed greater co-immunoreactivity of p-mTOR S2448 positive cells with CD11b and F4/80, while p-mTOR S2481 positive cells showed colocalization with CD31, alpha-SMA, and cytokeratin. In experiment B, rapamycin injection during the active period of mTOR signaling demonstrated near-complete inhibition of CNV lesion as well as significant induction of autophagy. CONCLUSION: Our study suggests the mTOR as a critical player during CNV development in laser-induced mouse model through differentially acting with the mTORC1 and mTORC2. mTORC1 activity was high predominantly in inflammatory cells in CNV lesion, while mTORC2 activity was higher in vascular components and the RPE. FAU - Yang, Jin Young AU - Yang JY AD - Department of Interdisciplinary Program in Biomedical Science, Soonchunhyang Graduate School, Bucheon Hospital, Bucheon, South Korea. AD - Laboratory for Translational Research on Retinal and Macular Degeneration, Soonchunhyang University Hospital Bucheon, Bucheon, South Korea. FAU - Madrakhimov, Sanjar Batirovich AU - Madrakhimov SB AD - Department of Interdisciplinary Program in Biomedical Science, Soonchunhyang Graduate School, Bucheon Hospital, Bucheon, South Korea. AD - Laboratory for Translational Research on Retinal and Macular Degeneration, Soonchunhyang University Hospital Bucheon, Bucheon, South Korea. FAU - Ahn, Dong Hyuck AU - Ahn DH AD - Laboratory for Translational Research on Retinal and Macular Degeneration, Soonchunhyang University Hospital Bucheon, Bucheon, South Korea. FAU - Chang, Hun Soo AU - Chang HS AD - Department of Medical Bioscience, Graduated School, Soonchunhyang University, Bucheon, South Korea. FAU - Jung, Sang Joon AU - Jung SJ AD - Department of Ophthalmology, College of Medicine, Soonchunhyang University, Cheonan, Choongchungnam-do, South Korea. FAU - Nah, Seung Kwan AU - Nah SK AD - Department of Ophthalmology, College of Medicine, Soonchunhyang University, Cheonan, Choongchungnam-do, South Korea. FAU - Park, Ha Yan AU - Park HY AD - Laboratory for Translational Research on Retinal and Macular Degeneration, Soonchunhyang University Hospital Bucheon, Bucheon, South Korea. FAU - Park, Tae Kwann AU - Park TK AUID- ORCID: 0000-0001-9689-4384 AD - Department of Interdisciplinary Program in Biomedical Science, Soonchunhyang Graduate School, Bucheon Hospital, Bucheon, South Korea. tkpark@schmc.ac.kr. AD - Laboratory for Translational Research on Retinal and Macular Degeneration, Soonchunhyang University Hospital Bucheon, Bucheon, South Korea. tkpark@schmc.ac.kr. AD - Department of Ophthalmology, Soonchunhyang University Hospital Bucheon, #170, Jomaru-ro, Wonmi-gu, Bucheon, 14584, South Korea. tkpark@schmc.ac.kr. AD - Department of Ophthalmology, College of Medicine, Soonchunhyang University, Cheonan, Choongchungnam-do, South Korea. tkpark@schmc.ac.kr. AD - Department of Ophthalmology, College of Medicine, Soonchunhyang University, Bucheon Hospital, Bucheon, South Korea. tkpark@schmc.ac.kr. LA - eng PT - Journal Article PT - Research Support, Non-U.S. Gov't DEP - 20190614 PL - England TA - Cell Commun Signal JT - Cell communication and signaling : CCS JID - 101170464 RN - EC 2.7.11.1 (Mechanistic Target of Rapamycin Complex 1) RN - EC 2.7.11.1 (Mechanistic Target of Rapamycin Complex 2) RN - W36ZG6FT64 (Sirolimus) SB - IM MH - Animals MH - Autophagy/drug effects/radiation effects MH - Choroidal Neovascularization/etiology/*metabolism/pathology MH - Lasers/*adverse effects MH - Male MH - Mechanistic Target of Rapamycin Complex 1/antagonists & inhibitors/*metabolism MH - Mechanistic Target of Rapamycin Complex 2/antagonists & inhibitors/*metabolism MH - Mice MH - Mice, Inbred C57BL MH - Signal Transduction/radiation effects MH - Sirolimus/pharmacology PMC - PMC6570852 OTO - NOTNLM OT - Age-related macular degeneration OT - Choroidal neovascularization OT - Sirolimus (rapamycin) OT - mTORC1 OT - mTORC2 COIS- The authors declare that they have no competing interests. EDAT- 2019/06/16 06:00 MHDA- 2020/06/17 06:00 PMCR- 2019/06/14 CRDT- 2019/06/16 06:00 PHST- 2019/01/28 00:00 [received] PHST- 2019/06/04 00:00 [accepted] PHST- 2019/06/16 06:00 [entrez] PHST- 2019/06/16 06:00 [pubmed] PHST- 2020/06/17 06:00 [medline] PHST- 2019/06/14 00:00 [pmc-release] AID - 10.1186/s12964-019-0380-0 [pii] AID - 380 [pii] AID - 10.1186/s12964-019-0380-0 [doi] PST - epublish SO - Cell Commun Signal. 2019 Jun 14;17(1):64. doi: 10.1186/s12964-019-0380-0.