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Microglia-Derived Spp1 Promotes Pathological Retinal Neovascularization via Activating Endothelial Kit/Akt/mTOR Signaling

Pathological retinal neovascularization (RNV) is the main character of ischemic ocular diseases, which causes severe visual impairments. Though retinal microglia are well acknowledged to play important roles in both physiological and pathological angiogenesis, the molecular mechanisms by which micro...

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Autores principales: Bai, Qian, Wang, Xin, Yan, Hongxiang, Wen, Lishi, Zhou, Ziyi, Ye, Yating, Jing, Yutong, Niu, Yali, Wang, Liang, Zhang, Zifeng, Su, Jingbo, Chang, Tianfang, Dou, Guorui, Wang, Yusheng, Sun, Jiaxing
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9866717/
https://www.ncbi.nlm.nih.gov/pubmed/36675807
http://dx.doi.org/10.3390/jpm13010146
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author Bai, Qian
Wang, Xin
Yan, Hongxiang
Wen, Lishi
Zhou, Ziyi
Ye, Yating
Jing, Yutong
Niu, Yali
Wang, Liang
Zhang, Zifeng
Su, Jingbo
Chang, Tianfang
Dou, Guorui
Wang, Yusheng
Sun, Jiaxing
author_facet Bai, Qian
Wang, Xin
Yan, Hongxiang
Wen, Lishi
Zhou, Ziyi
Ye, Yating
Jing, Yutong
Niu, Yali
Wang, Liang
Zhang, Zifeng
Su, Jingbo
Chang, Tianfang
Dou, Guorui
Wang, Yusheng
Sun, Jiaxing
author_sort Bai, Qian
collection PubMed
description Pathological retinal neovascularization (RNV) is the main character of ischemic ocular diseases, which causes severe visual impairments. Though retinal microglia are well acknowledged to play important roles in both physiological and pathological angiogenesis, the molecular mechanisms by which microglia communicates with endothelial cells (EC) remain unknown. In this study, using single-cell RNA sequencing, we revealed that the pro-inflammatory secreted protein Spp1 was the most upregulated gene in microglia in the mouse model of oxygen-induced retinopathy (OIR). Bioinformatic analysis showed that the expression of Spp1 in microglia was respectively regulated via nuclear factor-kappa B (NF-κB) and hypoxia-inducible factor 1α (HIF-1α) pathways, which was further confirmed through in vitro assays using BV2 microglia cell line. To mimic microglia-EC communication, the bEnd.3 endothelial cell line was cultured with conditional medium (CM) from BV2. We found that adding recombinant Spp1 to bEnd.3 as well as treating with hypoxic BV2 CM significantly enhanced EC proliferation and migration, while Spp1 neutralizing blocked those CM-induced effects. Moreover, RNA sequencing of BV2 CM-treated bEnd.3 revealed a significant downregulation of Kit, one of the type III tyrosine kinase receptors that plays a critical role in cell growth and activation. We further revealed that Spp1 increased phosphorylation and expression level of Akt/mTOR signaling cascade, which might account for its pro-angiogenic effects. Finally, we showed that intravitreal injection of Spp1 neutralizing antibody attenuated pathological RNV and improved visual function. Taken together, our work suggests that Spp1 mediates microglia-EC communication in RNV via activating endothelial Kit/Akt/mTOR signaling and is a potential target to treat ischemic ocular diseases.
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spelling pubmed-98667172023-01-22 Microglia-Derived Spp1 Promotes Pathological Retinal Neovascularization via Activating Endothelial Kit/Akt/mTOR Signaling Bai, Qian Wang, Xin Yan, Hongxiang Wen, Lishi Zhou, Ziyi Ye, Yating Jing, Yutong Niu, Yali Wang, Liang Zhang, Zifeng Su, Jingbo Chang, Tianfang Dou, Guorui Wang, Yusheng Sun, Jiaxing J Pers Med Article Pathological retinal neovascularization (RNV) is the main character of ischemic ocular diseases, which causes severe visual impairments. Though retinal microglia are well acknowledged to play important roles in both physiological and pathological angiogenesis, the molecular mechanisms by which microglia communicates with endothelial cells (EC) remain unknown. In this study, using single-cell RNA sequencing, we revealed that the pro-inflammatory secreted protein Spp1 was the most upregulated gene in microglia in the mouse model of oxygen-induced retinopathy (OIR). Bioinformatic analysis showed that the expression of Spp1 in microglia was respectively regulated via nuclear factor-kappa B (NF-κB) and hypoxia-inducible factor 1α (HIF-1α) pathways, which was further confirmed through in vitro assays using BV2 microglia cell line. To mimic microglia-EC communication, the bEnd.3 endothelial cell line was cultured with conditional medium (CM) from BV2. We found that adding recombinant Spp1 to bEnd.3 as well as treating with hypoxic BV2 CM significantly enhanced EC proliferation and migration, while Spp1 neutralizing blocked those CM-induced effects. Moreover, RNA sequencing of BV2 CM-treated bEnd.3 revealed a significant downregulation of Kit, one of the type III tyrosine kinase receptors that plays a critical role in cell growth and activation. We further revealed that Spp1 increased phosphorylation and expression level of Akt/mTOR signaling cascade, which might account for its pro-angiogenic effects. Finally, we showed that intravitreal injection of Spp1 neutralizing antibody attenuated pathological RNV and improved visual function. Taken together, our work suggests that Spp1 mediates microglia-EC communication in RNV via activating endothelial Kit/Akt/mTOR signaling and is a potential target to treat ischemic ocular diseases. MDPI 2023-01-11 /pmc/articles/PMC9866717/ /pubmed/36675807 http://dx.doi.org/10.3390/jpm13010146 Text en © 2023 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Bai, Qian
Wang, Xin
Yan, Hongxiang
Wen, Lishi
Zhou, Ziyi
Ye, Yating
Jing, Yutong
Niu, Yali
Wang, Liang
Zhang, Zifeng
Su, Jingbo
Chang, Tianfang
Dou, Guorui
Wang, Yusheng
Sun, Jiaxing
Microglia-Derived Spp1 Promotes Pathological Retinal Neovascularization via Activating Endothelial Kit/Akt/mTOR Signaling
title Microglia-Derived Spp1 Promotes Pathological Retinal Neovascularization via Activating Endothelial Kit/Akt/mTOR Signaling
title_full Microglia-Derived Spp1 Promotes Pathological Retinal Neovascularization via Activating Endothelial Kit/Akt/mTOR Signaling
title_fullStr Microglia-Derived Spp1 Promotes Pathological Retinal Neovascularization via Activating Endothelial Kit/Akt/mTOR Signaling
title_full_unstemmed Microglia-Derived Spp1 Promotes Pathological Retinal Neovascularization via Activating Endothelial Kit/Akt/mTOR Signaling
title_short Microglia-Derived Spp1 Promotes Pathological Retinal Neovascularization via Activating Endothelial Kit/Akt/mTOR Signaling
title_sort microglia-derived spp1 promotes pathological retinal neovascularization via activating endothelial kit/akt/mtor signaling
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9866717/
https://www.ncbi.nlm.nih.gov/pubmed/36675807
http://dx.doi.org/10.3390/jpm13010146
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