Inhibiting HIF-1 signaling alleviates HTRA1-induced RPE senescence in retinal degeneration.
Inhibiting HIF-1 signaling alleviates HTRA1-induced RPE senescence in retinal degeneration.
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DOI：
10.1186/s12964-023-01138-9
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发表时间：
2023-06-14
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影响因子：
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视网膜色素上皮细胞（retinal pigment epithelium，RPE）和感光细胞的变性是导致老年人不可逆视力损害的主要原因。视网膜色素上皮细胞的衰老是导致AMD的重要原因之一，并已成为AMD治疗的潜在靶点。HTRA 1是AMD最重要的易感基因之一，但在AMD发病机制中，HTRA 1与RPE衰老的相关性尚未见报道。采用Western blotting和免疫组织化学方法检测WT和过表达人HTRA 1的转基因小鼠（hHTRA 1-Tg小鼠）中HTRA 1的表达。RT-qPCR检测hHTRA 1-Tg小鼠和HTRA 1感染的ARPE-19细胞中SASP的表达。透射电镜、SA-β-gal法观察RPE细胞线粒体的变化及细胞的衰老。通过眼底照相、FFA、SD-OCT和ERG观察小鼠视网膜变性。分析用adv-HTRA 1相对于adv-NC处理的ARPE-19细胞的RNA-Seq数据集。用OCR和ECAR测定ARPE-19细胞线粒体呼吸和糖酵解能力。用EF 5缺氧检测试剂盒检测ARPE-19细胞的缺氧情况。KC 7 F2用于降低体外和体内HIF 1 α的表达。在我们的研究中，我们发现RPE衰老促进hHTRA 1-Tg小鼠。hHTRA 1-Tg小鼠在氧化应激诱导的视网膜变性的发展中变得更容易受到NaIO 3的影响。类似地，HTRA 1在ARPE-19细胞中的过表达加速了细胞衰老。我们的RNA-seq揭示了HTRA 1诱导的与衰老相关的差异表达基因与ARPE-19细胞中参与线粒体功能和缺氧反应的基因之间的重叠。HTRA 1在ARPE-19细胞中的过表达损害了线粒体功能并增强了糖酵解能力。重要的是，HTRA 1的上调显著激活了HIF-1信号通路，表现为促进主要位于细胞核的HIF 1 α的表达。HIF 1 α翻译抑制剂KC 7 F2显著阻止HTRA 1诱导的ARPE-19细胞衰老，并改善NaIO 3处理的hHTRA 1-Tg小鼠的视觉功能。我们的研究表明，升高的HTRA 1通过破坏线粒体功能和激活HIF-1信号通路促进RPE细胞衰老而参与AMD的发病机制。提示抑制HIF-1信号通路可能成为AMD潜在的治疗策略。视频摘要在线版本包含补充材料，可通过10. 1186/s12964-023-01138-9获取。
Age-related macular degeneration (AMD), characterized by the degeneration of retinal pigment epithelium (RPE) and photoreceptors, is the leading cause of irreversible vision impairment among the elderly. RPE senescence is an important contributor to AMD and has become a potential target for AMD therapy. HTRA1 is one of the most significant susceptibility genes in AMD, however, the correlation between HTRA1 and RPE senescence hasn’t been investigated in the pathogenesis of AMD. Western blotting and immunohistochemistry were used to detect HTRA1 expression in WT and transgenic mice overexpressing human HTRA1 (hHTRA1-Tg mice). RT-qPCR was used to detect the SASP in hHTRA1-Tg mice and ARPE-19 cells infected with HTRA1. TEM, SA-β-gal was used to detect the mitochondria and senescence in RPE. Retinal degeneration of mice was investigated by fundus photography, FFA, SD-OCT and ERG. The RNA-Seq dataset of ARPE-19 cells treated with adv-HTRA1 versus adv-NC were analyzed. Mitochondrial respiration and glycolytic capacity in ARPE-19 cells were measured using OCR and ECAR. Hypoxia of ARPE-19 cells was detected using EF5 Hypoxia Detection Kit. KC7F2 was used to reduce the HIF1α expression both in vitro and in vivo. In our study, we found that RPE senescence was facilitated in hHTRA1-Tg mice. And hHTRA1-Tg mice became more susceptible to NaIO3 in the development of oxidative stress-induced retinal degeneration. Similarly, overexpression of HTRA1 in ARPE-19 cells accelerated cellular senescence. Our RNA-seq revealed an overlap between HTRA1-induced differentially expressed genes associated with aging and those involved in mitochondrial function and hypoxia response in ARPE-19 cells. HTRA1 overexpression in ARPE-19 cells impaired mitochondrial function and augmented glycolytic capacity. Importantly, upregulation of HTRA1 remarkably activated HIF-1 signaling, shown as promoting HIF1α expression which mainly located in the nucleus. HIF1α translation inhibitor KC7F2 significantly prevented HTRA1-induced cellular senescence in ARPE-19 cells, as well as improved the visual function in hHTRA1-Tg mice treated with NaIO3. Our study showed elevated HTRA1 contributes to the pathogenesis of AMD by promoting cellular senescence in RPE through damaging mitochondrial function and activating HIF-1 signaling. It also pointed out that inhibition of HIF-1 signaling might serve as a potential therapeutic strategy for AMD. Video Abstract The online version contains supplementary material available at 10.1186/s12964-023-01138-9.
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影响因子：
3.7
作者：
Chen M;Hombrebueno JR;Luo C;Penalva R;Zhao J;Colhoun L;Pandi SP;Forrester JV;Xu H
通讯作者：
Xu H
影响因子：
3.6
作者：
Liguori I;Russo G;Curcio F;Bulli G;Aran L;Della-Morte D;Gargiulo G;Testa G;Cacciatore F;Bonaduce D;Abete P
通讯作者：
Abete P
影响因子：
14.8
作者：
Fernandez-Godino R;Garland DL;Pierce EA
通讯作者：
Pierce EA
影响因子：
17.8
作者：
Datta S;Cano M;Ebrahimi K;Wang L;Handa JT
通讯作者：
Handa JT
影响因子：
3.5
作者：
Chapman, James;Fielder, Edward;Passos, Joao F.
通讯作者：
Passos, Joao F.