PDF(Pubmed)
Abstract：
UNASSIGNED: This study revealed a core regulator and common upstream mechanisms for the multifaceted pathological processes of age-related macular degeneration (AMD) and provided proof-of-concept for this new therapeutic target.
UNASSIGNED: Comprehensive gene expression analysis was performed using RNA sequencing of eye cup from old mice as well as laser-induced choroidal neovascularization (CNV) mouse model. Through integrative analysis and protein-protein interaction (PPI) analysis, common pathways and key transcription factor was identified simultaneously engaged in age-related retinal degeneration and CNV, the two typical pathological process of AMD. Subsequently, the expression changes of Spi1, the key regulator, as well as the alternation of the downstream mechanisms were validated in both models through qRT-PCR, Elisa, flow cytometry and immunofluorescence. Further, we assessed the impact of Spi1 knockdown in vitro and in vivo using gene intervention vectors carried by adeno-associated virus or lentivirus to test its potential as a therapeutic target.
UNASSIGNED: Compared to corresponding controls, we found 1,939 and 1,319 genes differentially expressed in eye cups of old and CNV mice respectively. The integrative analysis identified a total of 275 overlapping DEGs, of which 150 genes were co-upregulated. PPI analysis verified a central transcription factor, SPI1. The significant upregulation of Spi1 expression was then validated in both models, accompanied by macrophage polarization towards the M1 phenotype. Finally, SPI1 suppression significantly inhibited M1 polarization of BMDMs and attenuated neovascularization in CNV mice.
UNASSIGNED: This study demonstrates that SPI1 exerts a pivotal role in AMD by regulation of macrophage polarization and innate immune response, offering promise as an innovative target for treating AMD.
UNASSIGNED: Comprehensive gene expression analysis was performed using RNA sequencing of eye cup from old mice as well as laser-induced choroidal neovascularization (CNV) mouse model. Through integrative analysis and protein-protein interaction (PPI) analysis, common pathways and key transcription factor was identified simultaneously engaged in age-related retinal degeneration and CNV, the two typical pathological process of AMD. Subsequently, the expression changes of Spi1, the key regulator, as well as the alternation of the downstream mechanisms were validated in both models through qRT-PCR, Elisa, flow cytometry and immunofluorescence. Further, we assessed the impact of Spi1 knockdown in vitro and in vivo using gene intervention vectors carried by adeno-associated virus or lentivirus to test its potential as a therapeutic target.
UNASSIGNED: Compared to corresponding controls, we found 1,939 and 1,319 genes differentially expressed in eye cups of old and CNV mice respectively. The integrative analysis identified a total of 275 overlapping DEGs, of which 150 genes were co-upregulated. PPI analysis verified a central transcription factor, SPI1. The significant upregulation of Spi1 expression was then validated in both models, accompanied by macrophage polarization towards the M1 phenotype. Finally, SPI1 suppression significantly inhibited M1 polarization of BMDMs and attenuated neovascularization in CNV mice.
UNASSIGNED: This study demonstrates that SPI1 exerts a pivotal role in AMD by regulation of macrophage polarization and innate immune response, offering promise as an innovative target for treating AMD.
摘要：
■这项研究揭示了年龄相关性黄斑变性（AMD）多方面病理过程的核心调节因子和共同的上游机制，并为这一新的治疗靶点提供了概念验证。
使用来自老年小鼠的眼杯的RNA测序以及激光诱导的脉络膜新生血管形成(CNV)小鼠模型进行综合基因表达分析。通过整合分析和蛋白质-蛋白质相互作用(PPI)分析，发现了同时参与年龄相关性视网膜变性和CNV的共同通路和关键转录因子,AMD的两种典型病理过程。随后，关键调节剂Spi1的表达变化，以及通过qRT-PCR在两个模型中验证了下游机制的交替，Elisa,流式细胞术和免疫荧光。Further,我们使用腺相关病毒或慢病毒携带的基因干预载体在体外和体内评估了Spi1敲低的影响,以测试其作为治疗靶标的潜力.
■与相应的对照相比，我们发现1,939和1,319个基因分别在老年和CNV小鼠的眼杯中差异表达。综合分析确定了总共275个重叠的DEG，其中150个基因共同上调。PPI分析验证了一个中枢转录因子，SPI1.然后在两个模型中验证了Spi1表达的显着上调，伴随着巨噬细胞向M1表型的极化。最后,SPI1抑制显着抑制BMDMs的M1极化并减轻CNV小鼠的新生血管形成。
■这项研究表明，SPI1通过调节巨噬细胞极化和先天免疫反应在AMD中发挥关键作用，提供承诺作为治疗AMD的创新目标。
使用来自老年小鼠的眼杯的RNA测序以及激光诱导的脉络膜新生血管形成(CNV)小鼠模型进行综合基因表达分析。通过整合分析和蛋白质-蛋白质相互作用(PPI)分析，发现了同时参与年龄相关性视网膜变性和CNV的共同通路和关键转录因子,AMD的两种典型病理过程。随后，关键调节剂Spi1的表达变化，以及通过qRT-PCR在两个模型中验证了下游机制的交替，Elisa,流式细胞术和免疫荧光。Further,我们使用腺相关病毒或慢病毒携带的基因干预载体在体外和体内评估了Spi1敲低的影响,以测试其作为治疗靶标的潜力.
■与相应的对照相比，我们发现1,939和1,319个基因分别在老年和CNV小鼠的眼杯中差异表达。综合分析确定了总共275个重叠的DEG，其中150个基因共同上调。PPI分析验证了一个中枢转录因子，SPI1.然后在两个模型中验证了Spi1表达的显着上调，伴随着巨噬细胞向M1表型的极化。最后,SPI1抑制显着抑制BMDMs的M1极化并减轻CNV小鼠的新生血管形成。
■这项研究表明，SPI1通过调节巨噬细胞极化和先天免疫反应在AMD中发挥关键作用，提供承诺作为治疗AMD的创新目标。
