赖氨酸特异性去甲基化酶6B(lysine-specific demethylase 6B,KDM6B)是含Jumonji C结构域蛋白家族(Jumonji C domain-containing protein family,JmjC)中的一种重要表观遗传因子,不仅在细胞分化、炎症反应、组织稳态和神经性疾病中发挥...赖氨酸特异性去甲基化酶6B(lysine-specific demethylase 6B,KDM6B)是含Jumonji C结构域蛋白家族(Jumonji C domain-containing protein family,JmjC)中的一种重要表观遗传因子,不仅在细胞分化、炎症反应、组织稳态和神经性疾病中发挥表观遗传调控作用,还对巨噬细胞的功能、免疫反应等具有关键调控意义。作为JmjC家族中唯一能响应类Toll受体(Toll-like receptor,TLR)信号的成员,KDM6B可在TLR信号刺激下被激活从而发挥功能。研究发现,KDM6B可以通过调节巨噬细胞的极化、影响细胞因子的表达水平以及参与肿瘤微环境调控等方式影响巨噬细胞,因此,KDM6B在免疫反应、炎症反应以及肿瘤等病理生理过程中发挥重要作用。KDM6B作为关键的表观遗传因子对巨噬细胞的功能具有调控作用,包括调节巨噬细胞的极化、炎症反应以及促纤维化等,有望成为研究免疫、炎症及肿瘤等相关疾病的潜在靶点。展开更多
目的:筛选三阴性乳腺癌(triple-negative breast cancer,TNBC)潜在生物标志物并进行验证,探讨TNBC预后差、难治疗的遗传因素。方法:GEO与TCGA下载TNBC表达谱数据,使用GEO2R与R语言筛选差异表达基因(differentially expressed genes,DEGs...目的:筛选三阴性乳腺癌(triple-negative breast cancer,TNBC)潜在生物标志物并进行验证,探讨TNBC预后差、难治疗的遗传因素。方法:GEO与TCGA下载TNBC表达谱数据,使用GEO2R与R语言筛选差异表达基因(differentially expressed genes,DEGs);KEGG与GO富集分析;STRING数据库与Cytoscape软件绘制PPI网络并筛选核心基因;Kaplan-Meier数据库绘制生存曲线;利用bc-GenExMiner与UALCAN数据库验证以上结果;CIBERSORT算法与TIMER数据库分析预后相关基因对免疫细胞浸润及免疫检查点基因表达量的影响;qRT-PCR与Western Blot鉴定目的基因在TNBC与非三阴性乳腺癌(non-triple-negative breast cancer,n-TNBC)间的表达差异;si-RNA敲减目的基因;划痕实验与Transwell实验对比目的基因敲减与否对癌细胞侵袭、迁移的影响。结果:共得到123个TNBC相关DEGs,主要与雌激素信号通路相关;MCODE聚类分析筛选出16个核心基因;KRT6B、CDH3与预后相关,其中KRT6B在TNBC中高表达;KRT6B能够减少γδT细胞浸润,使γδT细胞表面CD27与PTPRC(CD45)减少;KRT6B表达量与CD276(B7-H3)表达量呈正相关;qRT-PCR与Western Blot结果显示KRT6B在TNBC中高表达;KRT6B经敲减后TNBC细胞侵袭、迁移能力均明显减弱。结论:KRT6B在TNBC中高表达,可能通过抑制γδT细胞浸润、减少γδT细胞表面标志物的表达并下调免疫检查点CD276的表达,与TNBC不良预后相关,有望成为诊断与治疗的新靶点。展开更多
Retinitis pigmentosa is a group of inherited diseases that lead to retinal degeneration and photoreceptor cell death.However,there is no effective treatment for retinitis pigmentosa caused by PDE6B mutation.Adeno-asso...Retinitis pigmentosa is a group of inherited diseases that lead to retinal degeneration and photoreceptor cell death.However,there is no effective treatment for retinitis pigmentosa caused by PDE6B mutation.Adeno-associated virus(AAV)-mediated gene therapy is a promising strategy for treating retinitis pigmentosa.The aim of this study was to explore the molecular mechanisms by which AAV2-PDE6B rescues retinal function.To do this,we injected retinal degeneration 10(rd10)mice subretinally with AAV2-PDE6B and assessed the therapeutic effects on retinal function and structure using dark-and light-adapted electroretinogram,optical coherence tomography,and immunofluorescence.Data-independent acquisition-mass spectrometry-based proteomic analysis was conducted to investigate protein expression levels and pathway enrichment,and the results from this analysis were verified by real-time polymerase chain reaction and western blotting.AAV2-PDE6B injection significantly upregulated PDE6βexpression,preserved electroretinogram responses,and preserved outer nuclear layer thickness in rd10 mice.Differentially expressed proteins between wild-type and rd10 mice were closely related to visual perception,and treating rd10 mice with AAV2-PDE6B restored differentially expressed protein expression to levels similar to those seen in wild-type mice.Kyoto Encyclopedia of Genes and Genome analysis showed that the differentially expressed proteins whose expression was most significantly altered by AAV2-PDE6B injection were enriched in phototransduction pathways.Furthermore,the phototransductionrelated proteins Pde6α,Rom1,Rho,Aldh1a1,and Rbp1 exhibited opposite expression patterns in rd10 mice with or without AAV2-PDE6B treatment.Finally,Bax/Bcl-2,p-ERK/ERK,and p-c-Fos/c-Fos expression levels decreased in rd10 mice following AAV2-PDE6B treatment.Our data suggest that AAV2-PDE6B-mediated gene therapy promotes phototransduction and inhibits apoptosis by inhibiting the ERK signaling pathway and upregulating Bcl-2/Bax expression in retinitis pigmentosa.展开更多
文摘赖氨酸特异性去甲基化酶6B(lysine-specific demethylase 6B,KDM6B)是含Jumonji C结构域蛋白家族(Jumonji C domain-containing protein family,JmjC)中的一种重要表观遗传因子,不仅在细胞分化、炎症反应、组织稳态和神经性疾病中发挥表观遗传调控作用,还对巨噬细胞的功能、免疫反应等具有关键调控意义。作为JmjC家族中唯一能响应类Toll受体(Toll-like receptor,TLR)信号的成员,KDM6B可在TLR信号刺激下被激活从而发挥功能。研究发现,KDM6B可以通过调节巨噬细胞的极化、影响细胞因子的表达水平以及参与肿瘤微环境调控等方式影响巨噬细胞,因此,KDM6B在免疫反应、炎症反应以及肿瘤等病理生理过程中发挥重要作用。KDM6B作为关键的表观遗传因子对巨噬细胞的功能具有调控作用,包括调节巨噬细胞的极化、炎症反应以及促纤维化等,有望成为研究免疫、炎症及肿瘤等相关疾病的潜在靶点。
文摘目的:筛选三阴性乳腺癌(triple-negative breast cancer,TNBC)潜在生物标志物并进行验证,探讨TNBC预后差、难治疗的遗传因素。方法:GEO与TCGA下载TNBC表达谱数据,使用GEO2R与R语言筛选差异表达基因(differentially expressed genes,DEGs);KEGG与GO富集分析;STRING数据库与Cytoscape软件绘制PPI网络并筛选核心基因;Kaplan-Meier数据库绘制生存曲线;利用bc-GenExMiner与UALCAN数据库验证以上结果;CIBERSORT算法与TIMER数据库分析预后相关基因对免疫细胞浸润及免疫检查点基因表达量的影响;qRT-PCR与Western Blot鉴定目的基因在TNBC与非三阴性乳腺癌(non-triple-negative breast cancer,n-TNBC)间的表达差异;si-RNA敲减目的基因;划痕实验与Transwell实验对比目的基因敲减与否对癌细胞侵袭、迁移的影响。结果:共得到123个TNBC相关DEGs,主要与雌激素信号通路相关;MCODE聚类分析筛选出16个核心基因;KRT6B、CDH3与预后相关,其中KRT6B在TNBC中高表达;KRT6B能够减少γδT细胞浸润,使γδT细胞表面CD27与PTPRC(CD45)减少;KRT6B表达量与CD276(B7-H3)表达量呈正相关;qRT-PCR与Western Blot结果显示KRT6B在TNBC中高表达;KRT6B经敲减后TNBC细胞侵袭、迁移能力均明显减弱。结论:KRT6B在TNBC中高表达,可能通过抑制γδT细胞浸润、减少γδT细胞表面标志物的表达并下调免疫检查点CD276的表达,与TNBC不良预后相关,有望成为诊断与治疗的新靶点。
基金supported by the National Natural Science Foundation of China,Nos.82071008(to BL)and 82004001(to XJ)Medical Science and Technology Program of Health Commission of Henan Province,No.LHGJ20210072(to RQ)Science and Technology Department of Henan Province,No.212102310307(to XJ)。
文摘Retinitis pigmentosa is a group of inherited diseases that lead to retinal degeneration and photoreceptor cell death.However,there is no effective treatment for retinitis pigmentosa caused by PDE6B mutation.Adeno-associated virus(AAV)-mediated gene therapy is a promising strategy for treating retinitis pigmentosa.The aim of this study was to explore the molecular mechanisms by which AAV2-PDE6B rescues retinal function.To do this,we injected retinal degeneration 10(rd10)mice subretinally with AAV2-PDE6B and assessed the therapeutic effects on retinal function and structure using dark-and light-adapted electroretinogram,optical coherence tomography,and immunofluorescence.Data-independent acquisition-mass spectrometry-based proteomic analysis was conducted to investigate protein expression levels and pathway enrichment,and the results from this analysis were verified by real-time polymerase chain reaction and western blotting.AAV2-PDE6B injection significantly upregulated PDE6βexpression,preserved electroretinogram responses,and preserved outer nuclear layer thickness in rd10 mice.Differentially expressed proteins between wild-type and rd10 mice were closely related to visual perception,and treating rd10 mice with AAV2-PDE6B restored differentially expressed protein expression to levels similar to those seen in wild-type mice.Kyoto Encyclopedia of Genes and Genome analysis showed that the differentially expressed proteins whose expression was most significantly altered by AAV2-PDE6B injection were enriched in phototransduction pathways.Furthermore,the phototransductionrelated proteins Pde6α,Rom1,Rho,Aldh1a1,and Rbp1 exhibited opposite expression patterns in rd10 mice with or without AAV2-PDE6B treatment.Finally,Bax/Bcl-2,p-ERK/ERK,and p-c-Fos/c-Fos expression levels decreased in rd10 mice following AAV2-PDE6B treatment.Our data suggest that AAV2-PDE6B-mediated gene therapy promotes phototransduction and inhibits apoptosis by inhibiting the ERK signaling pathway and upregulating Bcl-2/Bax expression in retinitis pigmentosa.
