目的:探究尿液细胞外囊泡中关键蛋白细胞骨架相关蛋白4(cytoskeleton⁃associated protein 4,CKAP4)作为生物标志物预测糖尿病肾病(diabetic kidney disease,DKD)进展的能力。方法:纳入南京医科大学第一附属医院143例肾活检证实DKD的2型...目的:探究尿液细胞外囊泡中关键蛋白细胞骨架相关蛋白4(cytoskeleton⁃associated protein 4,CKAP4)作为生物标志物预测糖尿病肾病(diabetic kidney disease,DKD)进展的能力。方法:纳入南京医科大学第一附属医院143例肾活检证实DKD的2型糖尿病患者以及肾脏恶性肿瘤患者10例。免疫组化法检测肾组织中CKAP4的表达水平。采用Spearman相关分析CKAP4表达水平与临床指标的相关性。计算受试者工作特征(receiver operating characteristic,ROC)曲线下面积(area under the curve,AUC),以评估CKAP4表达是否能有效区分肾脏预后不良的患者。采用ROC曲线分析确定预测肾脏事件的CKAP4评分的最佳临界值(最高约登指数)。此外,进行时间依赖性AUC分析,以表征肾活检后6个月以上CKAP4的预测准确性。通过Cox比例风险模型随访CKAP4和DKD进展之间的风险比。将单变量分析中有统计学意义(P<0.05)的临床病理因素作为协变量纳入多变量Cox比例风险模型分析。采用Kaplan⁃Meier分析评价CKAP4高表达组和CKAP4低表达组肾活检后6个月以上生存的差异。结果:与肿瘤患者癌旁正常肾脏组织相比,CKAP4在DKD患者肾脏组织中表达增高,差异有统计学意义(P<0.001)。CKAP4在不同分期的DKD患者肾组织中表达有差异,其中Ⅱ期与Ⅲ期、Ⅱ期与Ⅳ期、Ⅲ期与Ⅳ期之间的差异均有统计学意义(P均<0.05)。DKD患者CKAP4表达与血清肌酐、尿素氮、24 h尿蛋白呈正相关,与肾小球滤过率估计值、血红蛋白呈负相关。DKD患者中位随访期为2.22年,63例(44.06%)患者出现DKD进展。Pearson相关分析显示,CKAP4表达水平随DKD病理分级的升高而升高(r=0.808,P<0.001)。多因素Cox回归分析显示,CKAP4高表达与DKD进展的风险增加相关(HR=4.120,95%CI:1.730~9.811,P=0.001)。此外,在Kaplan⁃Meier生存分析中,CKAP4高表达组患者的肾脏终点事件的发生率显著高于CKAP4低表达组(P<0.001)。绘制了包括CKAP4分类在内的列线图来预测DKD进展风险(C⁃index:0.689)。结论:来源于尿液细胞外囊泡的CKAP4是DKD患者肾活检后6个月以上疾病进展的独立危险因素。展开更多
细胞骨架相关蛋白4(cytoskeleton-associated protein 4,CKAP4)即p63,是一种非糖基化的Ⅱ型膜蛋白。CKAP4在组织、细胞以及血清中的表达可作为多种肿瘤的诊断性标志物,并与这些肿瘤的预后有一定相关性。该文对CKAP4的亚型和功能,以及CK...细胞骨架相关蛋白4(cytoskeleton-associated protein 4,CKAP4)即p63,是一种非糖基化的Ⅱ型膜蛋白。CKAP4在组织、细胞以及血清中的表达可作为多种肿瘤的诊断性标志物,并与这些肿瘤的预后有一定相关性。该文对CKAP4的亚型和功能,以及CKAP4作为肿瘤诊断性、预后性指标的研究及应用进行了综述。展开更多
Background:Since boosting stem cell resilience in stressful environments is critical for the therapeutic efficacy of stem cell-based transplantations in liver disease,this study aimed to establish the efficacy of a tr...Background:Since boosting stem cell resilience in stressful environments is critical for the therapeutic efficacy of stem cell-based transplantations in liver disease,this study aimed to establish the efficacy of a transient plasmid-based preconditioning strategy for boosting the capability of mesenchymal stromal cells(MSCs)for anti-inflammation/antioxidant defenses and paracrine actions in recipient hepatocytes.Methods:Human adipose mesenchymal stem cells(hADMSCs)were subjected to transfer,either with or without the nuclear factor erythroid 2-related factor 2(Nrf2)/Dickkopf1(DKK1)genes,followed by exposure to TNF-a/H2O2.Mouse models were subjected to acute chronic liver failure(ACLF)and subsequently injected with either transfected or untransfected MSCs.These hADMSCs and ACLF mouse models were used to investigate the interaction between Nrf2/DKK1 and the hepatocyte receptor cytoskeleton-associated protein 4(CKAP4).Results:Activation of Nrf2 and DKK1 enhanced the anti-stress capacity of MSCs in vitro.In a murine model of ACLF,transient co-overexpression of Nrf2 and DKK1 via plasmid transfection improved MSC resilience against inflammatory and oxidative assaults,boosted MSC transplantation efficacy,and promoted recipient liver regeneration due to a shift from the activation of the anti-regenerative IFN-c/STAT1 pathway to the pro-regenerative IL-6/STAT3 pathway in the liver.Importantly,the therapeutic benefits of MSC transplantation were nullified when the receptor CKAP4,which interacts with DKK1,was specifically removed from recipient hepatocytes.However,the removal of the another receptor low-density lipoprotein receptor-related protein 6(LRP6)had no impact on the effectiveness of MSC transplantation.Moreover,in long-term observations,no tumorigenicity was detected in mice following transplantation of transiently preconditioned MSCs.Conclusions:Co-stimulation with Nrf2/DKK1 safely improved the efficacy of human MSC-based therapies in murine models of ACLF through CKAP4-dependent paracrine mechanisms.展开更多
文摘目的:探究尿液细胞外囊泡中关键蛋白细胞骨架相关蛋白4(cytoskeleton⁃associated protein 4,CKAP4)作为生物标志物预测糖尿病肾病(diabetic kidney disease,DKD)进展的能力。方法:纳入南京医科大学第一附属医院143例肾活检证实DKD的2型糖尿病患者以及肾脏恶性肿瘤患者10例。免疫组化法检测肾组织中CKAP4的表达水平。采用Spearman相关分析CKAP4表达水平与临床指标的相关性。计算受试者工作特征(receiver operating characteristic,ROC)曲线下面积(area under the curve,AUC),以评估CKAP4表达是否能有效区分肾脏预后不良的患者。采用ROC曲线分析确定预测肾脏事件的CKAP4评分的最佳临界值(最高约登指数)。此外,进行时间依赖性AUC分析,以表征肾活检后6个月以上CKAP4的预测准确性。通过Cox比例风险模型随访CKAP4和DKD进展之间的风险比。将单变量分析中有统计学意义(P<0.05)的临床病理因素作为协变量纳入多变量Cox比例风险模型分析。采用Kaplan⁃Meier分析评价CKAP4高表达组和CKAP4低表达组肾活检后6个月以上生存的差异。结果:与肿瘤患者癌旁正常肾脏组织相比,CKAP4在DKD患者肾脏组织中表达增高,差异有统计学意义(P<0.001)。CKAP4在不同分期的DKD患者肾组织中表达有差异,其中Ⅱ期与Ⅲ期、Ⅱ期与Ⅳ期、Ⅲ期与Ⅳ期之间的差异均有统计学意义(P均<0.05)。DKD患者CKAP4表达与血清肌酐、尿素氮、24 h尿蛋白呈正相关,与肾小球滤过率估计值、血红蛋白呈负相关。DKD患者中位随访期为2.22年,63例(44.06%)患者出现DKD进展。Pearson相关分析显示,CKAP4表达水平随DKD病理分级的升高而升高(r=0.808,P<0.001)。多因素Cox回归分析显示,CKAP4高表达与DKD进展的风险增加相关(HR=4.120,95%CI:1.730~9.811,P=0.001)。此外,在Kaplan⁃Meier生存分析中,CKAP4高表达组患者的肾脏终点事件的发生率显著高于CKAP4低表达组(P<0.001)。绘制了包括CKAP4分类在内的列线图来预测DKD进展风险(C⁃index:0.689)。结论:来源于尿液细胞外囊泡的CKAP4是DKD患者肾活检后6个月以上疾病进展的独立危险因素。
文摘细胞骨架相关蛋白4(cytoskeleton-associated protein 4,CKAP4)即p63,是一种非糖基化的Ⅱ型膜蛋白。CKAP4在组织、细胞以及血清中的表达可作为多种肿瘤的诊断性标志物,并与这些肿瘤的预后有一定相关性。该文对CKAP4的亚型和功能,以及CKAP4作为肿瘤诊断性、预后性指标的研究及应用进行了综述。
基金funded by grants from the National Natural Science Foundation of China[nos 82122009,81970515,82170605,and 81873573]the Guangdong Natural Science Funds for Distinguished Young Scholar[no.2019B151502013]the Guangdong Basic and Applied Research Foundation[no.2021B1515120069].
文摘Background:Since boosting stem cell resilience in stressful environments is critical for the therapeutic efficacy of stem cell-based transplantations in liver disease,this study aimed to establish the efficacy of a transient plasmid-based preconditioning strategy for boosting the capability of mesenchymal stromal cells(MSCs)for anti-inflammation/antioxidant defenses and paracrine actions in recipient hepatocytes.Methods:Human adipose mesenchymal stem cells(hADMSCs)were subjected to transfer,either with or without the nuclear factor erythroid 2-related factor 2(Nrf2)/Dickkopf1(DKK1)genes,followed by exposure to TNF-a/H2O2.Mouse models were subjected to acute chronic liver failure(ACLF)and subsequently injected with either transfected or untransfected MSCs.These hADMSCs and ACLF mouse models were used to investigate the interaction between Nrf2/DKK1 and the hepatocyte receptor cytoskeleton-associated protein 4(CKAP4).Results:Activation of Nrf2 and DKK1 enhanced the anti-stress capacity of MSCs in vitro.In a murine model of ACLF,transient co-overexpression of Nrf2 and DKK1 via plasmid transfection improved MSC resilience against inflammatory and oxidative assaults,boosted MSC transplantation efficacy,and promoted recipient liver regeneration due to a shift from the activation of the anti-regenerative IFN-c/STAT1 pathway to the pro-regenerative IL-6/STAT3 pathway in the liver.Importantly,the therapeutic benefits of MSC transplantation were nullified when the receptor CKAP4,which interacts with DKK1,was specifically removed from recipient hepatocytes.However,the removal of the another receptor low-density lipoprotein receptor-related protein 6(LRP6)had no impact on the effectiveness of MSC transplantation.Moreover,in long-term observations,no tumorigenicity was detected in mice following transplantation of transiently preconditioned MSCs.Conclusions:Co-stimulation with Nrf2/DKK1 safely improved the efficacy of human MSC-based therapies in murine models of ACLF through CKAP4-dependent paracrine mechanisms.
