Although the exact etiology of inflammatory bowel disease(IBD)remains unclear,exaggerated immune response in genetically predisposed individuals has been reported.Th1 and Th17 cells mediate IBD development.Macrophages...Although the exact etiology of inflammatory bowel disease(IBD)remains unclear,exaggerated immune response in genetically predisposed individuals has been reported.Th1 and Th17 cells mediate IBD development.Macrophages produce IL-12 and IL-23 that share p40 subunit encoded by IL12B gene as heteromer partner to drive Th1 and Th17 differentiation.The available animal and human data strongly support the pathogenic role of IL-12/IL-23 in IBD development and suggest that blocking p40 might be the potential strategy for IBD treatment.Furthermore,aberrant alteration of some cytokines expression via epigenetic mechanisms is involved in pathogenesis o f IBD.In this study,we analyzed core promoter region of IL12B gene and investigated whether IL12B expression could be regulated through targeted epigenetic modification with gene editing technology.Transcription activator-like effectors(TALEs)are widely used in the field of genome editing and can specifically target DNA sequence in the host genome.We synthesized the TALE DNA-binding domains that target the promoter of human IL12B gene and fused it with the functional catalytic domains of epigenetic enzymes.Transient expression of these engineered enzymes demonstrated that the TALE-DNMT3A targeted the selected IL12B promoter region,induced loci-specific DNA methylation,and down-regulated IL-12B expression in various human cell lines.Collectively,our data suggested that epigenetic editing of IL12B through methylating DNA on its promoter might be developed as a potential therapeutic strategy for IBD treatment.展开更多
目的:利用生信分析探讨细胞因子IL-12及IFN-γ对乳腺癌患者免疫微环境的影响,并结合临床病理特征,对其评估新辅助化疗疗效的价值进行研究。方法:在基因表达综合(GEO)数据库中下载GSE163882,比较IL-12相对应基因IL12B及IFN-γ相对应基因I...目的:利用生信分析探讨细胞因子IL-12及IFN-γ对乳腺癌患者免疫微环境的影响,并结合临床病理特征,对其评估新辅助化疗疗效的价值进行研究。方法:在基因表达综合(GEO)数据库中下载GSE163882,比较IL-12相对应基因IL12B及IFN-γ相对应基因IFNG在病理完全缓解(pCR)患者和残留病灶(RD)患者中的表达。收集于我院接受新辅助化疗的50例乳腺癌患者的临床资料,使用卡方检验分析临床病理特征与新辅助化疗疗效的相关性;通过ROC曲线评估血清IL-12、IFN-γ的预测价值,并且确定最佳截断值;采用二元Logistic回归进行多因素分析。通过ssGSEA免疫浸润分析IL12B和IFNG与免疫细胞浸润的相关性;采用GO和KEGG功能富集分析,探讨相关基因潜在生物学功能及信号通路。结果:在GEO数据库GSE163882数据集中IL12B和IFNG在pCR患者中呈现高表达。于我院收治的临床患者中pCR患者化疗前血清IL-12和IFN-γ水平明显高于非pCR组(P γ表达状态显著相关(P γ水平是新辅助化疗疗效的独立影响因素。免疫浸润分析显示,活化CD4 T细胞、活化CD8 T细胞、活化B细胞和自然杀伤细胞在pCR患者中高度浸润。结论:血清IL-12和IFN-γ表达状态与新辅助化疗效果具有相关性,化疗前血清IFN-γ高表达是pCR的独立预测因子。Objective: To investigate the effects of cytokines IL-12 and IFN-γ on the immune microenvironment of breast cancer patients by means of Bioinformatics analysis, and to study their value in evaluating the efficacy of neoadjuvant chemotherapy combined with clinicopathological characteristics. Methods: GSE163882 was downloaded from the Gene Expression Omnibus database, and the expressions of IL-12 corresponding gene IL12B and IFNG corresponding gene IFN-γ were compared in pathologic complete response patients and residual lesion patients. The clinical data of 50 breast cancer patients who received neoadjuvant chemotherapy at The Affiliated Hospital of Qingdao University were collected. Chi-square test was used to analyze the correlation between clinicopathological features and the efficacy of neoadjuvant chemotherapy. The predictive value of serum IL-12 and IFN-γ was evaluated by ROC curve. Multivariate analysis was performed by binary Logistic regression. The correlation between IL12B and IFNG and immune cell infiltration was analyzed by ssGSEA immunoinfiltration. GO and KEGG functional enrichment analysis were used to explore the potential biological functions and signaling pathways of related genes. Results: IL12B and IFNG were highly expressed in pCR patients in the GSE163882 dataset. The serum levels of IL-12 and IFN-γ in pCR patients before chemotherapy were significantly higher than those in non-PCR group (P γ expression after neoadjuvant chemotherapy (P γ level before chemotherapy was an independent factor influencing the efficacy of neoadjuvant chemotherapy. Immunoinfiltration analysis showed that activated CD4 T cells, activated CD8 T cells, activated B cells, and natural killer cells were highly infiltrated in pCR patients. Conclusion: The expression of serum IL-12 and IFN-γ is correlated with the effect of neoadjuvant chemotherapy, and the high expression of serum IFN-γ before chemotherapy is an independent predictor of pCR.展开更多
基金The study was supported by the National Natural Science Foundation of China(No.81270468).
文摘Although the exact etiology of inflammatory bowel disease(IBD)remains unclear,exaggerated immune response in genetically predisposed individuals has been reported.Th1 and Th17 cells mediate IBD development.Macrophages produce IL-12 and IL-23 that share p40 subunit encoded by IL12B gene as heteromer partner to drive Th1 and Th17 differentiation.The available animal and human data strongly support the pathogenic role of IL-12/IL-23 in IBD development and suggest that blocking p40 might be the potential strategy for IBD treatment.Furthermore,aberrant alteration of some cytokines expression via epigenetic mechanisms is involved in pathogenesis o f IBD.In this study,we analyzed core promoter region of IL12B gene and investigated whether IL12B expression could be regulated through targeted epigenetic modification with gene editing technology.Transcription activator-like effectors(TALEs)are widely used in the field of genome editing and can specifically target DNA sequence in the host genome.We synthesized the TALE DNA-binding domains that target the promoter of human IL12B gene and fused it with the functional catalytic domains of epigenetic enzymes.Transient expression of these engineered enzymes demonstrated that the TALE-DNMT3A targeted the selected IL12B promoter region,induced loci-specific DNA methylation,and down-regulated IL-12B expression in various human cell lines.Collectively,our data suggested that epigenetic editing of IL12B through methylating DNA on its promoter might be developed as a potential therapeutic strategy for IBD treatment.
文摘目的:利用生信分析探讨细胞因子IL-12及IFN-γ对乳腺癌患者免疫微环境的影响,并结合临床病理特征,对其评估新辅助化疗疗效的价值进行研究。方法:在基因表达综合(GEO)数据库中下载GSE163882,比较IL-12相对应基因IL12B及IFN-γ相对应基因IFNG在病理完全缓解(pCR)患者和残留病灶(RD)患者中的表达。收集于我院接受新辅助化疗的50例乳腺癌患者的临床资料,使用卡方检验分析临床病理特征与新辅助化疗疗效的相关性;通过ROC曲线评估血清IL-12、IFN-γ的预测价值,并且确定最佳截断值;采用二元Logistic回归进行多因素分析。通过ssGSEA免疫浸润分析IL12B和IFNG与免疫细胞浸润的相关性;采用GO和KEGG功能富集分析,探讨相关基因潜在生物学功能及信号通路。结果:在GEO数据库GSE163882数据集中IL12B和IFNG在pCR患者中呈现高表达。于我院收治的临床患者中pCR患者化疗前血清IL-12和IFN-γ水平明显高于非pCR组(P γ表达状态显著相关(P γ水平是新辅助化疗疗效的独立影响因素。免疫浸润分析显示,活化CD4 T细胞、活化CD8 T细胞、活化B细胞和自然杀伤细胞在pCR患者中高度浸润。结论:血清IL-12和IFN-γ表达状态与新辅助化疗效果具有相关性,化疗前血清IFN-γ高表达是pCR的独立预测因子。Objective: To investigate the effects of cytokines IL-12 and IFN-γ on the immune microenvironment of breast cancer patients by means of Bioinformatics analysis, and to study their value in evaluating the efficacy of neoadjuvant chemotherapy combined with clinicopathological characteristics. Methods: GSE163882 was downloaded from the Gene Expression Omnibus database, and the expressions of IL-12 corresponding gene IL12B and IFNG corresponding gene IFN-γ were compared in pathologic complete response patients and residual lesion patients. The clinical data of 50 breast cancer patients who received neoadjuvant chemotherapy at The Affiliated Hospital of Qingdao University were collected. Chi-square test was used to analyze the correlation between clinicopathological features and the efficacy of neoadjuvant chemotherapy. The predictive value of serum IL-12 and IFN-γ was evaluated by ROC curve. Multivariate analysis was performed by binary Logistic regression. The correlation between IL12B and IFNG and immune cell infiltration was analyzed by ssGSEA immunoinfiltration. GO and KEGG functional enrichment analysis were used to explore the potential biological functions and signaling pathways of related genes. Results: IL12B and IFNG were highly expressed in pCR patients in the GSE163882 dataset. The serum levels of IL-12 and IFN-γ in pCR patients before chemotherapy were significantly higher than those in non-PCR group (P γ expression after neoadjuvant chemotherapy (P γ level before chemotherapy was an independent factor influencing the efficacy of neoadjuvant chemotherapy. Immunoinfiltration analysis showed that activated CD4 T cells, activated CD8 T cells, activated B cells, and natural killer cells were highly infiltrated in pCR patients. Conclusion: The expression of serum IL-12 and IFN-γ is correlated with the effect of neoadjuvant chemotherapy, and the high expression of serum IFN-γ before chemotherapy is an independent predictor of pCR.
