【目的】克隆白来航鸡干扰素基因刺激因子基因(stimulator of interferon genes,STING)CDS区序列并进行生物信息学和组织表达分析,为阐明STING基因在抗病毒免疫应答中的作用奠定基础。【方法】采用PCR扩增并克隆白来航鸡STING基因CDS区...【目的】克隆白来航鸡干扰素基因刺激因子基因(stimulator of interferon genes,STING)CDS区序列并进行生物信息学和组织表达分析,为阐明STING基因在抗病毒免疫应答中的作用奠定基础。【方法】采用PCR扩增并克隆白来航鸡STING基因CDS区,测序后对其编码氨基酸序列进行相似性比对及系统进化树构建,利用生物信息学预测STING蛋白的理化特性及结构功能,并利用实时荧光定量PCR技术检测STING基因在鸡心脏、肝脏等14个组织中的表达情况。【结果】白来航鸡STING基因CDS区序列全长1140 bp,编码379个氨基酸。相似性比对和系统进化树分析结果表明,白来航鸡STING基因与原鸡的相似性最高(99.7%),亲缘关系最近,与冠小嘴乌鸦亲缘关系最远。STING蛋白为酸性、亲水性蛋白,分子质量为42.625 ku,等电点(pI)为6.67,不稳定系数为69.26,脂肪系数为105.01。该蛋白大部分在线粒体和内质网上合成,含有跨膜结构,不含信号肽。STING蛋白二级结构包括α-螺旋(54.62%)、延伸链(10.29%)、β-转角(3.43%)及无规则卷曲(31.66%)。蛋白互作分析表明,白来航鸡STING蛋白与NFKB1、DDX41、cGAS、TBK1等蛋白存在相互作用。实时荧光定量PCR结果显示,STING基因在白来航鸡组织中广泛表达,其中在肺脏中表达量最高,且显著高于其他组织(P<0.05);在胸肌中表达量最低。【结论】本研究成功克隆了白来航鸡STING基因,其CDS区序列全长1140 bp,编码379个氨基酸。白来航鸡STING蛋白为酸性、亲水性蛋白,含有跨膜结构。STING基因在白来航鸡肺脏中表达量最高。研究结果为深入探究白来航鸡STING基因编码蛋白功能提供了材料。展开更多
The intricate interplay between the human immune system and cancer development underscores the central role of immunotherapy in cancer treatment.Within this landscape,the innate immune system,a critical sentinel prote...The intricate interplay between the human immune system and cancer development underscores the central role of immunotherapy in cancer treatment.Within this landscape,the innate immune system,a critical sentinel protecting against tumor incursion,is a key player.The cyclic GMP-AMP synthase(c GAS)and stimulator of interferon genes(STING)pathway has been found to be a linchpin of innate immunity:activation of this signaling pathway orchestrates the production of type I interferon(IFN-α/β),thus fostering the maturation,differentiation,and mobilization of immune effectors in the tumor microenvironment.Furthermore,STING activation facilitates the release and presentation of tumor antigens,and therefore is an attractive target for cancer immunotherapy.Current strategies to activate the STING pathway,including use of pharmacological agonists,have made substantial advancements,particularly when combined with immune checkpoint inhibitors.These approaches have shown promise in preclinical and clinical settings,by enhancing patient survival rates.This review describes the evolving understanding of the c GAS-STING pathway's involvement in tumor biology and therapy.Moreover,this review explores classical and non-classical STING agonists,providing insights into their mechanisms of action and potential for optimizing immunotherapy strategies.Despite challenges and complexities,the c GAS-STING pathway,a promising avenue for enhancing cancer treatment efficacy,has the potential to revolutionize patient outcomes.展开更多
基金the National Key Research and Development Program of China(Grant Nos.2022YFC3401500 and 2020YFA0803201 to P.W.,and 2021YFA1302200 to L.F.)the National Natural Science Foundation of China(Grant Nos.31830053,31920103007,and 82341028 to P.W.+1 种基金82122056,82073153,and 31871398 to L.F.and 31900568 to P.W.)the Natural Science Foundation of Shanghai(Grant No.22ZR1450700 to Z.J.W.)。
文摘The intricate interplay between the human immune system and cancer development underscores the central role of immunotherapy in cancer treatment.Within this landscape,the innate immune system,a critical sentinel protecting against tumor incursion,is a key player.The cyclic GMP-AMP synthase(c GAS)and stimulator of interferon genes(STING)pathway has been found to be a linchpin of innate immunity:activation of this signaling pathway orchestrates the production of type I interferon(IFN-α/β),thus fostering the maturation,differentiation,and mobilization of immune effectors in the tumor microenvironment.Furthermore,STING activation facilitates the release and presentation of tumor antigens,and therefore is an attractive target for cancer immunotherapy.Current strategies to activate the STING pathway,including use of pharmacological agonists,have made substantial advancements,particularly when combined with immune checkpoint inhibitors.These approaches have shown promise in preclinical and clinical settings,by enhancing patient survival rates.This review describes the evolving understanding of the c GAS-STING pathway's involvement in tumor biology and therapy.Moreover,this review explores classical and non-classical STING agonists,providing insights into their mechanisms of action and potential for optimizing immunotherapy strategies.Despite challenges and complexities,the c GAS-STING pathway,a promising avenue for enhancing cancer treatment efficacy,has the potential to revolutionize patient outcomes.
