Dot1(disruptor of telomeric silencing 1)最初在芽殖酵母中通过遗传筛选基因被发现,缺失则会引起端粒沉默。Dot1和其脊椎动物同源物DOT1L(Dot1-like)都具有催化组蛋白H3第79位赖氨酸(H3K79)甲基化的组蛋白甲基转移酶活性。在人类各种...Dot1(disruptor of telomeric silencing 1)最初在芽殖酵母中通过遗传筛选基因被发现,缺失则会引起端粒沉默。Dot1和其脊椎动物同源物DOT1L(Dot1-like)都具有催化组蛋白H3第79位赖氨酸(H3K79)甲基化的组蛋白甲基转移酶活性。在人类各种组织中,DOT1L基因启动子区域的CpG岛均呈现极低的DNA甲基化水平,说明DOT1L基因在人体中广泛中等程度表达。DOT1L基因在EBV转化的细胞及睾丸中表达水平升高。Dot1/DOT1L介导的H3K79甲基化参与多种生物学过程,并在小鼠基因损伤实验中发现DOT1L在心功能、红细胞生成、白血病及多种肿瘤的疾病发展中起重要的作用,因而DOT1L已成为针对表观遗传修饰因子的重要药物靶点。本文对Dot1/DOT1L在生理和病理方面的研究进展作一综述。展开更多
Recent research certified that DOT1L and its mutations represented by R231Q were potential targets for the treatment of lung cancer.Herein,a series of adenosine-containing derivatives were identified with DOT1LR231Q i...Recent research certified that DOT1L and its mutations represented by R231Q were potential targets for the treatment of lung cancer.Herein,a series of adenosine-containing derivatives were identified with DOT1LR231Q inhibition through antiproliferation assay and Western blot analysis in the H460R231Q cell.The most promising compound 37 significantly reduced DOT1LR231Q mediated H3K79 methylation and effectively inhibited the proliferation,self-renewal,migration,and invasion of lung cancer cell lines at low micromolar concentrations.The cell permeability and cellular target engagement of 37 were verified by both CETSA and DARTS assays.In the H460R231Q OE cell-derived xenograft(CDX)model,37 displayed pronounced tumor growth inhibition after intraperitoneal administration at 20 mg/kg dose for 3 weeks(TGI=54.38%),without obvious toxicities.A pharmacokinetic study revealed that 37 possessed tolerable properties(t_(1/2)=1.93±0.91 h,F=97.2%)after intraperitoneal administration in rats.Mechanism study confirmed that 37 suppressed malignant phenotypes of lung cancer carrying R231Q gain-of-function mutation via the MAPK/ERK signaling pathway.Moreover,analysis of the binding modes between molecules and DOT1LWT/R231Q proteins put forward the“Induced-fit”allosteric model in favor to the discovery of potent DOT1L candidates.展开更多
Epigenetic modification,including histone modification,precisely controls target gene expression.The posttranscriptional regulation of the innate signaling-triggered production of inflammatory cytokines and type I int...Epigenetic modification,including histone modification,precisely controls target gene expression.The posttranscriptional regulation of the innate signaling-triggered production of inflammatory cytokines and type I interferons has been fully elucidated,whereas the roles of histone modification alteration and epigenetic modifiers in regulating inflammatory responses need to be further explored.Di/tri-methylation modifications of histone 3 lysine 79(H3K79me2/3)have been shown to be associated with gene transcriptional activation.Disruptor of telomeric silencing-1-like(Dot1l)is the only known exclusive H3K79 methyltransferase and regulates the proliferation and differentiation of tumor cells.However,the roles of Dot1l and Dot1l-mediated H3K79 methylation in innate immunity and inflammatory responses remain unclear.Here,we found that H3K79me2/3 modification levels at the Il6 and Ifnb1 promoters,as well as H3K79me2 modification at the Tnfαpromoter,were increased in macrophages activated by Toll-like receptor(TLR)ligands or virus infection.The innate signals upregulated Dot1l expression in macrophages and THP1 cells.Dot1l silencing or a Dot1l inhibitor preferentially suppressed the production of IL-6 and interferon(IFN)-βbut not of TNFαin macrophages and THP1 cells triggered by TLR ligands or virus infection.Dot1l was recruited to the proximal promoter of the Il6 and Ifnb1 but not Tnfαgene and then mediated H3K79me2/3 modification at the Il6 and Ifnb1 promoters,consequently facilitating the transcription and expression of Il6 and Ifnb1.Thus,Dot1l-mediated selective H3K79me2/3 modifications at the Il6 and Ifnb1 promoters are required for the full activation of innate immune responses.This finding adds new insights into the epigenetic regulation of inflammatory responses and pathogenesis of autoimmune diseases.展开更多
急性白血病(AL)为克隆性恶性疾病,其主要特征表现为异常原始细胞大量增殖积聚,并不断对其他组织和器官进行浸润。研究证实,组蛋白对基因调控具有重要作用,现已发现的端粒沉默样阻断蛋白(DOT1L)是一种组蛋白,其缺乏Su(var)3-9,Enhancer-o...急性白血病(AL)为克隆性恶性疾病,其主要特征表现为异常原始细胞大量增殖积聚,并不断对其他组织和器官进行浸润。研究证实,组蛋白对基因调控具有重要作用,现已发现的端粒沉默样阻断蛋白(DOT1L)是一种组蛋白,其缺乏Su(var)3-9,Enhancer-of-zeste and Trithorax(SET)结构域,具有赖氨酸甲基转移酶(H-Lys-M)活性,可定向性促进组蛋白H3第79位赖氨酸(H3K79)的快速甲基化,这一过程与AL的发生与发展密切相关。本文根据DOT1L结构特点与功能特点,综合论述DOT1L促进H3K79的过度甲基化在AL的发生、发展及预后中的作用,借以彰显DOT1L在AL靶位研究和靶向治疗中的意义与应用前景。展开更多
文摘Dot1(disruptor of telomeric silencing 1)最初在芽殖酵母中通过遗传筛选基因被发现,缺失则会引起端粒沉默。Dot1和其脊椎动物同源物DOT1L(Dot1-like)都具有催化组蛋白H3第79位赖氨酸(H3K79)甲基化的组蛋白甲基转移酶活性。在人类各种组织中,DOT1L基因启动子区域的CpG岛均呈现极低的DNA甲基化水平,说明DOT1L基因在人体中广泛中等程度表达。DOT1L基因在EBV转化的细胞及睾丸中表达水平升高。Dot1/DOT1L介导的H3K79甲基化参与多种生物学过程,并在小鼠基因损伤实验中发现DOT1L在心功能、红细胞生成、白血病及多种肿瘤的疾病发展中起重要的作用,因而DOT1L已成为针对表观遗传修饰因子的重要药物靶点。本文对Dot1/DOT1L在生理和病理方面的研究进展作一综述。
基金supported by the National Natural Science Foundation of China(No.82173685 and No.82073320)Chinese Pharma-ceutical Association-Yi ling Biomedical Innovation Fund(No.CPAYLJ202001,China)+3 种基金Liao Ning Revitalization Talents Program(No.XLYC2002115,China)Key R&D Plan of Liaoning Province in 2020(No.2020020215-JH2/103,China)Development Project of Ministry of Education Innovation Team(No.IRT1073,China)Natural Science Foundation of Shenyang(22-315-6-11,China).
文摘Recent research certified that DOT1L and its mutations represented by R231Q were potential targets for the treatment of lung cancer.Herein,a series of adenosine-containing derivatives were identified with DOT1LR231Q inhibition through antiproliferation assay and Western blot analysis in the H460R231Q cell.The most promising compound 37 significantly reduced DOT1LR231Q mediated H3K79 methylation and effectively inhibited the proliferation,self-renewal,migration,and invasion of lung cancer cell lines at low micromolar concentrations.The cell permeability and cellular target engagement of 37 were verified by both CETSA and DARTS assays.In the H460R231Q OE cell-derived xenograft(CDX)model,37 displayed pronounced tumor growth inhibition after intraperitoneal administration at 20 mg/kg dose for 3 weeks(TGI=54.38%),without obvious toxicities.A pharmacokinetic study revealed that 37 possessed tolerable properties(t_(1/2)=1.93±0.91 h,F=97.2%)after intraperitoneal administration in rats.Mechanism study confirmed that 37 suppressed malignant phenotypes of lung cancer carrying R231Q gain-of-function mutation via the MAPK/ERK signaling pathway.Moreover,analysis of the binding modes between molecules and DOT1LWT/R231Q proteins put forward the“Induced-fit”allosteric model in favor to the discovery of potent DOT1L candidates.
基金This work was supported by the National Natural Science Foundation of China(81788101,31570871,31770970)CAMS Innovation Fund for Medical Sciences(2016-12M-1-003)the National Key Basic Research Program of China(2015CB964403).
文摘Epigenetic modification,including histone modification,precisely controls target gene expression.The posttranscriptional regulation of the innate signaling-triggered production of inflammatory cytokines and type I interferons has been fully elucidated,whereas the roles of histone modification alteration and epigenetic modifiers in regulating inflammatory responses need to be further explored.Di/tri-methylation modifications of histone 3 lysine 79(H3K79me2/3)have been shown to be associated with gene transcriptional activation.Disruptor of telomeric silencing-1-like(Dot1l)is the only known exclusive H3K79 methyltransferase and regulates the proliferation and differentiation of tumor cells.However,the roles of Dot1l and Dot1l-mediated H3K79 methylation in innate immunity and inflammatory responses remain unclear.Here,we found that H3K79me2/3 modification levels at the Il6 and Ifnb1 promoters,as well as H3K79me2 modification at the Tnfαpromoter,were increased in macrophages activated by Toll-like receptor(TLR)ligands or virus infection.The innate signals upregulated Dot1l expression in macrophages and THP1 cells.Dot1l silencing or a Dot1l inhibitor preferentially suppressed the production of IL-6 and interferon(IFN)-βbut not of TNFαin macrophages and THP1 cells triggered by TLR ligands or virus infection.Dot1l was recruited to the proximal promoter of the Il6 and Ifnb1 but not Tnfαgene and then mediated H3K79me2/3 modification at the Il6 and Ifnb1 promoters,consequently facilitating the transcription and expression of Il6 and Ifnb1.Thus,Dot1l-mediated selective H3K79me2/3 modifications at the Il6 and Ifnb1 promoters are required for the full activation of innate immune responses.This finding adds new insights into the epigenetic regulation of inflammatory responses and pathogenesis of autoimmune diseases.
文摘急性白血病(AL)为克隆性恶性疾病,其主要特征表现为异常原始细胞大量增殖积聚,并不断对其他组织和器官进行浸润。研究证实,组蛋白对基因调控具有重要作用,现已发现的端粒沉默样阻断蛋白(DOT1L)是一种组蛋白,其缺乏Su(var)3-9,Enhancer-of-zeste and Trithorax(SET)结构域,具有赖氨酸甲基转移酶(H-Lys-M)活性,可定向性促进组蛋白H3第79位赖氨酸(H3K79)的快速甲基化,这一过程与AL的发生与发展密切相关。本文根据DOT1L结构特点与功能特点,综合论述DOT1L促进H3K79的过度甲基化在AL的发生、发展及预后中的作用,借以彰显DOT1L在AL靶位研究和靶向治疗中的意义与应用前景。
