[AASLD 2012]模仿干扰素-α（IFNα）对乙型肝炎病毒（HBV）的转

肝胆速递

模仿干扰素-α（IFNα）对乙型肝炎病毒（HBV）的转录和复制的抑制活性:使用后生(epigenetic)的小分子针对核cccDNA的微小染色体的遗传控制(epigenetic control)
CONTROL ID: 1426150
PRESENTATION TYPE: Oral or Poster
CURRENT CATEGORY: Hepatitis B
CURRENT DESCRIPTORS: I02. Treatment and Clinical Trials
TITLE: Mimicking Interferon-α (IFNα) inhibitory activity on hepatitis B virus (HBV) transcription and replication by targeting the epigenetic control of nuclear cccDNA minichromosome with epigenetic small molecules
AUTHORS (FIRST NAME, LAST NAME): Laura Belloni1, 2, Gianna Aurora Palumbo1, 3, Sergio Valente4, Dante Rotili4, Natalia Pediconi1, 3, Antonello Mai4, Massimo Levrero1, 2
Institutional Author(s):
INSTITUTIONS (ALL): 1. Dept of Internal Medicine, Sapienza University, Rome, Italy.
2. Life Nanosciences Laboratory , Sapienza University , Rome, Italy.
3. EAL Inserm U785 , Sapienza University , Rome, Italy.
4. Dip. di Chimica e Tecnologie del Farmaco , Sapienza University , Rome, Italy.
ABSTRACT BODY: Background: HBV cccDNA, the nuclear replicative intermediate of HBV and template for all viral messenger RNAs, is organized into mini-chromosomes in the nucleus of infected cells by histone and non-histone proteins. By using a cccDNA-specific chromatin immunoprecipitation (ChIP)-based assay [that allows to study the impact of viral and cellular proteins on cccDNA mini-chromosome epigenetic modifications, chromatin dynamics, and transcriptional activity], we established that HBV replication is regulated, both in a cell replication system and in the liver of HBV chronically infected patients, by the acetylation status of cccDNA-bound H3/H4 histones. We have also shown that interferon-α (IFNα) inhibits HBV transcription and replication in vitro and in vivo by favoring the long term recruitment to the nuclear cccDNA mini-chromosome of the class III HDAC hSirt1 and of the PRC2 repressive complex, including the transcriptional co-repressors HDAC1 and Ezh2.
Aim: We sought to assess the feasibility to mimic IFNα repressive activity on HBV transcription and replication by targeting the epigenetic control of nuclear cccDNA mini-chromosome with small compounds active on different classes of chromatin modifying enzymes.
Methods: Capsid-associated HBV-DNA (TaqMan real-time PCR), cccDNA (TaqMan real-time PCR) and pgRNA levels (quantitative real-time PCR with specific primers), were assessed in HepG2 cells transfected with full length HBV genomes left untreated and or treated with a) class I/II and class III histone deacetylase inhibitors (HDACi); b) p300 and PCAF histone acetyltransferases (HAT) inhibitors; c) hSirt1 activators and d) JMJD3 histone demethylase inhibitors.
Results: We confirmed that class I/II HDACi potentiate cccDNA-bound histones acetylation, cccDNA transcription and HBV replication. Conversely, the combined inhibition of p300 and PCAF HATs (compound EML-264) resulted in an evident reduction of HBV replication that mirrored the decrease of pgRNA transcription. The hSirt1/2 activators MC2562 and MC2791, albeit with different efficiency, both inhibited HBV replication and cccDNA transcription. Potentiation of Ezh2 activity through the inhibition of JMJD3 histone demethylase with compound MC3119 resulted in a >50% reduction of pgRNA transcription. Notably, inhibition of hSirt1/2 (MC2344) or Ezh2 (MC2887) strongly induced cell death, thus hampering the evaluation of their effects on viral replication.
Conclusions: Altogether these results represent a proof of concept that activation of hSirt1 and Ezh2 by small molecules can induce an “active epigenetic suppression” of HBV cccDNA minichromosome similar to that observed with IFNα.

肝胆速递

标题：模仿干扰素-α（IFNα）对乙型肝炎病毒（HBV）的转录和复制的抑制活性与后生的小分子针对核cccDNA的微小染色体的表观遗传调控的
（FIRST NAME，LAST NAME）：劳拉Belloni1号，2，3，吉安娜极光Palumbo1，塞尔吉奥Valente4，但丁Rotili4，纳塔利娅Pediconi1，3，Mai4安东内洛，马西莫Levrero1，2
机构（S）：
机构（ALL）：1。内科部，Sapienza大学，罗马，意大利。
2。生命纳米科学实验室，Sapienza大学，罗马，意大利。
3。 EAL INSERM的U785，Sapienza大学，罗马，意大利。
4。 DIP封装。二化学ËTecnologie德尔Farmaco，Sapienza大学，罗马，意大利。
抽象的身体：背景：乙肝病毒cccDNA的，核的HBV复制中间体和病毒信使RNA模板为所有受感染的细胞的细胞核，分为迷你染色体组蛋白和非组蛋白。通过cccDNA的，特定的染色质免疫沉淀（ChIP）为基础的检测，允许病毒和细胞的蛋白质研究的影响cccDNA的迷你染色体的表观遗传修饰，染色质动力学，及转录活性]，我们建立了HBV复制的调节，在一个单元格的复制系统和在肝脏中的HBV慢性感染的病人，由cccDNA的绑定H3/H4组蛋白的乙酰化状态。我们还表明，α-干扰素（IFNα）利于长期招聘核cccDNA的微型染色体III类HDAC hSirt1的和的PRC2压制性络合物，包括转录抑制HBV在体外和体内的转录和复制共同抑制HDAC1和EZH2。
目的：我们寻求的可行性进行评估，针对表观遗传调控cccDNA的微型染色体核小不同类别的染色质修饰酶的活性的化合物，模仿IFNα对乙肝病毒的转录和复制的镇压活动。
评估方法：衣壳相关HBV-DNA（荧光定量实时PCR），，cccDNA的（TaqMan实时PCR）和pgRNA水平（实时定量PCR特异性引物），在转染的HepG2细胞中HBV基因组全长离开未经处理或处理用）类I / II和III类组蛋白去乙酰化酶抑制剂（HDACI），B）P300和PCAF组蛋白乙酰转移酶（HAT）抑制剂C），hSirt1活性剂和d）JMJD3组蛋白去甲基化酶抑制剂。
结果：我们证实了I / II类的HDACi与力量的cccDNA的绑定组蛋白乙酰化，cccDNA的转录和HBV复制。相反，合并抑制p300和：PCAF帽子（化合物EML-264）的pgRNA转录减少明显降低，HBV复制，镜像。的hSirt1 / 2活性剂的MC2562和MC2791，尽管有不同的效率，同时抑制HBV复制与cccDNA转录。 EZH2基因活性的增强作用是通过抑制组蛋白去甲基化酶的JMJD3复合MC3119> 50％的的pgRNA转录减少。值得一提的是，抑制hSirt1 / 2（MC2344）或EZH2（MC2887）强烈诱导细胞死亡，从而阻碍病毒复制的影响的评价。
结论：总之，这些结果代表了概念证明的小分子，可诱导激活hSirt1和EZH2一个“积极的后生镇压”HBV cccDNA的微小染色体的观察IFNα。

lin12345

谢谢分享 虽然翻译后很拗口

咬牙硬挺

同意楼上的