雷帕霉素通过增强 HBx 泛素化抑制乙型肝炎病毒共价闭合环状

StephenW

雷帕霉素通过增强 HBx 泛素化抑制乙型肝炎病毒共价闭合环状 DNA 转录
张元 1 2 3 4 , 梁丽 5 , 程盛涛 1 , 秦一平 1 , 新河 1 , 范李 6 , 戴庆 1 , 方仁 7 8 , 余海波 1 , 刘静1 , 陈娟 1 , 任继华 1 , 张振振 2 3 4
隶属关系
隶属关系

    1
    重庆医科大学传染病分子生物学教育部重点实验室，重庆，中国。
    2
    【作者单位】： 重庆医科大学儿童医院感染科;
    3
    国家儿童健康与障碍临床研究中心，儿童发育与障碍教育部重点实验室，重庆，中国。
    4
    重庆市儿童感染与免疫重点实验室，重庆，中国。
    5
    【作者单位】： 重庆大学三峡医院消化内科;
    6
    【作者单位】： 重庆医科大学第一附属医院内分泌乳腺外科;
    7
    重庆市川渝共建传染病中西医结合诊疗重点实验室
    8
    【作者单位】： 重庆市中医院检验科;

    PMID：36033851 PMCID：PMC9403416 DOI：10.3389/fmicb.2022.850087

抽象的

乙型肝炎病毒（HBV）感染仍然是世界范围内严重的公共卫生问题。干扰素和核苷（酸）类似物等抗病毒疗法可有效控制 HBV 复制，但它们无法根除慢性乙型肝炎（CHB），因为它们无法消除细胞内共价闭合环状 DNA（cccDNA）。因此，有必要开发针对 cccDNA 的新策略。由于 cccDNA 难以清除，因此 cccDNA 的转录沉默是一种可能的有效策略。 HBx在维持cccDNA的转录活性方面起着至关重要的作用，它可以成为阻断cccDNA转录的靶点。为了筛选可能有助于抗病毒治疗的新药，HiBiT 裂解检测系统检测了 2,000 种小分子化合物抑制 HBx 的能力。我们发现临床上用于预防器官移植后急性排斥反应的大环内酯类化合物雷帕霉素可显着降低HBx蛋白表达。机理研究表明，雷帕霉素通过泛素-蛋白酶体系统促进其降解，从而降低了 HBx 蛋白的稳定性。此外，雷帕霉素抑制HBV感染细胞中的HBV RNA、HBV DNA和cccDNA转录水平。此外，HBx 缺乏消除了雷帕霉素诱导的 cccDNA 转录抑制。在重组 cccDNA 小鼠模型中也证实了类似的结果。总之，我们报告了一种新的小分子雷帕霉素，它靶向 HBx 以阻断 HBV cccDNA 转录并抑制 HBV 复制。这种方法可以确定治愈 CHB 的新策略。

关键词：HBx；共价闭合环状 DNA；乙型肝炎病毒;雷帕霉素;转录。

版权所有 © 2022 Zhang, Li, Cheng, Qin, He, Li, Wu, Ren, Yu, Liu, Chen, Ren and Zhang。

StephenW

Rapamycin inhibits hepatitis B virus covalently closed circular DNA transcription by enhancing the ubiquitination of HBx
Yuan Zhang  1   2   3   4 , Liang Li  5 , Sheng-Tao Cheng  1 , Yi-Ping Qin  1 , Xin He  1 , Fan Li  6 , Dai-Qing Wu  1 , Fang Ren  7   8 , Hai-Bo Yu  1 , Jing Liu  1 , Juan Chen  1 , Ji-Hua Ren  1 , Zhen-Zhen Zhang  2   3   4
Affiliations
Affiliations

    1
    The Key Laboratory of Molecular Biology of Infectious Diseases Designated by the Chinese Ministry of Education, Chongqing Medical University, Chongqing, China.
    2
    Department of Infectious Disease, Children's Hospital of Chongqing Medical University, Chongqing, China.
    3
    National Clinical Research Center for Child Health and Disorders, Ministry of Education Key Laboratory of Child Development and Disorders, Chongqing, China.
    4
    Chongqing Key Laboratory of Child Infection and Immunity, Chongqing, China.
    5
    Department of Gastroenterology, Chongqing University Three Gorges Hospital, Chongqing, China.
    6
    Department of Endocrine and Breast Surgery, The First Affiliated Hospital of Chongqing Medical University, Chongqing, China.
    7
    Chongqing Key Laboratory of Sichuan-Chongqing Co-construction for Diagnosis and Treatment of Infectious Diseases Integrated Traditional Chinese and Western Medicine, Chongqing, China.
    8
    Department of Laboratory Medicine, Chongqing Hospital of Traditional Chinese Medicine, Chongqing, China.

    PMID: 36033851 PMCID: PMC9403416 DOI: 10.3389/fmicb.2022.850087

Abstract

Hepatitis B virus (HBV) infection is still a serious public health problem worldwide. Antiviral therapies such as interferon and nucleos(t)ide analogs efficiently control HBV replication, but they cannot eradicate chronic hepatitis B (CHB) because of their incapacity to eliminate endocellular covalently closed circular DNA (cccDNA). Thus, there is a necessity to develop new strategies for targeting cccDNA. As cccDNA is difficult to clear, transcriptional silencing of cccDNA is a possible effective strategy. HBx plays a vitally important role in maintaining the transcriptional activity of cccDNA and it could be a target for blocking the transcription of cccDNA. To screen new drugs that may contribute to antiviral therapy, the ability of 2,000 small-molecule compounds to inhibit HBx was examined by the HiBiT lytic detection system. We found that the macrolide compound rapamycin, which is clinically used to prevent acute rejection after organ transplantation, could significantly reduce HBx protein expression. Mechanistic studies demonstrated that rapamycin decreased the stability of the HBx protein by promoting its degradation via the ubiquitin-proteasome system. Moreover, rapamycin inhibited HBV RNA, HBV DNA, and cccDNA transcription levels in HBV-infected cells. In addition, HBx deficiency abrogated the inhibition of cccDNA transcription induced by rapamycin. Similar results were also confirmed in a recombinant cccDNA mouse model. In summary, we report a new small-molecule, rapamycin, which targets HBx to block HBV cccDNA transcription and inhibit HBV replication. This approach can identify new strategies to cure CHB.

Keywords: HBx; covalently closed circular DNA; hepatitis B virus; rapamycin; transcription.

Copyright © 2022 Zhang, Li, Cheng, Qin, He, Li, Wu, Ren, Yu, Liu, Chen, Ren and Zhang.

StephenW

https://www.frontiersin.org/arti ... icb.2022.850087/pdf

tim889

没啥价值 直接抑制所有transcript的ASO/RNAi 都干不掉cccDNA

即便in vitro的效果也不是很好（figure1），更别说rapamycin本身副作用就挺大，调控host的太多基因了。mTOR一搞牵一发动全身

StephenW

雷帕霉素
维基百科，自由的百科全书


雷帕霉素（Rapamune），又名西罗莫司（Sirolimus），为大环内酯类化合物，现主要运用于肾移植的抗排异治疗。其通过抑制白细胞介素-2从而阻碍激活T细胞及B细胞来抑制免疫。其首次发现于复活节岛土壤样品内吸水链霉菌(Streptomyces hygroscopicus)产物中[1]，雷帕霉素即得名于复活节岛在波利尼西亚语中的名字雷帕岛（Rapa Nui）[2][3]。雷帕霉素最初设想开发为抗真菌药物，但发现其具有强烈的免疫抑制及抗增殖作用而被放弃用于抗真菌治疗。1999年9月，美国食品药品管理局批准将雷帕霉素作为免疫抑制剂用于肾移植。2009年，美国国家老年研究所资助的研究显示雷帕霉素可延长小鼠的寿限，特别是其最大寿限可延长达60%。同时其也正被研究用于癌症等疾病的治疗。[4]

StephenW

雷帕霉素长寿：意见文章
Mikhail V. Blagosklonny 通讯作者1
作者信息 文章注释 版权和许可信息 免责声明
本文已被 PMC 中的其他文章引用。
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抽象的

从文明的曙光开始，人类就梦想着不朽。那么，为什么 mTOR 抑制剂的抗衰老特性的发现没有永远改变世界呢？我将讨论几个原因，包括对雷帕霉素、依维莫司和其他临床批准药物的实际和虚构副作用的恐惧，并认为没有真正的副作用阻止它们在今天用作抗衰老药物。此外，雷帕霉素/依维莫司的可逆（和可避免）副作用的替代方案是衰老的不可逆（和不可避免）影响：癌症、中风、梗塞、失明和过早死亡。我还将讨论为什么不使用抗衰老药物比使用抗衰老药物更危险，以及基于雷帕霉素的药物组合如何已被实施以延长人类的潜在寿命。如果您从头到尾阅读本文，您可能会意识到现在是时候了。
关键词：雷帕霉素，rapalogs，二甲双胍，衰老，抗衰老，禁食，寿命，健康寿命

“如果你等到准备好，那几乎肯定为时已晚。”赛斯·戈丁

在一种短寿命突变小鼠品系中，mTOR 抑制剂雷帕霉素（在临床上称为西罗莫司）将最大寿命延长了近三倍 [1]。尽管不那么引人注目，但雷帕霉素还可以延长正常小鼠以及酵母、蠕虫和苍蝇的寿命，并防止啮齿动物、狗、非人类灵长类动物和人类与年龄相关的疾病。雷帕霉素及其类似物依维莫司已获得 FDA 批准供人类使用，并且已安全使用了数十年。 2006年，有人提出雷帕霉素可以立即用于减缓人类衰老和所有与年龄相关的疾病[2]，成为“当今的抗衰老药物”[3]。
Rapamycin for longevity: opinion article
Mikhail V. Blagosklonnycorresponding author1
Author information Article notes Copyright and License information Disclaimer
This article has been cited by other articles in PMC.
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Abstract

From the dawn of civilization, humanity has dreamed of immortality. So why didn’t the discovery of the anti-aging properties of mTOR inhibitors change the world forever? I will discuss several reasons, including fear of the actual and fictional side effects of rapamycin, everolimus and other clinically-approved drugs, arguing that no real side effects preclude their use as anti-aging drugs today. Furthermore, the alternative to the reversible (and avoidable) side effects of rapamycin/everolimus are the irreversible (and inevitable) effects of aging: cancer, stroke, infarction, blindness and premature death. I will also discuss why it is more dangerous not to use anti-aging drugs than to use them and how rapamycin-based drug combinations have already been implemented for potential life extension in humans. If you read this article from the very beginning to its end, you may realize that the time is now.
Keywords: rapamycin, rapalogs, metformin, aging, anti-aging, fasting, lifespan, health span

“If you wait until you are ready, it is almost certainly too late.” Seth Godin

In one short-lived mutant strain of mice, the mTOR inhibitor rapamycin (known in the clinic as Sirolimus) extends maximum life span nearly three-fold [1]. Albeit less spectacularly, rapamycin also prolongs life in normal mice as well as in yeast, worms and flies, and it prevents age-related conditions in rodents, dogs, nonhuman primates and humans. Rapamycin and its analog, everolimus, are FDA approved for human use and have been used safely for decades. In 2006, it was suggested that rapamycin could be used immediately to slow down aging and all age-related diseases in humans [2], becoming an “anti-aging drug today” [3].
https://doi.org/10.18632/aging.102355

tim889

做aging的人都知道rapamycin长期副作用大，因此一部分人在搞rapamycin的alternative。FDA没有批准rapamycin用于治疗aging或aging相关的疾病。用来治疗癌症和免疫排斥也是没办法。

用于抗乙肝病毒根本不现实。即使抑制了Hbx又能怎么样，RNAi也搞不掉cccDNA

StephenW

"总之，我们报告了一种新的小分子雷帕霉素，它靶向 HBx 以阻断 HBV cccDNA 转录并抑制 HBV 复制。这种方法可以确定治愈 CHB 的新策略。"

tim889

StephenW 发表于 2022-8-30 12:54
"总之，我们报告了一种新的小分子雷帕霉素，它靶向 HBx 以阻断 HBV cccDNA 转录并抑制 HBV 复制。这种方法 ...

我想起我以前有个学生跟他本科时候的ex导师灌水，套路永远都是找了个phenotype很弱的gene，倒腾完一堆生化之后，总得在论文结尾不痛不痒加上一句“总之，我们报告了一种基因A，它和基因B互相作用。提示基因A可以成为潜在的靶点。”

lancas

这就是专业牛人对新闻的看法，我这种问外汉就看了个寂寞