Dissecting the mechanism of action of CHD1L, a novel regulator of HIV-1 infection

剖析 HIV-1 感染的新型调节因子 CHD1L 的作用机制

• 批准号: MR/S009752/1
• 负责人: Andrew Lever
• 金额: $ 96.03万
• 依托单位: University of Cambridge
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2019
• 资助国家: 英国
• 起止时间: 2019 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=MR%2FS009752%2F1
• 关键词: Dissecting; mechanism; action; CHD1L; novel

HIV, the virus that causes AIDS, remains an important cause of ill health and mortality, with 37 million people living with the virus worldwide and annually being responsible for around 1 million deaths. This is despite significant progress in antiviral treatments. The virus interacts with infected human cells, and some of these interactions involve host defence molecules that critically affect the ability of the virus to complete its life cycle. This knowledge has informed the targeting of crucial viral processes in current treatment regimes. Most screening studies on how human genetics affect HIV replication have been conducted on European populations, despite more than 70 % of infected individuals and nearly half of new infections each year being in Africa. A recent study conducted by our collaboration has revealed a novel gene which has an enormous impact on HIV replication in patients of African ancestry. Upon infection, the level of the HIV in the body of the infected patient reaches a steady state and this level is a key factor in determining how quickly the immune system is compromised by the virus. This, in turn, is an important determinant of how long an HIV-infected individual takes to progress to AIDS. Our genomic study shows that a higher level of expression of a gene, chd1l, is associated with a lower steady state level of virus. The strength of the association between the level of protein produced by this gene CHD1L and the HIV level is equivalent to that of well described mutations in CCR5, one of the molecules necessary for most commonly transmitted viruses to gain entry into a cell, and these mutations are known to confer significant protection from HIV in patients harbouring them. Thus CHD1L has the potential to have an enormous impact on HIV replication.It remains unclear, however, how CHD1L is able to inhibit HIV infection. Here we propose research to understand the interaction between CHD1L and HIV. We aim to pinpoint the step(s) in the virus life cycle at which CHD1L may be exerting an effect, through quantifying the markers of the different stages in cells with and without CHD1L. CHD1L unwinds the packaging of DNA in response to specific signals in the cell. There are two candidate parts of the life cycle where this could be important. 1. In order to express its proteins, HIV inserts its genome into the host DNA in a process called integration. CHD1L could affect the efficiency and/or specificity of this process. 2. DNA also needs to be unwound for the encoded proteins to be expressed; again CHD1L could affect this process. We will also determine whether CHD1L binds to any specific HIV proteins by using highly sensitive techniques to see what CHD1L is bound to in infected and uninfected cells. In this way, we will identify the proteins and pathways in the cell through which CHD1L affects HIV replication. We will also identify features of the CHD1L protein that are essential for its ability to limit HIV infection by making changes to key parts of the protein and monitoring the ability of modified versions of the protein to inhibit HIV infection. Interactions with specific viral or cellular proteins could be exploited therapeutically to generate new treatments for HIV. The knowledge generated by our proposed experiments is necessary to translate our initial observation into clinically useful treatment for this devastating virus. The group of experts assembled to carry out this work are uniquely well-placed position to do this due to their expertise in the technology required for this work and their links to clinical and commercial partners.

HIV 是导致艾滋病的病毒，仍然是导致健康不良和死亡的一个重要原因，全世界有 3700 万人感染这种病毒，每年导致约 100 万人死亡。尽管抗病毒治疗取得了重大进展，但情况仍然如此。该病毒与受感染的人类细胞相互作用，其中一些相互作用涉及宿主防御分子，这些分子严重影响病毒完成其生命周期的能力。这些知识为当前治疗方案中关键病毒过程的靶向提供了信息。大多数关于人类遗传学如何影响艾滋病毒复制的筛查研究都是在欧洲人群中进行的，尽管每年超过 70% 的感染者和近一半的新感染者都在非洲。我们合作最近进行的一项研究揭示了一种新基因，它对非洲血统患者的艾滋病毒复制具有巨大影响。感染后，感染患者体内的艾滋病毒水平达到稳定状态，这一水平是决定免疫系统被病毒损害的速度的关键因素。反过来，这又是艾滋病毒感染者发展为艾滋病所需时间的重要决定因素。我们的基因组研究表明，基因 chd1l 的较高表达水平与较低的病毒稳态水平相关。该基因 CHD1L 产生的蛋白质水平与 HIV 水平之间的关联强度相当于 CCR5 中已描述的突变的强度，CCR5 是最常见传播的病毒进入细胞所必需的分子之一，并且已知这些突变可以为携带它们的患者提供显着的保护，使其免受 HIV 感染。因此，CHD1L有可能对HIV复制产生巨大影响。然而，目前尚不清楚CHD1L如何能够抑制HIV感染。在这里，我们提出研究来了解 CHD1L 和 HIV 之间的相互作用。我们的目标是通过量化有和没有 CHD1L 的细胞中不同阶段的标记来查明 CHD1L 可能发挥作用的病毒生命周期中的步骤。 CHD1L 响应细胞中的特定信号而解开 DNA 包装。在生命周期的两个候选部分中，这一点可能很重要。 1. 为了表达其蛋白质，HIV 将其基因组插入宿主 DNA，这一过程称为整合。 CHD1L 可能会影响该过程的效率和/或特异性。 2. DNA也需要解绕才能表达编码的蛋白质； CHD1L 可能再次影响这一过程。我们还将使用高度敏感的技术来确定 CHD1L 是否与任何特定的 HIV 蛋白结合，以了解 CHD1L 在感染和未感染细胞中的结合情况。通过这种方式，我们将确定细胞中 CHD1L 影响 HIV 复制的蛋白质和途径。我们还将通过改变蛋白质的关键部分并监测该蛋白质的修饰版本抑制 HIV 感染的能力，来确定 CHD1L 蛋白质的特征，这些特征对于其限制 HIV 感染的能力至关重要。与特定病毒或细胞蛋白的相互作用可以在治疗上用于产生针对艾滋病毒的新疗法。我们提出的实验产生的知识对于将我们最初的观察转化为针对这种毁灭性病毒的临床有用治疗是必要的。由于他们在这项工作所需的技术方面的专业知识以及与临床和商业合作伙伴的联系，为开展这项工作而聚集的专家组处于独特的有利地位。

项目成果

Analysis of SAMHD1 Restriction by Flow Cytometry in Human Myeloid U937 Cells.

通过流式细胞术分析人髓样 U937 细胞中的 SAMHD1 限制。

• DOI: 10.17863/cam.66961
• 发表时间: 2021
• 作者: Ordonez P

A novel, sensitive dual-indicator cell line for detection and quantification of inducible, replication-competent latent HIV-1 from reservoir cells

• DOI: 10.1038/s41598-019-55596-8
• 发表时间: 2019-12-18
• 期刊: SCIENTIFIC REPORTS
• 影响因子: 4.6
• 作者: Salasc, Fanny;Gludish, David W.;Mok, Hoi-Ping
• 通讯作者: Mok, Hoi-Ping

Analysis of SAMHD1 Restriction by Flow Cytometry in Human Myeloid U937 Cells

• DOI: 10.3791/62502
• 发表时间: 2021-06-01
• 期刊: JOVE-JOURNAL OF VISUALIZED EXPERIMENTS
• 影响因子: 1.2
• 作者: Ordonez，Paula;Bishop，Kate N.;Groom，Harriet Cordelia Theed
• 通讯作者: Groom，Harriet Cordelia Theed

A novel, sensitive dual-indicator cell line for detection and quantification of inducible, replication-competent latent HIV-1 from reservoir cells.

一种新型、灵敏的双指示细胞系，用于检测和定量来自储存细胞的可诱导的、具有复制能力的潜伏 HIV-1。

• DOI: 10.17863/cam.46209
• 发表时间: 2019
• 作者: Salasc F