HIV-1 Integrase

HIV-1整合酶

基本信息

批准号：
10702556
负责人：
stephen h hughes
金额：
$ 137.38万
依托单位：
DIVISION OF BASIC SCIENCES - NCI
依托单位国家：
美国
项目类别：
财政年份：
资助国家：
美国
起止时间：
至
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10702556
关键词：
Active Sites Acute Animal Model Animals Anti-HIV Agents Anti-Retroviral Agents Antigens Antiviral Agents Antiviral Therapy Behavior Biological Assay Blood Cancer Etiology Cell Fraction Cells Chemistry Clonal Expansion Collaborations Complex Cultured Cells DNA DNA Integration Data Development Drug Kinetics Drug resistance Enzymes FDA approved Genes Genome Goals Growth HIV HIV Infections HIV-1 HIV-1 integrase HIV/AIDS Human Human Herpesvirus 4 Human Herpesvirus 8 Human Papillomavirus Immunologic Deficiency Syndromes In Vitro Individual Infection Integrase Integrase Inhibitors Integration Host Factors Introns Laboratories Life Cycle Stages Macaca Malignant Neoplasms Modification Mutation Nucleotides Oncogene LCK Oncogenes Oncogenic Viruses Patients Peripheral Blood Mononuclear Cell Persons Pharmaceutical Preparations Play Process Proliferating Proteins Provirus Integration Proviruses Reaction Research Resistance Resolution Retroviridae Risk Role STAT3 gene Site Structure T-Cell Development T-Cell Lymphoma T-Lymphocyte Testing Time Toxic effect United States National Institutes of Health Variant Viral Virion Virus Work antiretroviral therapy cancer risk cell transformation cytokine experimental study in vivo inhibitor integration site mutant nanomolar prevent resistant strain side effect structural biology treatment strategy viral DNA virology

项目摘要

INSTI development: The development of new, broadly effective anti-HIV drugs that have little or no toxicity is a high-priority NIH goal for HIV/AIDS research. Because integration is an essential step in the virus life cycle, IN is an important target for antiretroviral drugs. The HIV-1 integration reaction proceeds in two steps, both of which are carried out by IN. In the first step (3' processing, or 3'P), IN removes two nucleotides from the 3' ends of the linear viral DNA. In the second step, IN inserts the trimmed 3' ends of the viral DNA into host DNA; this reaction is called strand transfer, or ST. IN has only one active site which carries out both the 3'P and ST reactions. The four approved anti-IN drugs (raltegravir, elvitegratir, dolutegravir, and bictegravir) all target the ST reaction, and are, for this reason, called integrase strand transfer inhibitors, or INSTIs. Despite their relatively recent development, INSTIs are potent drugs with few side effects that are becoming increasingly important in antiretroviral therapies (ART). Cabotegravir has been formulated with rilpivirine and this combination has shown promise in long-term treatment strategies. However, as is the case with all anti-HIV drugs, INSTIs select for resistant strains of HIV. We are developing new INSTIs that are broadly effective against the known drug-resistant mutants. We have made excellent progress in developing IN inhibitors that have low nanomolar potency against the WT virus in a one-round inhibition assay, retain potency against a broad panel of resistant mutants, and show little or no toxicity in cultured cells. Our collaborator, Dmitry Lyumkis, has solved high resolution structures of HIV-1 IN (both WT and drug resistant) in complexes with both FDA approved INSTIs and with the most promising of the compounds developed and synthesized in the laboratory of another collaborator, Dr. Terry Burke. Although we continue to make and test new INSTIs, we have made sufficient progress that, with help from the NCI, pharmacokinetic testing is being done in macaques on the best of our compounds. Integration site analysis: There are millions of retroviral integration sites in the host cell genome, but for many retroviruses, including HIV, integration is far from random. We have been studying the distribution of HIV proviruses (integration sites) in both cultured cells and patients. HIV integration is known to favor the bodies of highly expressed genes. In work that was done in cultured cells, we were part of a collaboration that showed that the host protein HRP2 could replace LEDGF in directing HIV-1 integration to the bodies of highly expressed genes. We also showed that the host factor CPSF6 plays a key role in guiding the preintegration complex to regions of the genome that are gene rich and contain numerous highly expressed genes. We and others have used integration site analysis to show that there is extensive clonal expansion of HIV-infected cells in patients on ART. We also showed, in HIV-infected individuals on long-term ART, that more than 40% of the infected T cells are in clones and that, in some cases, proviruses integrated into six oncogenes can contribute to clonal expansion of infected T cells. Some of these highly expanded clones carry infectious proviruses and a small fraction of the cells in the clones can release infectious virions into the blood. Thus, expanded clones of HIV infected T cells that carry intact infectious proviruses represent a large fraction of the infected cells that comprise the reservoir that has made it impossible to cure HIV infections with the available anti-viral therapies. The reservoir, which arises early in infection, persists on ART and can rekindle an active infection if ART is discontinued. Because the number of possible sites of HIV DNA integration is very large, each infected cell, and all of its descendants, can be identified by the site where the provirus is integrated. Surprisingly, we found that the distribution of HIV proviruses, even after years of complete virological suppression, in which the majority of the infected surviving cells are removed from the originally infected cells by many divisions, closely resembles the distribution of the proviruses at the time the cells were initially infected. In our experiments, the initial distribution was represented by the distribution of the proviruses in PBMC acutely infected in vitro. In addition to a modest modification to the initial distribution by a positive selection for proviruses integrated in six known oncogenes, the distribution of proviruses is also modified by selection against cells with proviruses integrated in highly expressed genes. Thus, although proviruses in oncogenes can cause the extensive in a small fraction of the clones of infected cells, our data imply that most important factors causing clonal expansion of HIV infected T cells are antigen and/or cytokine stimulation. Our data also strongly support the idea that the integration site of a provirus has little if any effect on the expression of the integrated provirus. Retroviruses can cause cancers in animals by integrating in or near oncogenes. Although HIV-1 infection increases the risk of cancer, most of the risk is associated with HIV induced immunodeficiency which allows oncogenic viruses to proliferate (EBV, KSHV, HPV). As was described in the previous paragraph, HIV-1 proviruses integrated in six oncogenes can cause clonal expansion of infected T cells in vivo; however, the infected cells are not transformed. Until very recently, it was generally believed that HIV-1 does not cause cancer directly. However, we recently showed that HIV-1 proviruses integrated in the first introns of STAT3 and LCK can play an important role in the development of T cell lymphomas. The development of these cancers appears to be a multi-step process involving additional non-viral mutations, which could help explain why T cell lymphomas are rare in people living with HIV-1 infections.

Insti Development：几乎没有毒性或没有毒性的新的，广泛有效的抗HIV药物是艾滋病毒/艾滋病研究的高优先性NIH目标。由于整合是病毒生命周期中必不可少的一步，因此IN是抗逆转录病毒药物的重要靶标。 HIV-1整合反应分为两个步骤，两步都由IN进行。在第一步（3'加工或3'p）中，从线性病毒DNA的3'端去除两个核苷酸。在第二步中，在将病毒DNA的修剪成3'末端插入到宿主DNA中；该反应称为链转移或st。在只有一个活跃位点，可以同时执行3'p和ST反应。四种批准的抗内部药物（Raltegravir，Elvitegratir，Dolutegravir和Bictegravir）均针对ST反应，因此，称为Integrase Strand转移抑制剂或Instis。尽管它们的发展相对较新，但研究所是有效的药物，几乎没有副作用，在抗逆转录病毒疗法（ART）中变得越来越重要。 Cabotegravir已配制了Rilpivirine，这种组合在长期治疗策略中表现出了希望。但是，与所有抗HIV药物一样，研究所选择HIV的抗性菌株。我们正在开发针对已知的耐药突变体的新研究，这些研究物广泛有效。我们在单轮抑制测定中对WT病毒的抑制剂的发展方面取得了出色的进步，保留对宽阔的抗性突变体的效力，并且在培养细胞中几乎没有毒性。我们的合作者DMITRY LYUMKIS已通过FDA批准的Instis，以及在另一个合作者Terry Burke博士的实验室中开发和合成的化合物中最有前途的化合物，解决了HIV-1的高分辨率结构（WT和抗药性）。尽管我们继续制作和测试新的研究所，但我们取得了足够的进步，在NCI的帮助下，在猕猴中正在以最好的化合物进行药代动力学测试。整合位点分析：宿主细胞基因组中有数百万个逆转录病毒整合位点，但是对于包括HIV在内的许多逆转录病毒，整合远非随机。我们一直在研究培养细胞和患者中HIV病毒（整合位点）的分布。众所周知，HIV融合有利于高度表达基因的身体。在培养细胞中完成的工作中，我们是一项合作的一部分，该协作表明宿主蛋白HRP2可以替代LEDGF将HIV-1整合引导到高度表达基因的体内。我们还表明，宿主因子CPSF6在指导富基因富基因组并包含许多高度表达基因的基因组区域的前整合复合物中起关键作用。我们和其他人使用整合位点分析表明，ART患者的HIV感染细胞有广泛的克隆扩张。我们还显示，在感染了长期ART的HIV感染者中，超过40％的感染T细胞在克隆中，在某些情况下，在某些情况下，将预科病毒纳入了六种癌基因，可以导致感染T细胞的克隆扩张。这些高度扩展的克隆中有一些具有感染性病毒，而克隆中的一小部分细胞可以将传染性病毒体释放到血液中。因此，携带完整感染性病毒的HIV感染T细胞的扩展克隆代表了构成储层的大部分感染细胞，这使得无法通过可用的抗病毒疗法治愈HIV感染。在感染早期出现的水库一直存在于艺术上，如果停产，则可以重新激发主动感染。由于HIV DNA整合的可能位点的数量非常大，因此每个受感染的细胞及其所有后代都可以通过整合病毒的位点来识别。令人惊讶的是，我们发现，即使经过多年的完全病毒学抑制，HIV病毒的分布，其中大多数被感染的幸存细胞被许多分裂从最初感染的细胞中移除，与最初感染细胞时的原病毒分布非常相似。在我们的实验中，初始分布由PBMC在体外急性感染的PBMC中的分布表示。除了通过对六个已知肿瘤基因的阳性选择对初始分布进行适度的修改，还通过对具有积分为高度表达基因的病毒的细胞进行选择来修改Provirus的分布。因此，尽管癌基因中的原病毒可能导致一小部分感染细胞的克隆，但我们的数据表明，引起HIV感染T细胞克隆膨胀的最重要因素是抗原和/或细胞因子刺激。我们的数据还强烈支持这样一个想法，即病毒病毒的集成站点对综合病毒的表达几乎没有任何影响。逆转录病毒会通过整合在癌基因中或附近引起动物的癌症。尽管HIV-1感染增加了癌症的风险，但大多数风险与HIV诱导的免疫缺陷有关，这使致癌病毒得以增殖（EBV，KSHV，HPV）。如前一段中所述，在六种癌基因中整合的HIV-1病毒可能会导致体内感染T细胞的克隆扩张。但是，被感染的细胞没有转化。直到最近，人们普遍认为HIV-1不会直接引起癌症。但是，我们最近表明，在STAT3和LCK的第一个内含子中整合的HIV-1 Provirus可以在T细胞淋巴瘤的发展中起重要作用。这些癌症的发展似乎是一个多步骤过程，涉及其他非病毒突变，这可以帮助解释为什么在患有HIV-1感染的人中很少使用T细胞淋巴瘤。

项目成果

期刊论文数量（30）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Development of tricyclic hydroxy-1H-pyrrolopyridine-trione containing HIV-1 integrase inhibitors.

DOI：
10.1016/j.bmcl.2011.03.047
发表时间：
2011-05-15
期刊：
Bioorganic & medicinal chemistry letters
影响因子：
2.7
作者：
Zhao XZ;Maddali K;Metifiot M;Smith SJ;Vu BC;Marchand C;Hughes SH;Pommier Y;Burke TR Jr
通讯作者：
Burke TR Jr

6,7-Dihydroxyisoindolin-1-one and 7,8-Dihydroxy-3,4-Dihydroisoquinolin- 1(2H)-one Based HIV-1 Integrase Inhibitors.

基于 6,7-二羟基异吲哚啉-1-酮和 7,8-二羟基-3,4-二氢异喹啉-1(2H)-酮的 HIV-1 整合酶抑制剂。

DOI：
10.2174/1568026615666150813150058
发表时间：
2016
期刊：
Current topics in medicinal chemistry
影响因子：
3.4
作者：
Zhao,XueZhi;Metifiot,Mathieu;Smith,StevenJ;Maddali,Kasthuraiah;Marchand,Christophe;Hughes,StephenH;Pommier,Yves;BurkeJr,TerrenceR
通讯作者：
BurkeJr,TerrenceR

Correction to: An analytical pipeline for identifying and mapping the integration sites of HIV and other retroviruses.

更正：用于识别和绘制 HIV 和其他逆转录病毒整合位点的分析管道。

DOI：
10.1186/s12864-020-06924-0
发表时间：
2020
期刊：
BMC genomics
影响因子：
4.4
作者：
Wells,DariaW;Guo,Shuang;Shao,Wei;Bale,MichaelJ;Coffin,JohnM;Hughes,StephenH;Wu,Xiaolin
通讯作者：
Wu,Xiaolin

Retrovirus Integration Database (RID): a public database for retroviral insertion sites into host genomes.

DOI：
10.1186/s12977-016-0277-6
发表时间：
2016-07-04
期刊：
Retrovirology
影响因子：
3.3
作者：
Shao W;Shan J;Kearney MF;Wu X;Maldarelli F;Mellors JW;Luke B;Coffin JM;Hughes SH
通讯作者：
Hughes SH

What Integration Sites Tell Us about HIV Persistence.