Long-Term Inhibition of HIV transcription by targeting cellular CDK9 in vivo.

通过体内靶向细胞 CDK9 长期抑制 HIV 转录。

基本信息

批准号：
8847280
负责人：
Alonso Heredia
金额：
$ 19.19万
依托单位：
UNIVERSITY OF MARYLAND BALTIMORE
依托单位国家：
美国
项目类别：
财政年份：
2014
资助国家：
美国
起止时间：
2014-05-08 至 2017-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8847280
关键词：
Aftercare Animal Model Antiviral Agents Blood Blood Chemical Analysis CD34 gene CDK9 Protein Kinase Cells Chinese Traditional Medicine Chronic Dominant-Negative Mutation Dose Drug Kinetics Drug resistance Drug toxicity Effectiveness Gene Expression Genes Genetic Transcription Goals HIV HIV Infections HIV therapy Health Human Human Genome Immunocompetent In Vitro Inbred BALB C Mice Indinavir Integrase Inhibitors Intestines Lead Life Life Cycle Stages Lymphocyte Mus Patients Plasma Protease Inhibitor Proviruses RNA RNA Interference RNA Splicing Reporting Resistance development Reverse Transcription Safety Tenofovir Testing Therapeutic Tissues Toxic effect Transplantation Viral Virion Virus antiretroviral therapy cellular targeting cytotoxicity improved in vivo indirubin inhibitor/antagonist macrophage novel pre-clinical prevent resistant strain viral RNA viral resistance

项目摘要

DESCRIPTION: Propagation of HIV in the human host requires cell entry, reverse transcription of viral RNA, integration into the human genome, transcription of the integrated provirus, and assembly/release of new virus particles. Currently there are antiretrovirals (ARTs) against each of these viral steps, except for provirus transcription. Although combinations of existing ARTs control HIV replication in most patients, drug toxicity and drug resistance are remaining concerns, arguing for the discovery of additional ARTs with novel mechanisms of action. In particular, an inhibitor of HIV transcription might both increase potency of treatment and suppress drug-resistant strains, improving the lives of patients. Our long-term goal is to improve treatment in HIV patients by targeting cellular factors essential in the virus life cycle. Cellular cyclin-dependent kinase 9 (CDK9) is required for transcription of both cellular genes and the HIV provirus. Approaches targeting CDK9 in vitro with catalytic inhibitors, RNAi, and direct inhibition using a dominant negative form, have all suggested that inhibition of HIV transcription without toxicity might be possible. Because HIV therapy is life-long, it is critical to determine safety and antiviral effectiveness of prolonged CDK9 inhibition in chronic HIV infection. We, and others, have previously shown that Indirubin 3'-monoxime (IM), a derivative of an ingredient in Chinese traditional medicine, inhibits CDK9 and HIV expression in primary lymphocytes and macrophages without cytotoxicity. In Preliminary Studies we show that IM suppresses plasma HIV RNA in NSG mice transplanted with human lymphocytes in the absence of toxicity, providing the first evidence for HIV inhibition by a CDK9 catalytic inhibitor in vivo. IM alone suppressed HIV RNA by > 2 log10 units, a magnitude of HIV reduction similar to that achieved with the NRTI EFdA in humanized mice. We also show that IM and ARTs from the NRTI, protease and integrase inhibitor classes have favorable anti-HIV interactions in vitro, suggesting IM could be used in combination with current ARTs. The goal of this application is to assess the anti-HIV potential of CDK9 inhibitors by evaluating antiviral mechanism, drug resistance and long- term toxicity in humanized mice chronically infected with HIV. Our hypothesis is that chronic treatment with CDK9 catalytic inhibitors can safely inhibit HIV transcription in vivo. We will test this hypothesis using IM or, alternatively, two novel CDK9 inhibitors. There are two Specific Aims. Specific Aim 1: To evaluate toxicity, pharmacokinetics, and antiviral activity of CDK9 inhibition in NSG mice transplanted with human CD34+ cells (HSC-NSG mice). Specific Aim 2: To evaluate mechanism of antiviral activity and long-term control of HIV in HSC-NSG mice treated with a CDK9 inhibitor. This proposal will assess the potential of blocking HIV transcription by prolonged treatment with a CDK9 inhibitor in chronically infected mice, resembling life-long therapy of HIV in patients. Successful testing of our hypothesis could improve HIV therapy by effectively targeting virus transcription.

描述：HIV 在人类宿主中的传播需要进入细胞、病毒 RNA 逆转录、整合到人类基因组中、整合的原病毒转录以及新病毒颗粒的组装/释放。目前，除了原病毒转录之外，还有抗逆转录病毒药物（ART）针对这些病毒步骤中的每一个步骤。尽管现有抗逆转录病毒疗法的组合可以控制大多数患者的艾滋病毒复制，但药物毒性和耐药性仍然令人担忧，因此需要发现具有新作用机制的其他抗逆转录病毒疗法。特别是，HIV转录抑制剂可能既可以提高治疗效力，又可以抑制耐药菌株，从而改善患者的生活。我们的长期目标是通过针对病毒生命周期中必需的细胞因子来改善艾滋病毒患者的治疗。蜂窝网络细胞周期蛋白依赖性激酶 9 (CDK9) 是细胞基因和 HIV 原病毒转录所必需的。在体外使用催化抑制剂、RNAi 和使用显性失活形式直接抑制 CDK9 的方法都表明，在没有毒性的情况下抑制 HIV 转录是可能的。由于 HIV 治疗是终生的，因此确定长期抑制 CDK9 对慢性 HIV 感染的安全性和抗病毒有效性至关重要。我们和其他人之前已经证明，靛玉红 3'-单肟 (IM)（一种中药成分的衍生物）可抑制原代淋巴细胞和巨噬细胞中 CDK9 和 HIV 的表达，且无细胞毒性。在初步研究中，我们表明，在没有毒性的情况下，IM 可抑制移植人淋巴细胞的 NSG 小鼠血浆 HIV RNA，这为 CDK9 催化抑制剂在体内抑制 HIV 提供了第一个证据。 IM 单独抑制 HIV RNA > 2 log10 单位，其 HIV 减少幅度与 NRTI EFdA 在人源化小鼠中实现的效果相似。我们还表明，来自 NRTI、蛋白酶和整合酶抑制剂类的 IM 和 ART 在体外具有良好的抗 HIV 相互作用，这表明 IM 可以与当前的 ART 联合使用。本申请的目的是通过评估慢性感染 HIV 的人源化小鼠的抗病毒机制、耐药性和长期毒性，评估 CDK9 抑制剂的抗 HIV 潜力。我们的假设是，长期使用 CDK9 催化抑制剂治疗可以安全地抑制体内 HIV 转录。我们将使用 IM 或两种新型 CDK9 抑制剂来检验这一假设。有两个具体目标。具体目标 1：在移植人 CD34+ 细胞的 NSG 小鼠（HSC-NSG 小鼠）中评估 CDK9 抑制的毒性、药代动力学和抗病毒活性。具体目标 2：评估用 CDK9 抑制剂治疗的 HSC-NSG 小鼠的抗病毒活性和长期控制 HIV 的机制。该提案将评估通过对慢性感染小鼠长期使用 CDK9 抑制剂治疗来阻断 HIV 转录的潜力，类似于对患者进行 HIV 终身治疗。成功测试我们的假设可以通过有效靶向病毒转录来改善艾滋病毒治疗。

项目成果

期刊论文数量（1）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Targeting of CDK9 with indirubin 3'-monoxime safely and durably reduces HIV viremia in chronically infected humanized mice.

DOI：
10.1371/journal.pone.0183425
发表时间：
2017
期刊：
PloS one
影响因子：
3.7
作者：
Medina-Moreno S;Dowling TC;Zapata JC;Le NM;Sausville E;Bryant J;Redfield RR;Heredia A
通讯作者：
Heredia A