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。

• 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
• 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

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：人类宿主中HIV的传播需要细胞进入，病毒RNA的逆转录，整合到人类基因组中，综合病毒的转录以及新病毒颗粒的组装/释放。目前，除了探针转录外，还针对这些病毒步骤中的每一个都有抗逆转录病毒（艺术）。尽管现有艺术的组合控制了大多数患者的艾滋病毒复制，但药物毒性和耐药性仍在关注，主张发现具有新型作用机理的其他艺术。特别是，HIV转录的抑制剂既可能增加治疗效力，又可以抑制抗药性菌株，从而改善患者的生活。我们的长期目标是通过靶向病毒生命周期中必不可少的细胞因素来改善艾滋病毒患者的治疗。细胞 细胞周期蛋白依赖性激酶9（CDK9）是细胞基因和HIV病毒的转录所必需的。用催化抑制剂，RNAi和使用显性负面形式靶向CDK9的方法都表明，可能有可能抑制HIV转录无毒性的抑制作用。由于HIV疗法是终身的，因此确定长时间CDK9抑制在慢性HIV感染中的安全性和抗病毒有效性至关重要。我们和其他人先前已经表明，中国传统医学中成分的衍生物Indirubin 3'monoxime（IM）抑制了原发性淋巴细胞和巨噬细胞中没有细胞毒性的CDK9和HIV表达。在初步研究中，我们表明，IM抑制了在没有毒性的情况下用人淋巴细胞移植的NSG小鼠中的血浆HIV RNA，这为CDK9催化抑制剂在体内抑制HIV抑制HIV的证据。 IM仅由> 2 log10单位抑制HIV RNA，这与人源化小鼠中NRTI EFDA相似的HIV减少幅度。我们还表明，来自NRTI，蛋白酶和整合酶抑制剂类别的IM和艺术在体外具有有利的抗HIV相互作用，这表明IM可以与当前的艺术结合使用。该应用的目的是通过评估人性化的小鼠长期感染HIV的抗病毒机制，耐药性和长期毒性来评估CDK9抑制剂的抗HIV潜力。我们的假设是，用CDK9催化抑制剂进行慢性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抑制剂检验该假设。有两个具体的目标。具体目的1：评估用人CD34+细胞（HSC-NSG小鼠）移植的NSG小鼠中CDK9抑制的毒性，药代动力学和抗病毒活性。具体目的2：评估用CDK9抑制剂治疗的HSC-NSG小鼠中抗病毒活性的机理和HIV的长期控制。该提案将评估通过长期用CDK9抑制剂在长期感染的小鼠中延长艾滋病毒转录的潜力，这类似于患者的HIV寿命治疗。成功测试我们的假设可以通过有效靶向病毒转录来改善HIV疗法。

项目成果

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