Exploration of novel block-and-lock agents alone and in combination for HIV remission in humanized mice

探索新型阻断剂和联合用药在人源化小鼠中缓解 HIV

基本信息

批准号：
10714365
负责人：
J. Victor Garcia-Martinez
金额：
$ 90.32万
依托单位：
UNIVERSITY OF FLORIDA
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-03-20 至 2028-02-29
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10714365
关键词：
Acceleration Aging Aldosterone Antagonists Attention BLT mice Biological Assay Bone Marrow CD4 Positive T Lymphocytes Cancer Patient Cell Line Cells Chronic Clinic Clinical Clinical Research Clinical Trials Cultured Cells DNA Integration Defect Disease Progression Disease remission Dose Drug resistance ERCC3 gene Epigenetic Process Evolution FDA approved Future Gene Expression Genes Genetic Transcription Goals HIV HIV Infections HIV antiretroviral HIV tat Protein HIV-1 Heart failure Hypertension In Vitro Individual Infection Inflammation Integration Host Factors Interruption Length Liver Mineralocorticoid Receptor Modeling Monitor Outcome Pathway interactions Persons Pharmaceutical Preparations Positioning Attribute Provirus Integration Proviruses Residual state Resistance Rest Spironolactone Testing Thymus Gland Time Tissues Transcription Initiation Site Viral Viral Proteins Viral reservoir Viremia Virus antagonist antiretroviral therapy comorbidity cortistatin cost detection limit disease transmission epigenetic silencing experience genetic inhibitor humanized mouse immune activation improved improved outcome in vivo in vivo Model inhibitor insight interest knock-down large scale production memory CD4 T lymphocyte mouse model new therapeutic target novel promoter reactivation from latency small hairpin RNA targeted agent transcription factor transcription factor TFIIH transcriptome sequencing transcriptomics viral RNA viral rebound viral resistance

项目摘要

Abstract HIV-1 transcriptional inhibitors have immense potential in functional cure approaches and could transform the way we treat HIV infections. Unlike current antiretroviral therapy (ART), transcriptional inhibitors offer the prospect of reducing residual viremia derived from reservoir of long-lived cells containing integrated proviruses, likely reducing ongoing the chronic immune activation, inflammation and HIV-associated co-morbidities still experienced by ART-adherent individuals living with HIV. Furthermore, we believe transcriptional inhibitors are amenable to block-and-lock functional cure approaches, aimed at the durable suppression of HIV in the absence of daily therapy, through permanent epigenetic silencing of integrated proviruses. This hypothesis was founded on the activity of the potent Tat inhibitor didehydro-Cortistatin A (dCA). In in vitro and in vivo models of HIV latency, dCA inhibition of HIV transcription over time prompts the viral promoter into deep transcriptional inhibition, limiting viral reactivation upon treatment interruption or with latency reactivating agents (LRAs). Despite their great potential, there are still no HIV transcriptional inhibitors in the clinic, and challenges with the cost of large-scale production of dCA are slowing its progression towards clinical studies. Here we propose to investigate the repurposing the FDA approved aldosterone antagonist Spironolactone (SP) for HIV transcriptional inhibition. An off-target activity of SP is the degradation of the XPB subunit of the general transcription factor TFIIH, a key player in RNAPII initiation at the transcriptional start site (TSS) of genes. We demonstrated in vitro that SP treatment or shRNA knockdown of XPB selectively inhibits HIV transcription and blocks viral reactivation from latency without global transcriptomic defects. This study highlighted the host factor XPB as a novel drug target and SP as a potential block-and-lock agent. Here we propose to explore the potential of SP, alone or in combination with dCA, as a block-and-lock agent in the humanized bone-marrow, thymus liver (BLT) mouse model of HIV infection by: 1) Determine the relationship between SP treatment length with residual viremia levels in tissues and correlates of chronic immune activation/inflammation in HIV infected ART- suppressed BLT mice.; 2) Assess the ability of SP to maintain deep latency as a single drug in the absence of ART and study viral resistance evolution; 3) Impact of dCA and SP in combination as front-line therapy on the size of the established viral reservoir and time to viral rebound. We predict that longer treatment lengths of HIV infected BLT mice with SP will correlate with improved reduction of low-grade HIV persistent transcription from the viral reservoir and likely chronic immune activation. Importantly, we seek to demonstrate that once deep transcriptional suppression is established, SP alone blocks viral rebound. In addition, when used as front-line therapy we expect a reduction in the size of the established viral reservoir and the combination with dCA will improve the outcome. This study will provide an important proof-of-concept for the use of transcriptional inhibitors to treat people living with HIV and explore optimal experimental settings to be tested in future clinical trials

抽象的 HIV-1转录抑制剂在功能治疗方法中具有巨大的潜力，可以改变我们治疗艾滋病毒感染的方式。与目前的抗逆转录病毒疗法（ART）不同，转录抑制剂提供减少从含有综合病毒的长寿命细胞的储层中衍生出的残留病毒血症的前景，可能还会减少持续的慢性免疫激活，炎症和与HIV相关的合并症由艾滋病毒感染的艺术遵守者体验。此外，我们认为转录抑制剂是适合阻止锁定功能治愈方法，目的是在不存在的情况下持久抑制HIV 日常治疗，通过对综合病毒的永久表观遗传沉默。这个假设建立了关于有效TAT抑制剂Didehydro-Cortistatin A（DCA）的活性。在体外和体内HIV模型中潜伏期，DCA对HIV转录的抑制会促使病毒启动子进入深层转录抑制作用，在治疗中断或潜伏期重新激活剂（LRAS）时限制病毒重新激活。尽管具有很大的潜力，但诊所仍没有HIV转录抑制剂，并且挑战大规模生产DCA的成本正在减缓其向临床研究的发展。在这里，我们建议研究重新利用FDA批准的醛固酮拮抗剂螺内酯（SP）用于HIV转录抑制。 SP的脱靶活动是一般XPB亚基的降解转录因子TFIIH是基因转录起始位点（TSS）的RNAPII启动的关键参与者。我们在体外证明了XPB的SP处理或shRNA敲低有选择地抑制HIV转录和阻止病毒重新激活，而没有全局转录组缺陷。这项研究突出了宿主因素 XPB是一种新型的药物靶标，SP作为潜在的阻滞剂。在这里，我们建议探索潜力 SP，单独或与DCA结合，作为人源骨ro的块和锁定药（BLT）HIV感染的小鼠模型：1）确定SP处理长度与残留的关系在HIV感染的ART中，组织中的病毒血症水平和慢性免疫激活/炎症的相关性被抑制的BLT小鼠。 2）评估SP在没有的情况下将SP保持深延延迟为单一药物的能力艺术和研究病毒抗性演化； 3）DCA和SP组合作为前线疗法的影响已建立的病毒储存库的大小和病毒反弹的时间。我们预测艾滋病毒的治疗长度较长具有SP的感染BLT小鼠将与减少低级HIV持续转录的降低相关病毒储量和可能的慢性免疫激活。重要的是，我们试图证明这一次建立转录抑制作用，仅SP会阻止病毒反弹。另外，当用作前线治疗我们预计已建立的病毒储层的大小和与DCA的组合会减少改善结果。这项研究将为使用转录抑制剂提供重要的概念验证治疗艾滋病毒感染者并探索将来临床试验中测试的最佳实验环境