Impact of μ-opioid receptor (MOR) splice variant interactions with the chemokine receptor CCR5 in the context of morphine and HIV-1 entry inhibitor therapy

在吗啡和 HIV-1 进入抑制剂治疗背景下，μ-阿片受体 (MOR) 剪接变异体与趋化因子受体 CCR5 相互作用的影响

• 批准号: 9308938
• 负责人: NAZIRA EL-HAGE
• 金额: $ 18.13万
• 依托单位: FLORIDA INTERNATIONAL UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-07-01 至 2019-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9308938
• 关键词: Address; Affect; Agonist; Alpha Cell; Antiviral Agents; Applications Grants; Area; Astrocytes; Binding; Biological Assay; Biological Process; Brain; CCR5 gene; Cell Surface Receptors; Cell fusion; Cells; Development; Disease; Disease Progression; Epitopes; Fishes; Genetic Polymorphism; Goals; Grant; HIV; HIV Entry Inhibitors; HIV Receptors; HIV encephalitis; HIV-1; Heterodimerization; Human; Immunoprecipitation; Individual; Inflammation; Investigation; Knowledge; Mass Spectrum Analysis; Mediating; Mediation; Methods; Microglia; Morphine; N-terminal; Neuraxis; Neurocognitive Deficit; Neuroglia; Neurologic; Opiates; Opioid; Pathogenesis; Pharmaceutical Preparations; Plasmids; Process; Protein Isoforms; RNA Splicing; Regulation; Reporting; Role; Signal Transduction; Transfection; Treatment outcome; Variant; Viral; base; cell type; chemokine; chemokine receptor; clinically relevant; design; drug abuser; inhibitor/antagonist; mu opioid receptors; neuroAIDS; opioid abuse; opioid use; overexpression; public health relevance; receptor; synergism

 DESCRIPTION (provided by applicant): Growing evidence suggests that opioid abuse exacerbates neuroAIDS and these effects are mainly mediated through the μ-opioid receptor (MOR). There is also evidence to suggest that MOR-1 may have a direct role in HIV type-1 (HIV-1) replication and pathogenesis. MOR is not a single entity, but instead exists as multiple isoforms that differs in function. Despite significant fundamental differences, most studies examining MOR have focused on the canonical MOR-1, and this is particularly true in studies of opioid drug-HIV interactions. Although MOR-1- HIV-1interactions have been studied extensively, the role of MOR splice variant regulation by HIV and, conversely, MOR variant regulation of HIV cellular binding and entry is an unexplored area. Therefore, given that MOR-1 is thought to interact with HIV co-receptors such as CCR5 via heterodimerization and/or downstream signaling, the goal of this grant is to address the variation of MOR in relation to CCR5 mediation of HIV-associated processes in the CNS in the context of morphine and the HIV-1 entry inhibitor maraviroc. Preliminary studies found that: (i) MOR-1 may not be the predominate form of MOR on all CNS cell types, (ii) the N-terminal variant MOR-1K differs functionally in cellular signaling compared to MOR-1 (iii) although both astrocytes and microglia harbors multiple MOR variants, astrocytes harbors the most (iv) MOR-CCR5 interaction occurs in both microglia and astrocytes, although more so in astrocytes and (iv) morphine the predominant agonist of MOR was able to abolish the antiviral effect of the CCR5-mediated viral entry inhibitor maraviroc in a MOR-dependent manner in both astrocytes and microglia. The present proposal is designed to 1. (i) identify the MOR splice variants that interact with the chemokine and HIV (co-) receptor CCR5 in glia (astroglia and microglia) and (ii) determine how these interactions affect viral entry in opioid using individuals in the context of the clinically relevant viral entry inhibitor maraviroc which targets CCR5-mediated entry. The findings from this project could further define the mechanisms in the progression of HIV-related complications in opioid abusers by targeting specific MOR variants and whether a particular cell type is modulating the vulnerability to these complications in the CNS. These findings may also extend to further develop HIV entry inhibitors targeting this interaction.

 描述（由申请人提供）：越来越多的证据表明阿片类药物滥用可加重神经艾滋病，这些作用主要通过μ-阿片受体（莫尔）介导。也有证据表明，莫尔-1可能在HIV-1型（HIV-1）复制和发病机制中起直接作用。莫尔不是单一实体，而是作为功能不同的多种同种型存在。尽管存在显著的根本差异，但大多数研究莫尔的研究都集中在经典的莫尔-1上，这在阿片类药物-HIV相互作用的研究中尤其如此。尽管莫尔-1- HIV-1相互作用已被广泛研究，但HIV对莫尔剪接变异体的调节作用，以及相反，HIV对莫尔变异体结合和进入细胞的调节作用仍是一个未探索的领域。因此，考虑到莫尔-1被认为通过异源二聚化和/或下游信号传导与HIV共受体（如CCR 5）相互作用，该授权的目标是在吗啡和HIV-1进入抑制剂马拉韦罗的背景下，解决与CNS中HIV相关过程的CCR 5介导相关的莫尔变异。初步研究发现：（i）莫尔-1可能不是所有CNS细胞类型上莫尔的主要形式，（ii）与莫尔-1相比，N-末端变体莫尔-1 K在细胞信号传导方面功能不同（iii）尽管星形胶质细胞和小胶质细胞都具有多种莫尔变体，但星形胶质细胞具有最多的MOR-CCR 5相互作用（iv）MOR-CCR 5相互作用发生在小胶质细胞和星形胶质细胞中，尽管在星形胶质细胞中更是如此，和（iv）吗啡，莫尔的主要激动剂，能够在星形胶质细胞和小胶质细胞中以MOR依赖性方式消除CCR 5介导的病毒进入抑制剂马拉韦罗的抗病毒作用。本提案旨在1. (i)鉴定与胶质细胞（星形胶质细胞和小胶质细胞）中的趋化因子和HIV（共）受体CCR 5相互作用的莫尔剪接变体，和（ii）在靶向CCR 5介导的进入的临床相关病毒进入抑制剂马拉韦罗的背景下，使用个体确定这些相互作用如何影响病毒进入阿片样物质。该项目的研究结果可以通过靶向特定的莫尔变体以及特定细胞类型是否调节CNS中对这些并发症的易感性来进一步确定阿片类药物滥用者中HIV相关并发症进展的机制。这些发现也可能扩展到进一步开发针对这种相互作用的HIV进入抑制剂。