Targeting Inflammasome with stable endocannabinoid ligand AMG315. CRISPR/Cas9 and nanotechnology study in the context of HIV and cannabinoid

使用稳定的内源性大麻素配体 AMG315 靶向炎症体。

基本信息

批准号：
10085922
负责人：
Alexandros Makriyannis
金额：
$ 38.32万
依托单位：
FLORIDA INTERNATIONAL UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-07-01 至 2025-05-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10085922
关键词：
Affinity Analgesics Animal Model Anti-Inflammatory Agents Attenuated Behavioral Binding Biological Models Blood - brain barrier anatomy Brain CNR1 gene CRISPR-nanoparticles CRISPR/Cas technology Cannabinoids Cells Charge Clustered Regularly Interspaced Short Palindromic Repeats DNA Development Disease Doxycycline Drug Delivery Systems Drug Targeting Electromagnetics Electrostatics Encapsulated Endocannabinoids Genes Genetic Transcription Genome Grant Guide RNA HIV HIV Genome HIV Infections HIV Seropositivity HIV-1 Impairment In Vitro Infection Inflammasome Inflammation Legal patent Ligands Liposomes Mediating Medical Medical Marijuana Metabolic Methods Microglia Motor Activity Mus Nanotechnology Nerve Degeneration Neurocognitive Neurocognitive Deficit Neurologic Neuronal Plasticity Neurons Patients Peripheral Pharmaceutical Preparations Plants Prevention Production Property Proteins Receptor Activation Reporting Role Surface System Technology Therapeutic Time Transgenic Mice Treatment Efficacy Viral Load result analog anandamide antiretroviral therapy astrogliosis base clinical application controlled release drug of abuse endogenous cannabinoid system improved in vivo latent infection magnetic field marijuana use mouse model multidisciplinary nanocarrier nanoformulation nanoparticle nanoparticle delivery neuroAIDS neurobehavioral neurogenesis neuroinflammation neurotoxicity novel prevent research study response synthetic cannabinoid tat Genes tat Protein

项目摘要

Targeting Inflammasome with stable endocannabinoid ligand AMG315: CRISPR/Cas9 and nanotechnology study in the context of HIV and cannabinoid. PROJECT SUMMARY (ABSTRACT): Although the use of cannabis for medical purposes has shown great promise for the treatment of certain medical conditions, cannabinoid abuse exerts significant impairments in neurocognitive and behavioral functions and these effects are exacerbated in patients with HIV infection. Studies suggest that even after HIV-1 suppressing combined antiretroviral therapy (cART), HIV-1 Tat is being produced in the brain from proviral DNA and implicated as a causative agent for latent infection and development of inflammasome mediated neuroinflammation in HIV infected patients. CRISPR/Cas9 gene-editing technology has been shown by us and others to be effective for excising the HIV genome integrated into the host genome. Our preliminary studies using a recently discovered and potent metabolically stable endocannabinoid analog AMG315 demonstrate that this synthetic cannabinoid exerts anti-inflammatory properties by suppressing NLRP3 inflammasome and HIV infection. Accordingly, we hypothesize that elimination of the HIV-1 Tat gene in CNS cells using Tat specific CRISPR/Cas9 and suppression of inflammasome with CB1-specific stable endocannabinoid analog AMG315 can eliminate active HIV infection/induce permanent latency and prevent neurodegeneration, respectively. However, AMG-315 and CRISPR are impenetrable to the brain in sufficient quantities necessary to prevent HIV-infection, inflammasome activation, and subsequent neurodegeneration. To overcome this, we will use our patented magneto-electric nanoparticles (MENP) technology and liposomes to deliver CRISPR/Cas9 and AMG315, and for on-demand controlled-release. For the sustained release and to protect CRISPR from lysosomal degradation, MENP-bound CRISPR and AMG315 will be encapsulated in liposomes. Accordingly, in Specific Aim # 1, we will develop, characterize, and evaluate the delivery of Tat-specific CRISPR Cas9/gRNA and AMG315 across the in vitro BBB using MENP-based drug delivery approach to excise HIV-1 Tat gene, and attenuate cannabinoid and Tat-induced inflammasome, respectively. In Specific Aim # 2, we will study the in vivo therapeutic efficacy of MENP nanoformulation containing CRISPR, and AMG315 using doxycycline-inducible HIV-1 Tat transgenic mice (iTat) as an HIV/neuroAIDS and cannabinoid administration animal model. In Specific Aim-3, we will validate the effects of these nanoformulations on neuronal plasticity and neurocognitive functions in vivo. Successful completion of this grant will have a translational significance in preventing HIV-1 Tat and cannabinoid-mediated neurodegeneration in HIV infected cannabinoid-abusing patients. This multidisciplinary new break-through concept targeting inflammasome in the brain with stable endocannabinoid ligand AMG315 and Tat specific CRISPR/Cas9 using MENP-based technology is in response to RFA-DA-20-026 and will be useful for the suppression of HIV replication, NLRP3 inflammasome activity and to treat cannabinoid-induced neuronal impairments in HIV infected cannabinoid abusers.

使用稳定的内源性大麻素配体 AMG315 靶向炎症体：CRISPR/Cas9 和 HIV 和大麻素背景下的纳米技术研究。项目概要（摘要）：尽管大麻用于医疗目的已显示出治疗某些疾病的巨大希望医疗条件下，大麻素滥用会对神经认知和行为造成严重损害功能，并且这些影响在 HIV 感染患者中会加剧。研究表明，即使在 HIV-1 抑制联合抗逆转录病毒疗法 (cART)，HIV-1 Tat 是在大脑中产生的前病毒 DNA 并被认为是潜伏感染和炎症小体发展的病原体介导 HIV 感染患者的神经炎症。 CRISPR/Cas9基因编辑技术我们和其他人证明它可以有效切除整合到宿主基因组中的 HIV 基因组。我们的初步研究使用最近发现的有效代谢稳定的内源性大麻素类似物 AMG315 证明这种合成大麻素通过抑制 NLRP3 炎性体和 HIV 感染。因此，我们假设 HIV-1 Tat 基因的消除在中枢神经系统细胞中使用 Tat 特异性 CRISPR/Cas9 并通过 CB1 特异性稳定抑制炎症小体内源性大麻素类似物 AMG315 可以消除活跃的 HIV 感染/诱导永久潜伏并预防分别是神经退行性疾病。然而，AMG-315 和 CRISPR 无法充分穿透大脑。预防艾滋病毒感染、炎症小体激活和随后的神经退行性疾病所需的数量。为了克服这个问题，我们将使用我们的专利磁电纳米颗粒 (MENP) 技术脂质体用于递送 CRISPR/Cas9 和 AMG315，并用于按需控释。为了持续释放并保护 CRISPR 免受溶酶体降解，MENP 结合的 CRISPR 和 AMG315 将被封装在脂质体中。因此，在具体目标#1中，我们将开发、描述和评估使用基于 MENP 的药物将 Tat 特异性 CRISPR Cas9/gRNA 和 AMG315 传递到体外 BBB 切除 HIV-1 Tat 基因并减弱大麻素和 Tat 诱导的炎症体的递送方法，分别。在具体目标#2中，我们将研究 MENP 纳米制剂的体内治疗功效含有 CRISPR 和 AMG315，使用多西环素诱导的 HIV-1 Tat 转基因小鼠 (iTat) 作为 HIV/神经艾滋病和大麻素给药动物模型。在Specific Aim-3中，我们将验证效果这些纳米制剂对体内神经元可塑性和神经认知功能的影响。顺利完成这笔赠款对于预防 HIV-1 Tat 和大麻素介导的药物具有转化意义 HIV感染的大麻素滥用患者的神经退行性变。这一多学科新突破使用稳定的内源性大麻素配体 AMG315 和 Tat 特异性靶向大脑炎症体的概念使用基于 MENP 的技术的 CRISPR/Cas9 是对 RFA-DA-20-026 的回应，将有助于抑制 HIV 复制、NLRP3 炎性体活性并治疗大麻素诱导的神经元感染艾滋病毒的大麻素滥用者的损伤。