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：
10197872
负责人：
Alexandros Makriyannis
金额：
$ 37.11万
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/10197872
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.

用稳定的内源性大麻素配体AMG315靶向炎症体：CRISPR/CAS9和纳米技术研究在HIV和大麻素的背景下。项目摘要（摘要）：尽管将大麻用于医疗目的已显示出对某些治疗的巨大希望医疗状况，大麻素滥用在神经认知和行为方面造成了重大损害 HIV感染患者的功能和这些作用加剧了。研究表明，即使之后 HIV-1抑制组合抗逆转录病毒疗法（CART），HIV-1 TAT正在大脑中产生前病毒DNA，并被视为潜在感染和发育的病因 HIV感染患者的介导的神经炎症。 CRISPR/CAS9基因编辑技术已经由我们和其他人表明，可以有效地切除整合到宿主基因组中的HIV基因组。我们的使用A的初步研究最近发现且有效的代谢稳定的内源性大麻素类似物 AMG315证明该合成大麻素通过抑制施加抗炎特性 NLRP3炎性体和HIV感染。因此，我们假设消除HIV-1 TAT基因在中枢神经系统细胞中，使用特异性CRISPR/CAS9和CB1特异性稳定的炎症体抑制内源性大麻素模拟AMG315可以消除活跃的HIV感染/诱导永久潜伏期并防止神经变性分别。但是，AMG-315和CRISPR在足够的大脑中无法接受防止HIV感染，炎症体激活和随后的神经变性所需的数量。为了克服这一点，我们将使用我们的专利磁电纳米颗粒（MENP）技术和脂质体提供CRISPR/CAS9和AMG315，以及按需控制释放的脂质体。为了持续为了保护CRISPR免受溶酶体降解的释放，Menp-Bound CRISPR和AMG315将是封装在脂质体中。因此，在特定的目标＃1中，我们将开发，表征和评估使用基于MENP的药物，在体外BBB上递送TAT特异性CRISPR CAS9/GRNA和AMG315 消费税HIV-1 TAT基因的输送方法，并减弱大麻素和TAT诱导的炎性体，分别。在特定的目标＃2中，我们将研究MENP纳米成型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）作为艾滋病毒/神经辅助和大麻素给药动物模型。在特定的AIM-3中，我们将验证效果这些对体内神经元可塑性和神经认知功能的纳米形成。成功完成这笔赠款的转化意义将具有预防HIV-1 TAT和大麻素介导的 HIV感染大麻素虐待患者的神经变性。这个多学科的新分手具有稳定的内源性大麻素配体AMG315和特异性的概念靶向大脑中的炎症体使用基于Menp的技术的CRISPR/CAS9响应RFA-DA-20-026，对抑制HIV复制，NLRP3炎性体活动并治疗大麻素诱导的神经元艾滋病毒感染大麻素施用者的损害。