Intranasal Nanodelivery of Oxytocin to Treat Morphine Addiction in HIV Patients by Gene Editing

通过基因编辑鼻内纳米递送催产素治疗 HIV 患者吗啡成瘾

• 批准号: 9411292
• 负责人: Rahul Dev Jayant
• 金额: $ 14.65万
• 依托单位: FLORIDA INTERNATIONAL UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-07-01 至 2019-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9411292
• 关键词: Abbreviations; Acute; Address; Aerosols; Affect; Applications Grants; Arginine; Attenuated; Base Pairing; Biological Availability; Blood - brain barrier anatomy; Brain; Cardiovascular Diseases; Cations; Cell Nucleus; Cellular Membrane; Chronic; Clinical Research; Clinical Trials; Clustered Regularly Interspaced Short Palindromic Repeats; Cocaine; Cognitive; Complex; DNA; Data; Development; Disease; Dose; Drug Addiction; Drug Delivery Systems; Drug Tolerance; Drug abuse; Endocrine; Environment; Event; Formulation; Future; Gene Delivery; Genes; Goals; Grant; Guide RNA; HIV; HIV Infections; HIV-1; HIV-associated neurocognitive disorder; Half-Life; Health; Highly Active Antiretroviral Therapy; Hippocampus (Brain); Hormones; Human; Hypothalamic structure; Impairment; In Vitro; Industrialization; Infection; Inflammation Mediators; Ingestion; Injection of therapeutic agent; Lead; Learning; Lipids; Literature; Longevity; Lung; Malignant Neoplasms; Medical; Mendelian disorder; Metabolic; Methods; Microinjections; Molecular; Morphine; Morphine Abuse; Morphine Dependence; Mus; Nature; Nebulizer; Neurologic; Neuromodulator; Neurons; Neuropeptides; Neurotransmitters; Oligopeptides; Opiates; Opioid Receptor; Oral; Oxytocin; Patients; Penetrance; Peptides; Pharmaceutical Preparations; Phase; Physical Dependence; Physiological; Plasma; Plasmids; Play; Polyethylene Glycols; Polymers; Posterior Pituitary Gland; Prevalence; Process; Production; Proteins; RNA; Reproducibility; Risk; Role; Route; Severe Combined Immunodeficiency; Signal Pathway; Signal Transduction; Site; Source; Structure; Symptoms; System; Techniques; Therapeutic; Transfection; Translating; Treatment Efficacy; Ultrasonics; Up-Regulation; Ventral Striatum; Viral Proteins; Virus; Withdrawal; addiction; basal forebrain; cytotoxicity; drug of abuse; efficacy evaluation; experimental study; gene therapy; gene translocation; human disease; hydrophilicity; hypothalamic-pituitary-thyroid axis; immune function; improved; in vivo; interest; mouse model; nanoformulation; neural circuit; neurobehavioral; neurotoxicity; neurotransmission; new technology; non-viral gene delivery; novel; pre-clinical; protein expression; relating to nervous system; response; shear stress; systemic toxicity; tat Protein; therapeutic gene; vector

Intranasal Nanodelivery of Oxytocin to Treat Morphine Addiction in HIV Patients by Gene Editing Despite significant developments, treating drug addiction in HIV-1 infected subjects remain as a challenge. Morphine has been shown to exaggerate HIV-induced risk in patients, which deteriorates the brain function and leads to dysregulation of endocrine-metabolic system. This dysregulation might lead to disturbances in the hypothalamic-pituitary-thyroid (HPT) axis, which may indirectly effect the production of Oxytocin (OXT) (neurohypophyseal nona-neuropeptide synthesized in the brain released at the posterior pituitary). Despite the extensive literature on OXT's role in addiction therapy, there are no direct studies available investigating the effects of or on OXT during concurrent Morphine addiction and HIV-1 infection. Also, it has previously been shown that exogenous OXT delivery inhibits the development of acute and chronic morphine tolerance and attenuate the various symptoms of morphine withdrawal in dose dependent manner. OXT may act as a neuromodulator on dopaminergic neurotransmission in limbic-basal forebrain structures to regulate adaptive CNS processes leading to drug addiction. Repeated morphine ingestion will upregulate mu (µ)-opioid receptor and lead to inhibition of OXT production, which may eventually cause the development of tolerance and physical dependence. Though the OXT has significant therapeutic advantages there are insufficient levels of OXT in compromised diseased state and exogenous OXT expressing source becomes an important need. Exogenous OXT delivery becomes complicated pertaining to factors like plasma half-life and poor oral bioavailability, and limited penetrance of the blood-brain barrier (BBB) due to their large size and hydrophilic nature. Therefore, delivery of a stable and efficacious active gene encoding protein directly into the brain that would reverse the effects of morphine addiction may serve as an effective approach against HIV-1 infected drug abusing subjects. Considering these preclinical limitations related to its delivery and to address this issue, we have developed a novel polyplex nanoformulation of OXT encoding CRISPR activation plasmid by lab developed nontoxic derivative of PEI [P(SiDAAr)5P3] using novel simultaneous spray (SS) technique. Our preliminary results show that SS prepared pGFP polyplex using P(SiDAAr)5P3 (non-viral transfecting agent) could form uniform, small, stable, non-toxic polyplex and achieved significant higher transfection efficiency compared to commercially available jetPEI. To translate this novel technology as anti-addiction therapeutics in HIV-1 treatment paradigm and to achieve the goals, we propose to study the effect of spraying parameters, important formulation aspects and efficacy evaluation of SS-prepared OXT- formulation against Morphine ± HIV-1 Tat challenge using primary human neurons (Aim-1). generate i nhalable aerosol formulation of OXT- P(SiDAAr)5P3 polyplex Further as a proof of concept, we propose to P(SiDAAr)5P3 polyplex and evaluate intranasal CNS delivery in OXTtm1Wsy/OXTtm1Wsy (OXT deficient- OXTDef) mice exposed to Morphine ± HIV-1 Tat (Aim-2). Data generated using this grant will be used for future R01 grant proposal.

通过基因编辑，催产素的鼻内纳米递送以治疗HIV患者的吗啡成瘾 尽管有重大的发展，但治疗HIV-1感染受试者中的药物成瘾仍然是挑战。 吗啡已显示出夸大患者的HIV引起的风险，这决定了大脑功能 并导致内分泌代谢系统失调。这种失调可能会导致灾难 下丘脑 - 垂体 - 甲状腺（HPT）轴，可能间接影响催产素（OXT）的产生 （在垂体后释放的大脑中合成的神经型非A非尿素非A-神经肽）。尽管 关于OXT在成瘾疗法中作用的广泛文献，没有可用的直接研究调查 同时吗啡成瘾和HIV-1感染过程中牛或对牛的影响。另外，它以前有 结果表明，外源OXT递送抑制了急性和慢性吗啡耐受性的发展 并以剂量依赖性方式减轻吗啡戒断的各种症状。 oxt可以充当 在边缘 - 基形前脑结构中的多巴胺能神经传递上的神经调节剂，以调节适应性 中枢神经系统过程导致吸毒成瘾。 重复的吗啡摄入将上调MU（µ） - 阿片类药物 受体并导致抑制OXT的产生，这有时可能导致耐受性的发展 和身体依赖。尽管OXT具有明显的治疗优势 在被妥协的否定状态和外源性oxt表达来源中的oxt水平成为重要 需要。外源牛的递送变得复杂与等离子体半衰期和口服差的因素有关 生物利用度和血脑屏障（BBB）的渗透率有限，因为它们的尺寸较大和亲水性 自然。因此，将稳定有效的活性基因递送，将蛋白质编码直接传递到大脑中 会扭转吗啡成瘾的影响可能是对HIV-1感染的有效方法 滥用毒品。考虑到与其交付有关的这些临床前限制，并解决此问题 问题，我们已经开发了一种新型的oxt的多种流血纳米制剂，编码CRISPR激活质粒 实验室使用新型同时喷雾（SS）技术开发了PEI [P（SIDAAR）53]的无毒衍生物。我们的 初步结果表明，SS使用p（sidaar）53（非病毒转染剂）制备了PGFP多链体 可能形成均匀，小，稳定，无毒的多体并实现明显更高的转染效率 与市售的Jetpei相比。将这种新颖的技术翻译为反瘾治疗 在HIV-1治疗范式中，为了实现目标，我们建议 研究喷涂参数的效果， 重要的公式方面和效率评估，对SS准备的OXT- 使用原代人神经元（AIM-1）抗吗啡±HIV-1 TAT挑战的公式。 生成oxt-的吸入气溶胶公式 p（sidaar）53 息肉 进一步作为证明 概念，我们建议P（Sidaar）53 息肉 和 评估Oxttm1wsy/Oxttm1Wsy（OXT缺陷型牛）小鼠中的鼻内CNS递送 ±HIV-1 TAT（AIM-2）。使用此赠款生成的数据将用于未来的R01赠款建议。