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

基因编辑催产素鼻腔纳米给药治疗HIV患者吗啡成瘾 尽管取得了重大进展，但治疗HIV-1感染者的药物成瘾仍然是一个挑战。 研究表明，吗啡会加重患者感染艾滋病毒的风险，从而恶化大脑功能。 并导致内分泌代谢系统失调。这种不规范可能会导致 下丘脑-垂体-甲状腺(HPT)轴，可能间接影响催产素(OXT)的产生 (神经垂体素--在脑中合成的神经肽，在垂体后叶释放)。尽管 关于奥施他命在成瘾治疗中的作用的大量文献，没有可用的直接研究 OXT对吗啡成瘾和HIV-1感染的影响。此外，它之前也曾 研究表明，外源性OXT可抑制急性和慢性吗啡耐受的发展 并以剂量依赖方式减轻吗啡戒断的各种症状。 OXT可以充当 神经调节剂对边缘-基底前脑结构多巴胺能神经传递的调节作用 中枢神经系统的过程导致药物成瘾。 反复摄入吗啡会上调u(µ)-阿片类药物 受体并导致OXT产生抑制，最终可能导致耐受性的发展 和身体上的依赖。尽管OXT具有显著的治疗优势，但仍存在不足 疾病状态下的OXT水平和外源OXT表达来源成为重要的 需要。外源性OXT的交付变得复杂，与血浆半衰期和口腔不良等因素有关 血脑屏障(BBB)的生物利用度和有限的穿透性，这是因为它们的大小和亲水性 大自然。因此，将稳定有效的编码蛋白质的活性基因直接送入大脑 逆转吗啡成瘾的效果可能是对抗HIV-1感染的有效方法 吸毒的受试者。考虑到与其交付相关的这些临床前限制并解决这一问题 问题，我们开发了一种新的编码CRISPR激活质粒的OXT多链纳米配方，通过 Lab使用新的同步喷雾(SS)技术开发了无毒的PEI[P(SiDAAr)5P3]衍生物。我们的 初步结果表明，SS使用P(SiDAAr)5P3(非病毒转染剂)制备了pGFP多聚体 可形成均匀、小、稳定、无毒的多聚体，获得显著较高的转染率 与商业上可用的JetPEI相比。将这项新技术转化为抗成瘾疗法 在艾滋病毒-1治疗模式中，为了实现这些目标，我们建议 研究了喷雾参数的影响， SS-OXT-1的重要配方及药效评价 使用原代人神经元对抗吗啡±HIV-1 TAT攻击的配方(AIM-1)。 产生OXT-的可吸入气雾剂配方- P(SiDAAr)5P3 复式 进一步作为证据 在概念上，我们提出了P(SiDAAr)5P3 复式 和 吗啡对OXTtm1Wsy/OXTtm1Wsy(OXT缺陷-OXTDef)小鼠鼻内中枢神经系统传递的影响 ±HIV-1Tat(AIM-2)。使用此赠款生成的数据将用于未来的R01赠款提案。

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.