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

鼻内纳米递送催产素通过基因编辑治疗HIV患者的吗啡成瘾 尽管取得了重大进展，但治疗HIV-1感染受试者的药物成瘾仍然是一个挑战。 吗啡已被证明会夸大患者中艾滋病毒诱导的风险，从而使大脑功能恶化 并导致内分泌代谢系统失调。这种失调可能会导致 下丘脑-垂体-甲状腺（HPT）轴，可能间接影响催产素（OXT）的产生 （神经垂体九肽-脑内合成的神经肽，在垂体后叶释放）。尽管 关于OXT在成瘾治疗中的作用的大量文献中，没有直接的研究可以调查 吗啡成瘾和HIV-1感染同时发生时OXT的影响。此外，此前 已经显示外源性OXT递送抑制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可以作为 边缘-基底前脑结构多巴胺能神经传递调节适应性 导致药物成瘾的CNS过程。 重复摄入吗啡将上调μ-阿片类药物 受体，并导致抑制OXT的生产，这可能最终导致耐受性的发展 身体依赖。尽管OXT具有显著的治疗优势，但仍存在不足。 受损疾病状态中的OXT水平和外源性OXT表达源成为重要的 需要的外源性OXT递送变得复杂，涉及血浆半衰期和口服不良等因素。 由于它们的大尺寸和亲水性， 自然因此，将编码蛋白质的稳定和有效的活性基因直接递送到脑中， 将逆转吗啡成瘾的影响可能成为对抗HIV-1感染的有效方法 药物滥用主题。考虑到这些与其输送相关的临床前限制， 问题，我们已经开发了编码CRISPR活化质粒的OXT的新型多聚复合物纳米制剂， 实验室使用新型同时喷雾（SS）技术开发了PEI [P（SiDAAr）5P3]的无毒衍生物。我们 初步结果表明，SS使用P（SiDAAr）5P3（非病毒分离剂）制备pGFP多聚物， 能形成均匀、细小、稳定、无毒的复合物，转染效率显著提高 与市售的jetPEI相比。将这种新技术转化为抗成瘾疗法 在HIV-1治疗范例中，为了实现目标，我们建议 研究了喷雾参数对喷雾效果的影响， SS制备的OXT的重要制剂方面和功效评价- 使用原代人神经元（Aim-1），对吗啡± HIV-1达特攻击的制剂进行了研究。 产生OXT的可吸入气雾剂制剂， P（SiDAAr）5P3 polyplex 进一步证明 提出了P（SiDAAr）5P3的概念 polyplex 和 评价暴露于吗啡的OXTtm1Wsy/OXTtm1Wsy（OXT缺陷-OXTDef）小鼠中的鼻内CNS递送 ± HIV-1达特（Aim-2）。使用此补助金生成的数据将用于未来的R01补助金提案。

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.