Intranasal Nanodelivery of Oxytocin to Treat Morphine Addiction in HIV Patients by Gene EditingDespite 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 functionand leads to dysregulation of endocrine-metabolic system. This dysregulation might lead to disturbances inthe 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). Despitethe extensive literature on OXT's role in addiction therapy, there are no direct studies available investigatingthe effects of or on OXT during concurrent Morphine addiction and HIV-1 infection. Also, it has previouslybeen shown that exogenous OXT delivery inhibits the development of acute and chronic morphine toleranceand attenuate the various symptoms of morphine withdrawal in dose dependent manner.OXT may act as aneuromodulator on dopaminergic neurotransmission in limbic-basal forebrain structures to regulate adaptiveCNS processes leading to drug addiction.Repeated morphine ingestion will upregulate mu (µ)-opioidreceptor and lead to inhibition of OXT production, which may eventually cause the development of toleranceand physical dependence. Though the OXT has significant therapeutic advantages there are insufficientlevels of OXT in compromised diseased state and exogenous OXT expressing source becomes an importantneed. Exogenous OXT delivery becomes complicated pertaining to factors like plasma half-life and poor oralbioavailability, and limited penetrance of the blood-brain barrier (BBB) due to their large size and hydrophilicnature. Therefore, delivery of a stable and efficacious active gene encoding protein directly into the brain thatwould reverse the effects of morphine addiction may serve as an effective approach against HIV-1 infecteddrug abusing subjects. Considering these preclinical limitations related to its delivery and to address thisissue, we have developed a novel polyplex nanoformulation of OXT encoding CRISPR activation plasmid bylab developed nontoxic derivative of PEI [P(SiDAAr)5P3] using novel simultaneous spray (SS) technique. Ourpreliminary 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 efficiencycompared to commercially available jetPEI. To translate this novel technology as anti-addiction therapeuticsin HIV-1 treatment paradigm and to achieve the goals, we propose tostudy 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)5P3polyplexFurther as a proofof concept, we propose to P(SiDAAr)5P3polyplexandevaluate 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.