A cutaneous gene therapy for cocaine abuse and alcohol co-abuse

针对可卡因滥用和酒精滥用的皮肤基因疗法

基本信息

批准号：
10217078
负责人：
Xiaoyang Wu
金额：
$ 43.74万
依托单位：
UNIVERSITY OF CHICAGO
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-09-30 至 2024-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10217078
关键词：
Acetylcholine Agonist Alcohol abuse Alcohol consumption Alcohols Attenuated Autologous Transplantation BCHE gene Behavior Behavioral Brain Diseases Butyrylcholinesterase CRISPR/Cas technology Cells Clinical Cocaine Cocaine Abuse Cocaine Users Counseling Cues Cutaneous Development Dose Doxycycline Drug Addiction Drug abuse Eating Engineered skin Engineering Enzymes Ethanol Exhibits Exposure to FDA approved Female GLP-I receptor Gene Delivery Genes Goals Hepatocyte Human Human Engineering Hydrolysis Immune response Immunocompetent Intravenous Methods Morbidity - disease rate Mus Mutagenesis Non-Insulin-Dependent Diabetes Mellitus Nude Mice Organ Patients Peripheral Pharmaceutical Preparations Physiological Plasma Pre-Clinical Model Property Relapse Research Residual state Risk Rodent Self Administration Site Skin Skin Transplantation Skin graft Substrate Specificity Sudden Death Surface System Testing Therapeutic Therapeutic Agents Time Tissue Engineering Toxic effect Training Transplantation Treatment Protocols Work alcohol relapse alcohol use disorder behavioral response blood glucose regulation burn therapy burn wound cell type cocaethylene conditioned place preference cost drug of abuse epidermal stem cell experience gene therapy glucagon-like peptide 1 help-seeking behavior in vivo individual variation innovation male mortality novel parenteral administration prevent preventable death response skin wound stem cells therapeutic gene

项目摘要

PROJECT SUMMARY Cocaine and alcohol are commonly abused and frequently co-abused drugs. Available medications do not meet the needs for treating ongoing cocaine and alcohol abuse, relapse and co-abuse. The modified human butyrylcholinesterase (hBChE) exhibits great catalytic potency and substrate specificity for cocaine hydrolysis and is effective in reducing the behavioral and toxic effects of cocaine in rodents. The glucagon-like peptide 1 (GLP1) receptor agonists can attenuate the reinforcing properties of cocaine and alcohol in rodents. Both hBChE and GLP1 have very short half-lives in vivo, however, limiting their potential in treating cocaine abuse and co-abuse with alcohol. Cultured epidermal autografts have been clinically used for treatment of massive skin wounds for decades. Transplantation of skin grafts derived from engineered skin progenitor cells provides a novel and ideal approach for long-term and efficient delivery of therapeutic agents in vivo. We have made key technical advancement in developing a novel mouse-to-mouse skin transplantation method that allows the stable introduction of engineered epidermal progenitor cells into immunocompetent host mice. We have also used the CRISPR technology to target either an hBChE gene or a doxycycline (dox)-inducible modified GLP1 gene (DImGLP1) into epidermal progenitor cells. We then transplanted the engineered hBChE or DImGLP1 cells into mice either separately (GhBChE and GDImGLP1) or together (GBChEGLP1). GhBChE and GDImGLP1 mice did not develop cocaine- and ethanol-induced conditioned place preference (CPP) respectively. Grafting hBChE cells right after CPP expression attenuated cocaine-induced reinstatement of CPP. GBChEGLP1 mice did not develop CPP and they exhibited a lower lethality rate induced by cocaine and ethanol co-administration than those in control mice. The goal of this proposal is to take advantage of these novel platforms to test an innovative cutaneous gene therapy for cocaine abuse and co-abuse with alcohol in mice. We will also evaluate duration of hBChE and mGLP1 protection against cocaine and/or alcohol abuse, potential immune responses and approaches to reduce them. We expect the proposed cutaneous gene therapy to be long-lasting, highly specific and efficient with little individual variation. To be able to engineer stable in vivo bio-delivery systems for therapeutic genes via epidermal progenitor cells is significant because it holds potential for reducing ongoing cocaine abuse and co-abuse with alcohol and relapse in users and addicts. The proposed work will have a high impact in that results will lay key groundwork for the development of a highly personalized, long-lasting and affordable approach for combating cocaine abuse and co-abuse with alcohol.

项目摘要可卡因和酒精通常被滥用，并且经常被共同缓解。可用的药物没有满足治疗正在进行的可卡因和酗酒，复发和共同虐待的需求。经过修改的人丁甘氨酸酯酶（HBCHE）表现出可卡因水解的催化效力和底物特异性并有效地减少可卡因在啮齿动物中的行为和毒性作用。胰高血糖素样肽1 （GLP1）受体激动剂可以衰减啮齿动物中可卡因和酒精的增强特性。两个都 HBCHE和GLP1在体内的半衰期很短，限制了它们在治疗可卡因滥用方面的潜力并与酒精共滥用。培养的表皮自体移植已在临床上用于大规模治疗皮肤伤口数十年。源自工程皮肤祖细胞的皮肤移植物的移植提供一种新颖的理想方法，用于长期有效地在体内递送治疗剂。我们做了开发一种新型鼠标到小鼠皮肤移植方法的关键技术进步，该方法允许稳定地将工程表皮祖细胞引入免疫能力的宿主小鼠。我们也有使用CRISPR技术来靶向HBCHE基因或强力霉素（DOX）诱导的修改GLP1 基因（DIMGLP1）进入表皮祖细胞。然后，我们移植了工程的HBCHE或DIMGLP1 细胞分别进入小鼠（GHBCHE和GDIMGLP1）或一起（GBCHEGLP1）。 ghbche和 GDIMGLP1小鼠没有发展可卡因和乙醇引起的条件位置偏好（CPP）分别。 CPP表达后，将可卡因诱导的恢复原位嫁接HBCHE细胞 CPP。 GBCHEGLP1小鼠没有发育CPP，它们表现出可卡因和可卡因诱导的致死率较低乙醇共同给药比对照小鼠的乙醇共同给药。该提议的目的是利用这些用于测试可卡因滥用可卡因并与酒精共同滥用的创新性皮肤基因疗法的新型平台老鼠。我们还将评估HBCHE和MGLP1防止可卡因和/或酗酒的持续时间，潜在的免疫反应和减少它们的方法。我们期望提出的皮肤基因治疗是持久的，高度特异性和有效的，几乎没有个体变异。能够设计通过表皮祖细胞的治疗基因的体内生物传递系统稳定是显着的因为它具有减少正在进行的可卡因滥用并与酒精共同滥用并复发的潜力在用户和吸毒者中。拟议的工作将产生很大的影响，因为结果将是关键开发高度个性化，持久和负担得起的方法的基础打击可卡因滥用并与酒精共同滥用。