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.

项目概要可卡因和酒精是常被滥用且经常同时滥用的药物。现有药物不满足治疗持续可卡因和酒精滥用、复发和共同滥用的需求。改造后的人类丁酰胆碱酯酶 (hBChE) 对可卡因水解表现出强大的催化效力和底物特异性并能有效减少可卡因对啮齿动物的行为和毒性影响。胰高血糖素样肽1 (GLP1) 受体激动剂可以减弱可卡因和酒精对啮齿动物的增强作用。两个都然而，hBChE 和 GLP1 的体内半衰期非常短，限制了它们治疗可卡因滥用的潜力并与酒精同时滥用。培养的自体表皮移植物已在临床上用于治疗大面积皮肤伤口长达数十年。来自工程化皮肤祖细胞的皮肤移植物的移植提供了一种在体内长期有效递送治疗剂的新颖且理想的方法。我们已经做了开发新型小鼠至小鼠皮肤移植方法的关键技术进步使将工程表皮祖细胞稳定引入免疫活性宿主小鼠。我们还有使用 CRISPR 技术靶向 hBChE 基因或多西环素 (dox) 诱导的修饰 GLP1 基因（DImGLP1）进入表皮祖细胞。然后我们移植了工程化的 hBChE 或 DImGLP1 将细胞分别（GhBChE 和 GDImGLP1）或一起（GBChEGLP1）注入小鼠体内。 GhBChE 和 GDImGLP1 小鼠没有出现可卡因和乙醇诱导的条件性位置偏好 (CPP) 分别。 CPP 表达后立即移植 hBChE 细胞可减弱可卡因诱导的恢复菲律宾人民党。 GBChEGLP1 小鼠没有发生 CPP，并且它们表现出由可卡因和与对照小鼠相比，乙醇共同给药的效果更好。该提案的目标是利用这些用于测试可卡因滥用和与酒精同时滥用的创新皮肤基因疗法的新平台老鼠。我们还将评估 hBChE 和 mGLP1 针对可卡因和/或酒精滥用的保护持续时间，潜在的免疫反应以及减少这些反应的方法。我们期望所提出的皮肤基因治疗持久、高度特异性和有效，个体差异很小。能够进行工程设计通过表皮祖细胞提供治疗基因的稳定体内生物递送系统具有重要意义因为它有可能减少持续的可卡因滥用以及与酒精的共同滥用和复发在用户和成瘾者中。拟议的工作将产生重大影响，其结果将成为关键为开发高度个性化、持久且负担得起的方法奠定基础打击可卡因滥用以及与酒精的共同滥用。