Derivation and dopaminergic differentiation of human drug addict-specific iPS cel

人吸毒者特异性 iPS 细胞的衍生和多巴胺能分化

• 批准号: 8324558
• 负责人: Yu Luo
• 金额: $ 23.55万
• 依托单位: CASE WESTERN RESERVE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-09-01 至 2014-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8324558
• 关键词: Addictive Behavior; Adult; Alcohol or Other Drugs use; Alcohols; Biological Assay; Cell Line; Cell physiology; Cells; Characteristics; Complex; DNA; DNA Fingerprinting; Derivation procedure; Development; Discipline; Disease; Drug Addiction; Drug Controls; Drug user; Ethnography; Family; Fibroblasts; Functional disorder; Gene Expression; Genes; Genetic; Genetic Polymorphism; Genomics; Goals; Health; Human; Human Development; Illicit Drugs; Individual; Karyotype determination procedure; Lead; Legal; Link; Midbrain structure; Minisatellite Repeats; Mixed Function Oxygenases; Molecular; Molecular Target; Neurons; Pathway interactions; Patients; Pharmaceutical Preparations; Play; Pluripotent Stem Cells; Population; Protocols documentation; Research; Resources; Rewards; Role; Sampling; Skin; Societies; Somatic Cell; Source; Substance abuse problem; Substance of Abuse; System; Testing; Therapeutic; Tobacco; Validation; World Health Organization; addiction; adverse outcome; base; brain cell; calbindin; cell type; cohort; cost; dopamine transporter; dopaminergic differentiation; dopaminergic neuron; drug addict; drug of abuse; genetic variant; human disease; induced pluripotent stem cell; insight; monoamine; neurotransmission; novel; pluripotency; promoter; psychostimulant; relating to nervous system; response; skills; social; stem; stem cell technology; vesicular monoamine transporter

Addiction can be defined as a loss of control of substance use despite adverse consequences. Addiction to legal and illegal substances destroys the lives of both addicted subjects and their families, exerting an enormous cost and burden on society. The molecular- and cellular-based mechanisms that contribute to the initiation and development of addiction remain to be elucidated. Estimates have suggested that 40-60% of the vulnerability to addiction may be attributable to genetic aberrations. Multiple chromosomal regions have been linked to addiction including those containing the dopamine transporter (DAT) and vesicular monoamine transporter (VMAT2) genes. Current efforts to understand how polymorphisms in these monoamine transporters contribute to the molecular mechanisms of addiction are severely hindered by the inability to directly interrogate neural cell types from the patients. Patient-specific sources of cells carrying specific genetic variants that are capable of robust and reproducible differentiation into specific neural lineages do not exist. We propose to develop a cell-based system whereby neural cells from afflicted individuals can be functionally assayed to interrogate the molecular mechanisms underlying addiction. To achieve this goal we have developed a cutting-edge proposal that that incorporates the skill and expertise of multiple disciplines. In Aim 1 we will derive and characterize patient-specific, induced pluripotent stem (iPS) cells from addiction patients and controls that carry monoamine transporter polymorphisms. Since midbrain dopaminergic system play a prominent role in natural and drug related reward pathways and represent a common substrate for drugs of abuse, in Aim 2 we will differentiate patient-specific iPS cells line into dopaminergic neurons and carry out a detailed and functional characterization of these cells to identify their molecular characteristics (i.e. A9, A10, mesolimbic or mesocortical dopaminergic neurons). In Aim 3, we will characterize, compare, and functionally assay these patient-specific, iPS cell-derived dopaminergic neurons from control and addiction patients that carry polymorphisms for hDAT1 and hVMAT2 gene. There is great potential for patient-specific iPS cell technology to profoundly impact our understanding of human development and disease by providing genetically distinct, functional sources of human cells. By completing the aims set forth in this proposal we expect to provide a detailed characterization of dopaminergic neurotransmission function in patients afflicted with addiction and provide insight into the pathophysiology of this complex disease as well as the contribution of genetic variants in monoamine transporter genes to addiction. We have established an interdisciplinary team that combines strengths in ethnographic study of drug addicts, neural differentiation and dopaminergic function analysis, as well as pluripotency and iPS cells to interrogate novel questions about the cellular and molecular dysfunction that contributes to addiction. We expect that results from our studies will have immediate relevance to the understanding and treatment of this human disease.

成瘾可以定义为尽管产生不良后果但仍对物质使用失去控制。上瘾 合法和非法物质摧毁了成瘾者及其家人的生活， 给社会带来巨大的成本和负担。基于分子和细胞的机制有助于 成瘾的发生和发展仍有待阐明。据估计，40-60% 容易上瘾可能归因于基因畸变。多个染色体区域已被 与成瘾有关的物质，包括含有多巴胺转运蛋白 (DAT) 和囊泡单胺的物质 转运蛋白 (VMAT2) 基因。目前努力了解这些单胺的多态性 转运蛋白对成瘾分子机制的贡献受到严重阻碍 直接询问患者的神经细胞类型。患者特定来源的细胞携带特定的 能够稳健且可重复地分化为特定神经谱系的遗传变异体并不 存在。我们建议开发一种基于细胞的系统，通过该系统可以将来自患病个体的神经细胞 进行功能分析以探究成瘾背后的分子机制。为了实现这个目标我们 制定了一项尖端提案，其中融合了多个学科的技能和专业知识。在 目标 1 我们将从成瘾中衍生并表征患者特异性诱导多能干 (iPS) 细胞 携带单胺转运蛋白多态性的患者和对照。由于中脑多巴胺能系统 在自然和药物相关的奖赏途径中发挥着重要作用，并代表了共同的底物 滥用药物，在目标 2 中，我们将患者特异性 iPS 细胞系分化为多巴胺能神经元，并携带 对这些细胞进行详细的功能表征，以确定其分子特征（即 A9、 A10，中脑边缘或中皮质多巴胺能神经元）。在目标 3 中，我们将描述、比较和 对这些患者特异性、iPS 细胞衍生的多巴胺能神经元进行控制和成瘾功能分析 携带 hDAT1 和 hVMAT2 基因多态性的患者。针对特定患者的治疗潜力巨大 iPS 细胞技术通过提供深刻影响我们对人类发育和疾病的理解 人类细胞的遗传独特的功能来源。通过完成本提案中规定的目标，我们 期望提供患者多巴胺能神经传递功能的详细特征 与成瘾并提供深入了解这种复杂疾病的病理生理学以及贡献 单胺转运蛋白基因的遗传变异与成瘾的关系。我们建立了一个跨学科的 该团队结合了吸毒者人种学研究、神经分化和多巴胺能方面的优势 功能分析，以及多能性和 iPS 细胞，以询问有关细胞和细胞的新问题 导致成瘾的分子功能障碍。我们期望我们的研究结果能够 与了解和治疗这种人类疾病直接相关。