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.

成瘾可以被定义为尽管有不良后果但对物质使用失去控制。成瘾 法律的和非法的物质破坏了成瘾者及其家人的生活， 给社会带来巨大的负担和代价。基于分子和细胞的机制有助于 成瘾的开始和发展仍有待阐明。据估计，40 - 60%的 成瘾的脆弱性可能归因于遗传变异。多个染色体区域已经被 与成瘾有关的包括那些含有多巴胺转运蛋白（DAT）和囊泡单胺 VMAT2基因。目前的努力，以了解如何多态性在这些单胺 导致成瘾分子机制的转运蛋白由于不能 直接询问患者的神经细胞类型。患者特异性细胞来源携带特异性 能够稳健且可重复地分化成特定神经谱系的遗传变异体不 存在.我们建议开发一种基于细胞的系统，通过这种系统， 功能分析以询问成瘾的分子机制。为了实现这一目标，我们 已经制定了一个尖端的建议，其中包括多个学科的技能和专业知识。在 目的1我们将从成瘾中获得并鉴定患者特异性的诱导多能干细胞 携带单胺转运体多态性的患者和对照。由于中脑多巴胺能系统 在天然和药物相关的奖赏途径中发挥重要作用，并代表了 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鉴定它们的分子特征（即A9， A10，中脑边缘或中皮层多巴胺能神经元）。在目标3中，我们将描述、比较和 从控制和成瘾中对这些患者特异性iPS细胞衍生的多巴胺能神经元进行功能分析 hDAT 1和hVMAT 2基因多态性患者。有很大的潜力，患者特异性 iPS细胞技术通过提供 基因上不同的功能性人类细胞来源。通过完成本提案中提出的目标， 我希望能提供一个详细的特点，多巴胺能神经传递功能的患者， 并提供深入了解这种复杂疾病的病理生理学以及 单胺转运蛋白基因变异与成瘾的关系。我们建立了一个跨学科的 该团队结合了对吸毒者、神经分化和多巴胺能神经系统的人种学研究的优势， 功能分析，以及多能性和iPS细胞，以询问有关细胞和 导致成瘾的分子功能障碍我们希望我们的研究结果 与理解和治疗这种人类疾病直接相关。