Phenotypic analysis of human iPSC carrying addiction-associated gene variants

携带成瘾相关基因变异的人类 iPSC 表型分析

• 批准号: 8250138
• 负责人: RONALD P HART
• 金额: $ 23.1万
• 依托单位: RUTGERS, THE STATE UNIV OF N.J.
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-09-15 至 2013-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8250138
• 关键词: Addictive Behavior; Adoption; Adult; Affect; Alleles; Amino Acids; Archives; Authorization documentation; Behavior; Biochemical; Biological Assay; Blood; Blood specimen; Brain Diseases; Calcium; Cell Culture Techniques; Cell Line; Cell Surface Receptors; Cell model; Cell physiology; Cells; Child Abuse; Cholinergic Receptors; Chronic; Collection; Crime; Cultured Cells; DNA; Data; Drug Addiction; Drug abuse; Electrophysiology (science); Employment; Episome; Family; Gene Expression; Genes; Genetic Variation; Genomics; Genotype; Goals; Health; Housing; Human; Image; Individual; Inherited; Intervention; Laboratories; Lymphocyte; Maps; Methods; Modeling; Molecular; Molecular Models; Morphine; Mutate; Mutation; National Institute of Drug Abuse; National Institute of Mental Health; National Institute on Alcohol Abuse and Alcoholism; Neuronal Differentiation; Neurons; Nicotine; Nicotinic Receptors; Opioid; Opioid Receptor; Patients; Pharmaceutical Preparations; Pharmacology; Phenotype; Physiological; Physiology; Property; Protocols documentation; Publishing; Reporting; Research; Research Personnel; Resources; Risk Factors; Sampling; Screening procedure; Second Messenger Systems; Signal Transduction; Societies; Source; Staging; Stem Cell Research; Stem cells; Study models; Substance abuse problem; Substance of Abuse; System; Techniques; Technology; Testing; Time; Universities; Variant; Viral; addiction; base; cell bank; cell type; cost; embryonic stem cell; expectation; gene function; genetic variant; immunocytochemistry; induced pluripotent stem cell; molecular marker; molecular modeling; nerve stem cell; novel; novel strategies; pluripotency; release of sequestered calcium ion into cytoplasm; repository; response; second messenger; vector

DESCRIPTION (provided by applicant): Identification of genetic variations associated with addictive behaviors provides a novel opportunity for using cell cultures to model molecular and cellular mechanisms that underlie addiction. The observed genetic variations have been mapped to amino acid changes in cell surface receptors, presumably affecting neuronal circuits involved in addiction behaviors. However, cellular context is likely to be important in determining the function of these genes. Construction of induced pluripotent stem cells (iPSC) from adult cells derived from drug abusing individuals carrying known genetic variants provides a means for developing physiologically relevant culture systems for understanding addiction. For the culture cell model to be useful it must not only express the affected gene but it must also provide an appropriate cellular context for studying pharmacology or cell signaling. There is no clear expectation about what type of iPSC-derived cultures would be useful in studying the cellular physiology altered by gene variants. Our hypothesis is that the altered physiology of genetic variants associated with addiction liability can be modeled in cultured neurons derived from iPSCs constructed from donor lymphocyte samples. We propose to construct several iPSC lines from donor lymphocyte samples, differentiate these iPSC into functional neurons, and then to develop methods to assay possible phenotypic differences between variant-derived cells and wild-type. These cells will be valuable for identifying cell and molecular responses to substances of abuse, to examine the effects of a known genotype on the cellular phenotype, as well as to develop novel approaches for pharmacologic intervention. PUBLIC HEALTH RELEVANCE: Drug abuse and addiction are a major burden to society, with the total cost of substance abuse in the U.S. exceeding $500 billion annually. Addiction is a chronic brain disease that leads to a compulsive desire for drugs despite significantly harmful consequences, including direct effects on health as well as detriments to society, including disintegration of families, child abuse, loss of employment, and a broad array of crimes. While addiction may be treatable, the discovery that some forms of addiction have an inherited component demonstrates that physiology contributes to addictive behaviors. To exploit new techniques of cellular reprogramming to prepare human neuronal cultures, we will establish induced pluripotent cell (iPSC) lines derived from donor blood samples carrying mutations in two genes known to be risk factors for addiction-an acetylcholine receptor and an opioid receptor. The blood lymphocyte samples already exist in the NIDA Center for Genomic Studies collection and will be used with permission from the collecting investigator. These iPSC will then be induced to differentiate into specific types of neurons for use in investigating the molecular bases of addiction and for screening new treatment therapies.

描述（由申请人提供）：与成瘾行为相关的遗传变异的识别为使用细胞培养物来建模成瘾基础的分子和细胞机制提供了新的机会。观察到的遗传变异已映射到细胞表面受体的氨基酸变化，可能会影响成瘾行为涉及的神经元回路。但是，细胞环境对于确定这些基因的功能可能很重要。从滥用携带已知遗传变异的个体的成年细胞中诱导多能干细胞（IPSC）的诱导多能干细胞（IPSC），为开发生理相关的培养系统提供了一种理解成瘾的方法。为了使培养细胞模型有用，它不仅必须表达受影响的基因，而且还必须为研究药理学或细胞信号传导提供适当的细胞环境。关于哪种类型的IPSC衍生培养物对于研究基因变异改变的细胞生理学将是有用的。我们的假设是，与成瘾责任相关的遗传变异的生理变化可以在源自供体淋巴细胞样品构建的IPSC的培养神经元中进行建模。我们建议构建与供体淋巴细胞样品的几种IPSC系，将这些IPSC区分为功能性神经元，然后开发方法来分析变异衍生的细胞和野生型之间可能的表型差异。这些细胞对于鉴定细胞和分子对滥用物质的反应将是有价值的，以检查已知基因型对细胞表型的影响，并开发出新颖的药理干预方法。 公共卫生相关性：药物滥用和成瘾是社会的重大负担，美国每年滥用药物的总成本超过5000亿美元。成瘾是一种慢性脑疾病，尽管有明显的有害后果，但仍会导致对药物的强迫性渴望，包括对健康的直接影响以及对社会的损害，包括对家庭的瓦解，虐待儿童，就业丧失，就业和广泛的犯罪。尽管成瘾可能是可以治疗的，但某些形式的成瘾具有遗传成分的发现表明生理学有助于成瘾行为。为了利用细胞重编程的新技术来制备人神经元培养物，我们将建立诱导的多能细胞（IPSC）线，这些细胞（IPSC）系来自供体血液样本，这些供体血液样品中携带突变的两个基因，已知是成瘾的危险因素 - 乙酰胆碱受体和一个阿片类药物。血液淋巴细胞样品已经存在于NIDA基因组研究中心收集中心，并将在收集研究者的许可下使用。然后，这些IPSC将被诱导分化为特定类型的神经元，以研究成瘾的分子碱基并筛选新的治疗疗法。