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

基本信息

批准号：
8331492
负责人：
RONALD P HART
金额：
$ 19.38万
依托单位：
RUTGERS, THE STATE UNIV OF N.J.
依托单位国家：
美国
项目类别：
财政年份：
2011
资助国家：
美国
起止时间：
2011-09-15 至 2014-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8331492
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

项目摘要

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.

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。