Cellular and genomic mechanisms of the impact of ethanol on human neural model

乙醇对人类神经模型影响的细胞和基因组机制

• 批准号: 10453317
• 负责人: ZHIPING P. PANG
• 金额: $ 56.13万
• 依托单位: RBHS-ROBERT WOOD JOHNSON MEDICAL SCHOOL
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-02-05 至 2023-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10453317
• 关键词: 3-Dimensional; ATAC-seq; Acute; Address; Affect; Architecture; Astrocytes; Behavior; Biochemistry; Biological Models; Brain; Brain Diseases; Cells; Characteristics; Chromatin; Chronic; Coculture Techniques; Code; Collection; Communities; Complex; Data; Development; Electrophysiology (science); Environmental Risk Factor; Epigenetic Process; Ethanol; Family; Foundations; Functional disorder; Funding; Gene Expression; Genes; Genetic; Genetic Risk; Genetic study; Genomics; High Prevalence; Human; Human Genetics; Image; Impairment; In Vitro; Individual; Induced pluripotent stem cell derived neurons; Inflammasome; Knowledge; Link; Measures; Mediating; Mental disorders; Microglia; Minor; Modeling; Molecular; Morbidity - disease rate; Morphology; Mus; Nature; Neuroglia; Neuroimmune; Neurons; Opioid; Organoids; Pathogenicity; Pathway interactions; Patients; Phenotype; Process; Proteins; Reproducibility; Research; Resolution; Resources; Risk; Role; Societies; Stress; Study models; Synaptic Transmission; System; Testing; Untranslated RNA; Variant; alcohol effect; alcohol exposure; alcohol use disorder; cell type; cytokine; disorder risk; functional genomics; gene network; genetic makeup; genetic risk factor; genetic variant; genome-wide analysis; genomic data; induced pluripotent stem cell; insight; live cell imaging; mortality; neural circuit; neural model; neurogenesis; neuroinflammation; neuropsychiatric disorder; novel therapeutics; polygenic risk score; relating to nervous system; response; risk variant; sex; single-cell RNA sequencing; socioeconomics; stem; synaptic function; therapeutically effective; transcriptomics

Abstract and summary Alcohol use disorders (AUD) are complex behaviors accompanied by substantial morbidity, mortality and societal expense. Both genetic and environmental factors contribute to AUD. Despite progress in the human genetics of AUD, especially the identification of genome-wide significant AUD genetic risk variants, the neural basis of AUD in humans is largely unknown. Over the past five years, we have provided compelling evidence that: 1) Human neuronal cells derived from induced pluripotent stem (iPS) cells can be used as a tractable model to study neuropsychiatric disorders including AUD; 2) Ethanol exposure results in an inflammasome response in human neurons; 3) Human neurons carrying OPRM1 A118G minor gene variants showed enhanced sensitivity to opioids and ethanol; 4) Ethanol causes gene expression changes in both human neurons and glial cells; and 5) Microglia-containing 3D neural cultures can be a powerful system to study neuroinflammatory microglia-neuronal interactions. These premises provide a foundation for further mechanistic studies of the genetic and molecular underpinnings of AUD in human iPS cell-derived neural models. We also identified key gaps of knowledge in utilizing human neurons as a model system to study AUD that need to be filled, particularly: 1) how ethanol affects neuro-glial interactions in a human neural context is not known; and 2) how ethanol affects neurogenesis in a 3D context is not known. Moreover, 3) because of the polygenic nature of AUD, the contribution of single gene to AUD risk is likely small and the phenotypical manifestation is strongly influenced by individual’s genetic makeup. To address these outstanding questions, we hypothesize that ethanol impairs neuronal function via affecting neuroglial interactions, which is influenced by individual polygenic risk backgrounds. We have selected 36 subjects of both sexes with either extremely high polygenic risk score (PRS, top 10%ile, n=18 AUD) or low PRS (bottom 10%ile, n=18, no AUD) to test this hypothesis. We will differentiate iPS cells derived from these subjects into both 2D and 3D (i.e. brain organoids) neuronal cells co-cultured with human astrocytes and human microglia. Upon exposure to ethanol, these neural cells are subjected to a combination of morphological, immunocytochemical, electrophysiological, live cell imaging and genomic analyses to unravel the mechanism(s) underlying the impact of ethanol, focusing on neuro-astro-microglial interactions. In a relatively large collection of human iPS cells (n=36 lines) derived from subjects with extreme AUD PRSs, we hope to unravel the convergent phenotype and gene-networks that are linked to extreme high or low AUD PRSs. The results will advance our mechanistic understanding of the pathogenic role of AUD risk gene variants and the influence of polygenic risk background in a human neural system.

摘要和摘要 酒精使用障碍（AUD）是复杂的行为，伴随着大量发病，死亡率和社会 费用。遗传和环境因素都会导致AUD。尽管人类的遗传学进展 AUD，尤其是识别全基因组明显的AUD遗传风险变异的识别，AUD的神经基础 在人类中，在很大程度上是未知的。在过去的五年中，我们提供了令人信服的证据：1）人类 源自诱导多能干（IPS）细胞的神经元细胞可以用作研究的模型 神经精神疾病，包括AUD； 2）乙醇暴露会导致人类炎症体反应 神经元； 3）携带OPRM1 A118G次要基因变异的人神经元显示出对 绿酚和乙醇； 4）乙醇会导致人神经元和神经胶质细胞的基因表达变化； 5） 含小胶质细胞的3D神经元培养物可以是研究神经炎性小胶质细胞神经元的强大系统 互动。这些前提为遗传和分子的进一步机械研究奠定了基础 人类IPS细胞衍生的神经模型中AUD的基础。我们还确定了知识的关键差距 利用人类神经元作为模型系统来研究需要填充的AUD，特别是：1）乙醇如何 在人类神经膜片中影响神经胶质相互作用尚不清楚。 2）乙醇如何影响神经发生 在3D上下文中尚不清楚。此外，3）由于aud的多基因性质，单个的贡献 基因到AUD风险可能很小，表型表现受到个人通用的强烈影响 化妆品。为了解决这些悬而未决的问题，我们假设乙醇通过 影响神经相互作用，这受到单个多基因风险背景的影响。我们选择了 两个性别的36个受试者，具有极高的多基因风险评分（PR，TOP 10％ILE，n = 18 AUD）或低 PRS（底部10％Ile，n = 18，没有AUD）来检验此假设。我们将区分这些IPS细胞与这些细胞 受试者与人类星形胶质细胞共培养的2D和3D（即脑器官）神经元细胞 小胶质细胞。暴露于乙醇后，这些神经元细胞会结合形态学， 免疫细胞化学，电生理，活细胞成像和基因组分析，以揭示机制 乙醇的影响是基本的，重点是神经 - 腹部 - 神经相互作用。在一个相对较大的收藏中 来自具有极端AUD PRS的受试者的人IPS细胞（n = 36行），我们希望解开 与极高或低的AUD PRS相关的收敛表型和基因网络。结果将 促进我们对AUD风险基因变种的致病作用的机械理解和 人类神经系统中的多基因风险背景。