Circadian Rhythms in Neuronal Models of Bipolar Disorder

双相情感障碍神经元模型中的昼夜节律

• 批准号: 10398884
• 负责人: Michael Joseph McCarthy
• 金额: --
• 依托单位: VA SAN DIEGO HEALTHCARE SYSTEM
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-04-01 至 2025-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10398884
• 关键词: ANK3 gene; Activity Cycles; Affect; Aftercare; Animal Model; Animals; Antipsychotic Agents; Biological Assay; Biological Markers; Bipolar Disorder; Bipolar Neuron; Candidate Disease Gene; Cell model; Cells; Cellular biology; Circadian Dysregulation; Circadian Rhythms; Clinical; Corpus striatum structure; Data; Diagnosis; Diagnostic; Disease; Dopamine; Dopamine D2 Receptor; Environmental Risk Factor; Extended Family; Fibroblasts; Foundations; Future; Genes; Genetic; Genetic Risk; Genetic study; Glutamates; Heritability; Human; In Vitro; Individual; Light; Link; Lithium; Maintenance Therapy; Measures; Mental disorders; Methods; Modeling; Molecular Biology; Mood Disorders; Moods; Neurons; Occupational; Ontology; Pathway Analysis; Patients; Periodicity; Pharmaceutical Preparations; Phase; Phenotype; Population; Prediction of Response to Therapy; Process; Protocols documentation; Reporter; Research; Risk; Risk Factors; Role; Sampling; Signal Transduction; Skin; Sleep; Small Interfering RNA; System; TCF7L2 gene; Temperature; Time; Veterans; Work; base; cell type; circadian; circadian pacemaker; clinical predictors; design; disability; disorder control; effective therapy; excitatory neuron; experience; genome wide association study; induced pluripotent stem cell; insight; knock-down; loss of function; member; mesolimbic system; molecular clock; mood symptom; nerve stem cell; novel; polygenic risk score; preference; response; risk variant; severe mental illness; social; stem cell model; suicide rate; transcriptome; transcriptome sequencing

Bipolar disorder (BD) is a psychiatric disorder associated with heritable polygenic risk factors. While clinically heterogeneous, key clinical features of BD include disruptions in daily sleep and activity cycles indicating that circadian rhythm disruption is an important part of the disorder in many patients. Circadian rhythms are determined by molecular clocks comprised of “clock genes” whose expression is regulated by environmental factors such as light and temperature. In recent years, our group and others have made progress in developing cellular models of BD using induced pluripotent stem-cell (iPSC) derived neuronal progenitor cells (NPCs) and neurons. We have determined that chronotype (time of day preference, “morningness”) in BD patients and circadian rhythms in cells both predict the clinical response to lithium maintenance therapy. Using iPSC-based methods to grow human NPCs and neurons, we found that neurons from BD patients have a hyperexcitable phenotype that can be reversed by lithium selectively in neurons from lithium-responsive (Li-R) BD donors, but not in cells from lithium non-responders (Li-NR). Presently, we propose to build upon this work and further develop the iPSC-neuron model to further investigate circadian disruption in BD. In Aim 1, we will estimate the contributions of polygenic risk factors to circadian rhythm phenotypes in BD in excitatory neurons. We will study rhythms in iPSC-derived NPC and glutamatergic neurons grown from a well-characterized, extended family who share high genetic risk for BD, but are discordant for the BD diagnosis. Presumably, cells from these genetically similar individuals will contain BD risk genes across a gradient that can be quantified using polygenic risk scores (PRS). By evaluating the relationship between PRS and rhythm parameters, we will estimate the aggregate contributions of genetic risk for BD to the expression of circadian rhythm abnormalities. In Aim 2 we propose to assess the contribution of BD-associated gene sets to circadian rhythms in neurons. Genome-wide association studies (GWAS) of sleep and circadian phenotypes suggest the existence of a shared genetic basis for mood disorders and chronotype. In this aim, we will study synchronized cells over a 24 h period to identify rhythmically expressed genes in neural progenitor cells (NPCs) from BD patients and controls and describe rhythm differences between diagnostic groups. We expect to find that many genes change their rhythm or lose rhythms altogether in BD neurons. We will then use a molecular biology method called siRNA knockdown to reduce the expression of candidate genes linked to BD and the circadian clock, and assess their loss of function in circadian rhythm assays, under constant or temperature-entrained conditions. In Aim 3 we propose to measure circadian rhythms in iPSC-derived GABAergic medium-spiny neuron-like cells. Our previous cell-based studies of BD employed, excitatory glutamatergic neurons. Animal models of BD show important contributions of the clock genes on dopamine projections to inhibitory, GABAergic medium spiny neurons (MSN) in the striatum. In this aim, we will differentiate iPSCs from BD patients (Li-R and Li-NR) into inhibitory, GABAergic MSN-like neurons using an established, high efficiency protocol. We will use the Per2-luc circadian rhythm reporter to measure rhythms in live human GABAergic neurons for the first time and assess the effects of mood stabilizing drugs such as lithium and antipsychotic drugs upon them. At the conclusion of the study, we will have a more advanced understanding of which neurons lose rhythms in BD, which BD-risk genes are involved in the circadian disruption, and how circadian disruption affects the function of neurons. This information may help to better identify lithium- responsive BD patients and link them efficiently to effective treatments.

躁郁症（BD）是一种与可遗传的多基因危险因素相关的精神病。而诊所 BD的异构，主要的临床特征包括日常睡眠和活动周期的中断，表明 在许多患者中，昼夜节律破坏是该疾病的重要组成部分。昼夜节律是 由“时钟基因”的分子钟确定，其表达受环境调节 光和温度等因素。近年来，我们的小组和其他人在发展方面取得了进步 使用诱导多能干细胞（IPSC）衍生的神经元祖细胞（NPC）和 神经元。我们已经确定BD患者的计时型（日期偏好，“早晨”）和 细胞中的昼夜节律都预测了对锂维持疗法的临床反应。使用基于IPSC的 种植人NPC和神经元的方法，我们发现来自BD患者的神经元可过度 锂可以从锂反应（LI-R）BD供体中选择性地反转锂的表型，但是 不在锂非反应器（LI-NR）中的细胞中。目前，我们建议以这项工作为基础 开发IPSC-Neuron模型，以进一步研究BD中的昼夜节律破坏。在AIM 1中，我们将估计 多基因风险因素对兴奋性神经元中BD中昼夜节律表型的贡献。我们将学习 IPSC衍生的NPC和谷氨酸能神经元的节奏是从一个特征良好的大家庭中种植的 具有BD的高遗传风险，但对于BD诊断是不一致的。据推测，这些细胞从遗传上 相似的个体将在梯度上包含BD风险基因，该梯度可以使用多基因风险评分进行量化 （PR）。通过评估PRS和节奏参数之间的关系，我们将估算骨料 BD遗传风险对昼夜节律异常表达的贡献。在目标2中，我们建议 评估BD相关基因集对神经元中昼夜节律的贡献。全基因组关联 对睡眠和昼夜节律表型的研究（GWAS）表明存在共同的情绪遗传基础 疾病和表型。在此目标中，我们将在24小时内研究同步细胞，以节奏地识别 来自BD患者和对照的神经祖细胞（NPC）中表达的基因并描述节奏 诊断组之间的差异。我们希望发现许多基因改变了节奏或失去节奏 在BD神经元中总共。然后，我们将使用称为siRNA敲低的分子生物学方法来减少 与BD和昼夜节律相关的候选基因的表达，并评估它们在昼夜节律中的功能丧失 节奏测定，在恒定或温度进入条件下。在AIM 3中，我们建议测量昼夜节律 IPSC衍生的GABA能中螺旋神经元样细胞中的节奏。我们以前基于细胞的BD研究 使用的兴奋性谷氨酸能神经元。 BD的动物模型显示了时钟的重要贡献 多巴胺上的基因针对纹状体中的抑制性，GABA能培养基神经元（MSN）。在这个 目的，我们将IPSC与BD患者（LI-R和LI-NR）区分为抑制性GABA能MSN样神经元 使用建立的高效率协议。我们将使用PER2-LUC昼夜节律记者进行测量 首次活着的人类GABA能神经元中的节奏和评估情绪稳定药物的影响 例如锂和抗精神病药。在研究结束时，我们将有一个更高级的 了解哪些神经元在BD中失去了节奏，BD风险基因参与昼夜节律的破坏， 以及昼夜节律的破坏如何影响神经元的功能。这些信息可能有助于更好地识别锂 反应性的BD患者并将其有效地与有效治疗联系起来。