Common schizophrenia variants functioning in developmental human cortical interneurons

在发育中的人类皮质中间神经元中发挥作用的常见精神分裂症变异

基本信息

批准号：
10735990
负责人：
SANGMI CHUNG
金额：
$ 81.27万
依托单位：
NEW YORK MEDICAL COLLEGE
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-07-01 至 2028-04-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10735990
关键词：
3-Dimensional ATAC-seq Address Adult Affect Autopsy Biological Brain CRISPR interference CRISPR-mediated transcriptional activation Cells ChIP-seq Chromatin Chromosomes Code Complement Complex Data Data Set Development Developmental Process Disease Distal Early identification Embryo Embryonic Development Fetal Development Gene Expression Gene Expression Profile Generations Genes Genetic Genetic Risk Genetic study Genome Genomics Genotype Glean Goals Heritability Human In Vitro Interneurons Libraries Maps Medial Methods Mus Neurodevelopmental Disorder Neurons Parvalbumins Pathologic Pathway interactions Patients Pluripotent Stem Cells Population Procedures Qualifying Quantitative Trait Loci Regulation Regulator Genes Resolution Role Schizophrenia Somatostatin Synapses Testing Time Tissues Training Transplantation Untranslated RNA Validation Variant Weight Work brain cell brain tissue cell type cohort fetal gene regulatory network genetic makeup genome wide association study genome-wide human fetal brain in vivo induced pluripotent stem cell insight migration new therapeutic target risk variant schizophrenia risk stem cell differentiation transcription factor transcriptome transcriptome sequencing transcriptomics

项目摘要

Abstract Schizophrenia (SCZ) is a highly heritable and complex neurodevelopmental disorder. Remarkable advances have been made recently in SCZ genetic studies with an increasing number of risk loci reaching genome-wide significance; however, gleaning biological insight from these loci has been challenging. The majority of SCZ risk loci are located in non-coding regions. As such, it is hypothesized that they function by regulating distal gene expression via 3D chromatin interactions. However, it has yet to be determined which loci are operational in which cells, at what time points, and with what impact. Recent genomic analyses showed enriched SCZ heritability in human fetal brains rather than adult brains, suggesting the role of SCZ risk loci in modulating fetal development for increased SCZ risks. Thus, unraveling SCZ risk loci function during development will be critical for understanding genetic influences on SCZ risks. Genetic influences on gene expression (e.g. expression quantitative trait loci (eQTLs)) are cell-type-specific, and sometimes confer opposing effects depending on the cell type, underscoring the importance of cell-type-specific studies using homogeneous cell populations for a clear mechanistic understanding. Parvalbumin (PV)- or somatostatin (SST)-expressing medial ganglionic eminence (MGE)-derived cortical interneurons (cINs) are consistently affected in SCZ brains. More importantly, SCZ heritability is shown to be enriched in MGE cells in human fetal brains, necessitating the study of these cells to understand the mechanisms of SCZ risk loci. Although there are no postmortem fetal SCZ tissues for mechanistic study, in vitro differentiation of iPSC—which well recapitulates early embryonic development— provides developmental SCZ brain cells with the same genetic makeup as patient brains. We established methods for the efficient generation of homogeneous populations of MGE-derived cINs from healthy control (HC) and SCZ iPSCs. We also extensively validated their functionality and authenticity both in vitro and in vivo, including robust migration and synaptic integration into host brains that results in efficient inhibitory regulation of host circuitry in transplanted mice. Using an unprecedentedly large number of iPSCs to provide homogeneous populations of HC vs SCZ fetal cINs for mechanistic studies, we will address our hypothesis that SCZ risk loci active in developmental MGE-type cINs regulate distal gene expression via 3D chromatin interactions. Employing transcriptome analysis, PrediXcan analysis, and Micro-C analysis, we will map SCZ risk loci with unknown functions to the risk genes they regulate in these vulnerable cell populations during development. Developmental cIN-specific genetic influences on gene expression, identified based on multiple lines of corroborating evidence, will be functionally validated using CRISPRi/CRISPRa approaches. This unbiased genome-wide comprehensive data set from developmental MGE-type cINs with functional validation will provide a road map for unravelling the genetic basis of developmental SCZ risks and help us identify mechanism-based novel therapeutic targets.

抽象的精神分裂症（SCZ）是一种高度遗传且复杂的神经发育障碍。最近在 SCZ 遗传研究中进行了越来越多的风险基因座达到全基因组重要性；然而，从这些位点收集生物学见解一直具有挑战性。位点区域位于非编码区，因此，它们通过调节远端基因发挥作用。通过 3D 染色质相互作用表达然而，尚未确定哪些基因座在其中起作用。最近的基因组分析显示了哪些细胞、在什么时间点以及有什么影响。人类胎儿大脑而不是成人大脑的遗传力，表明 SCZ 风险位点在调节胎儿大脑中的作用因此，在开发过程中解开 SCZ 风险位点的功能至关重要。用于了解遗传对 SCZ 风险的影响遗传对基因表达的影响（例如数量性状位点 (eQTL) 是细胞类型特异性的，有时会根据细胞类型产生相反的效果细胞类型，强调使用同质细胞群进行细胞类型特异性研究的重要性清楚地了解表达小白蛋白（PV）或生长抑素（SST）的内侧神经节。更重要的是，SCZ 大脑中的高位 (MGE) 衍生的皮质中间神经元 (cIN) 始终受到影响。 SCZ 遗传力在人类胎儿大脑的 MGE 细胞中富集，因此有必要对这些进行研究尽管没有死后胎儿 SCZ 组织，但我们仍无法利用细胞来了解 SCZ 风险位点的机制。机制研究，iPSC 的体外分化——很好地概括了早期胚胎发育—— 提供与患者大脑具有相同基因组成的发育性 SCZ 脑细胞。从健康对照中有效产生同质 MGE 衍生 cIN 群体的方法 (HC) 和 SCZ iPSC 还在体外和体内广泛验证了它们的功能和真实性，包括强大的迁移和突触整合到宿主大脑中，从而产生有效的抑制调节使用前所未有的大量 iPSC 来提供同质的移植小鼠的宿主电路。 HC 与 SCZ 胎儿 cIN 群体的机制研究，我们将提出我们的假设，即 SCZ 风险位点活跃于发育中的 MGE 型 cIN 通过 3D 染色质相互作用调节远端基因表达。利用转录组分析、PrediXcan 分析和 Micro-C 分析，我们将绘制 SCZ 风险位点它们在发育过程中在这些脆弱细胞群中调节的风险基因的功能未知。基于多个品系鉴定的发育 cIN 特异性遗传对基因表达的影响确凿的证据将使用 CRISPRi/CRISPRa 方法进行功能验证。来自发育中的 MGE 型 cIN 的全基因组综合数据集以及功能验证将提供阐明发育性 SCZ 风险的遗传基础并帮助我们识别基于机制的路线图新的治疗靶点。