Identifying pathogenic mechanisms underlying PACS1 Syndrome: implications for neural development

识别 PACS1 综合征的致病机制：对神经发育的影响

基本信息

批准号：
10676187
负责人：
ALICIA DIONE GUEMEZ GAMBOA
金额：
$ 53.13万
依托单位：
NORTHWESTERN UNIVERSITY AT CHICAGO
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-08-05 至 2027-07-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10676187
关键词：
3-Dimensional Address Affect Arginine Binding Bipolar Disorder Brain Cell Death Cell Line Cell Nucleus Cell Proliferation Cell physiology Cells Cerebral cortex Cerebrum Chromatin Complex Cytoplasm Data Data Set Defect Development Developmental Delay Disorders Disease Dorsal Endosomes Equilibrium Etiology FOXG1B gene Family Foundations Funding Gene Expression Generations Genes Genetic Genetic Transcription Glutamates Goals Histone Deacetylase Human Impaired cognition Individual Knowledge Label Lentivirus Location Mediating Modernization Molecular Molecular Biology Nervous System Neurodevelopmental Disorder Neurologic Neuronal Differentiation Neurons Neurophysiology - biologic function Neurosciences Nuclear Nuclear Localization Signal Organoids PACS1 Syndrome Pathogenicity Pathway interactions Patients Pattern Phenotype Process Proliferating Prosencephalon Proteins Recurrence Research Role S phase SWI1 Sorting Specific qualifier value Stains Structure Susceptibility Gene Techniques Testing Tissue-Specific Gene Expression Tryptophan Variant Work cancer cell cell type design differential expression effective therapy experimental study genome wide association study induced pluripotent stem cell myocyte-specific enhancer-binding-factor 2C nerve stem cell nervous system development neural neurodevelopment neurogenesis neuromechanism programs protein transport severe early onset obesity single-cell RNA sequencing stem cells synaptogenesis targeted treatment tool trans-Golgi Network transcriptome

项目摘要

Project Summary Neurodevelopmental disorders (NDDs) are heterogenous and usually present with complex etiology. Individuals with these conditions present with cognitive impairment accompanied by lifelong deficits; yet remarkably little is known about their neurological basis. Patients and their families are in desperate need for disease-modifying therapies. However, to develop effective treatments, it is imperative to understand the dysregulation of molecular and cellular processes leading to these conditions. PACS1 Syndrome is a NDD caused by a single R203W substitution in the Phosphofurin Acidic Cluster Sorting 1 (PACS1) protein in over 200 patients. Moreover, genome-wide association studies (GWAS) identified the human PACS1 locus as a susceptibility gene in severe early-onset obesity, developmental delay, and bipolar disorder, suggesting a broader role in brain development. PACS1 encodes a multifunctional protein which canonical cytosolic function in the secretory pathway is to direct its cargo to the trans-Golgi Network.However, PACS1 also contains a nuclear localization signal (NLS), shuttles to the nucleus during S phase, and, at least in highly proliferative cancer cells, regulates chromatin stability through interaction with HDAC proteins. Despite previous studies have revealed critical PACS1 functions, the role of PACS1 within the nucleus as well as the effect of the PACS1 R203W variant in the nervous system yet to be determined. Thus, we have generated patient induced pluripotent stem cells (iPSCs)-derived cerebral organoids to investigate the transcriptome of a developing patterned, three-dimensional neural structure with the patient genetic background. By performing single cell RNA Sequencing in organoids during early development, we generated a preliminary dataset that strongly indicate that regulates a gene expression program important for Glutamatergic/GABAergic fate specification. Altogether, evidence regarding nuclear localization of PACS1, in addition to our preliminary data, suggests that PACS1 has an unexplored nuclear function, possibly by regulating gene expression in neural progenitors. Therefore, we hypothesize that PACS1 regulates Glutamatergic/GABAergic balance through a non-canonical nuclear function that decreases as neurogenesis proceeds. Thus, PACS1 deficits result in a shift towards GABAergic fate, generating ectopic GABAergic neurons, subsequent Glutamatergic/GABAergic imbalance, and finally NDD phenotypes. We will test this hypothesis by addressing whether PACS1 possesses context specific functions that differ across neural differentiation (aim1), and deficits in PACS1 nuclear function alter Glutamatergic/GABAergic specification balance (aim2). Results from this proposal will greatly enhance our understanding of how the PACS1 deficits affect the developing nervous system, by uncovering a currently disregarded nuclear function. Moreover, our work will expand our knowledge of the molecular underpinnings and consequences of the Glutamatergic/GABAergic imbalance, that has repeatedly been described as a precursor of many NDDs, pointing towards a possible common mechanism from genetically heterogeneous origins and may ultimately reveal convergent therapy targets.

项目摘要神经发育障碍（NDDS）是异质的，通常具有复杂的病因。个人在这些情况下，认知障碍伴随着终生缺陷；但是，很少有知道他们的神经依据。患者及其家人迫切需要疾病修改疗法。但是，要开发有效的治疗方法，必须了解分子的失调以及导致这些条件的细胞过程。 PACS1综合征是由单个R203W引起的NDD 在200多名患者中取代磷脂蛋白酸性分类1（PACS1）蛋白。而且，全基因组关联研究（GWAS）将人PACS1基因座确定为严重的敏感基因早期发作的肥胖，发育延迟和躁郁症，表明在大脑发育中起着更广泛的作用。 PACS1编码一种多功能蛋白，该蛋白质在分泌途径中的规范胞质功能是指导它的货物到跨高尔基网络。在S期到核，至少在高度增殖的癌细胞中调节染色质稳定性通过与HDAC蛋白的相互作用。尽管以前的研究揭示了关键的PACS1功能，但 PACS1在细胞核中的作用以及PACS1 R203W变体在神经系统中的作用要确定。因此，我们已经产生了患者引起的多能干细胞（IPSC）衍生的脑器官研究正在发展的图案化的三维神经结构的转录组患者遗传背景。通过在早期发育过程中在器官中进行单细胞RNA测序，我们生成了一个初步数据集，该数据集强烈表明调节基因表达程序很重要用于谷氨酸/GABA能命运规格。总共，关于PACS1的核定位的证据，除我们的初步数据外，还表明PACS1具有未开发的核功能，可能是调节神经祖细胞中的基因表达。因此，我们假设PACS1调节谷氨酸能/GABA能通过非经典核功能降低为神经发生收益。因此，PACS1缺陷导致向GABA能命运转移，产生异位GABA能神经元，随后的谷氨酸能/GABA能失衡，最后是NDD表型。我们将通过解决PACS1是否具有在神经分化中不同的上下文特定功能（AIM1）， PACS1核功能的缺陷改变了谷氨酸能/GABA能规格平衡（AIM2）。结果该建议将大大增强我们对PACS1缺陷如何影响发展紧张的理解系统，通过发现当前无视的核功能。此外，我们的工作将扩大我们的知识谷氨酸能/GABA能失衡的分子基础和后果反复被描述为许多NDD的前体，指出了一种可能的常见机制遗传上异质的起源，最终可能揭示了收敛疗法靶标。