Identifying potential therapeutics using an animal model for PACS1 syndrome

使用 PACS1 综合征动物模型确定潜在的治疗方法

基本信息

批准号：
10195626
负责人：
Dana T Byrd
金额：
$ 15.79万
依托单位：
UNIVERSITY OF CALIFORNIA, SAN DIEGO
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-05-01 至 2023-10-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10195626
关键词：
Address Affect Age Aldicarb Amino Acid Substitution Animal Behavior Animal Model Animals Appearance Behavioral Binding Biological Assay C-terminal Caenorhabditis elegans Cell Line Cell Nucleus Cellular biology Chemicals Child Choline Cholinesterase Inhibitors Cultured Cells DRPLA protein Development Developmental Delay Disorders Disease Drug usage Experimental Designs FDA approved Face Future Gene Expression Genes Genetic Goals Growth Guidelines Human Individual Intellectual functioning disability Invertebrates Language Libraries Membrane Proteins Modeling Morphology Movement Mutation Nematoda Nervous system structure Neurons Nucleotides Orthologous Gene Outcome PAC1 phosphatase Paralysed Pathologic Processes Pathway interactions Pharmaceutical Preparations Phenotype Physiological Positioning Attribute Proprotein Convertases Protein Sortings Proteins Resistance Role Site Sorting - Cell Movement Speech Study models Symptoms Synaptic Transmission Syndrome Testing Therapeutic Therapeutic Uses Variant Visualization Work clinical application de novo mutation design drug candidate esterase inhibitor experimental study gene function genome editing high-throughput drug screening human disease in vivo mutant neurogenetics protein expression protein transport screening small molecule libraries trans-Golgi Network

项目摘要

Project Summary PACS1 Syndrome is a de novo neurogenetic disorder affecting young children, characterized by distinct dysmorphic facial features, intellectual disability and developmental delays. In all cases identified throughout the world, individuals have the same exact amino acid substitution, p.R203W, in a highly conserved position of the PACS1 protein. PACS1 (Phosphofurin Acidic Cluster Sorting Protein 1) was first identified by its interaction with the proprotein convertase furin, and subsequent studies have revealed its role in secretary pathway, particularly the trans-Golgi network. PACS1 is conserved in all animals, and humans express an additional ortholog, PACS2. All PACS proteins contain multiple functional domains including the furin-binding region, in which the PACS1 syndrome variant resides. Currently, most understanding of PACS1 function is from cultured cells. A key question that must be addressed is how R203W changes PACS1 function and results in syndrome symptoms. The nematode C. elegans is a tractable model for studying human disease genes. A single C. elegans pacs-1 gene encodes a protein that shares all conserved domains, and the furin-binding region is especially well conserved between C. elegans and humans (45% identical/70% similar) with the disease variant site, R203, denoted as R116 in C. elegans. Here, we have generated a cePACS-1(R116W) model, using genome-editing. Additionally, using drugs to probe neuronal function, we found that pacs-1(R116W) animals are resistant to the paralyzing effect of the choline esterase inhibitor aldicarb. Consistent with its neuronal function, we also found that endogenous PACS-1 is expressed in the nervous system. Our C. elegans pacs-1(R116W) provides the first germline-expressed model of PACS1 syndrome. In this R03 application, we propose to complete a chemical compound screen, using the LOPAC chemical library, for behavioral effects on our cePACS1 model. We will then test top candidate hits using additional cell biology and functional assays. The project goal is within the feasibility and guideline of R03 application. The outcome will be informative for the development of designer-strategies in treatment of PACS1 syndrome, and also aid further characterization of physiological mechanisms underlying PACS1 syndrome.

项目概要 PACS1 综合征是一种影响幼儿的新发神经遗传性疾病，其特征是明显的畸形面部特征、智力障碍和发育迟缓。在世界各地发现的所有案例中，个人都有相同精确的氨基酸取代，p.R203W，在高度保守的位置 PACS1 蛋白。 PACS1（磷酸呋喃酸性簇分选蛋白1）是第一个通过其与前蛋白转化酶弗林蛋白酶的相互作用以及随后的研究来鉴定揭示了其在秘书通路，特别是跨高尔基体网络中的作用。 PACS1 在所有动物中都是保守的，人类表达一个额外的直系同源物，PACS2。全部 PACS 蛋白含有多个功能域，包括弗林蛋白酶结合区， PACS1 综合征变异所在。目前最了解的PACS1 功能来自培养细胞。必须解决的一个关键问题是R203W如何改变 PACS1 功能并导致综合征症状。线虫C. 线虫是研究人类疾病基因的易于处理的模型。单个C。线虫 pacs-1 基因编码一种共享所有保守结构域的蛋白质，并且弗林蛋白酶结合区在秀丽隐杆线虫和人类之间尤其保守（45% 相同/70%相似）与疾病变异位点 R203，在 C. 中表示为 R116。线虫。在这里，我们使用基因组编辑生成了 cePACS-1(R116W) 模型。此外，利用药物探测神经元功能，我们发现pacs-1(R116W) 动物对胆碱酯酶抑制剂涕灭威的麻痹作用有抵抗力。与其神经元功能一致，我们还发现内源性 PACS-1 是表达于神经系统。我们的线虫 pacs-1(R116W) 提供了第一个 PACS1 综合征种系表达模型。在此 R03 应用中，我们建议使用 LOPAC 化学库完成化合物筛选，用于行为对我们的 cePACS1 模型的影响。然后，我们将使用以下方法测试最佳候选命中额外的细胞生物学和功能测定。项目目标在可行性范围内 R03应用指南。结果将为发展提供信息治疗 PACS1 综合征的设计策略，也有助于进一步 PACS1 综合征生理机制的表征。