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.

项目摘要 PACS1综合征是影响幼儿的从头神经遗传疾病，具有独特的畸形面部特征，智力障碍和发展延迟。在全世界确定的所有情况下，个人都有相同的精确氨基酸取代，P.R203W，在高度保守的位置 PACS1蛋白。 PACS1（磷脂蛋白酸性簇排序蛋白1）首先通过与普罗蛋白转化酶弗林的相互作用和随后的研究确定已经揭示了其在秘书途径中的作用，尤其是跨性别网络。 PACS1 在所有动物中都是保守的，人类表达了额外的直系同源物pacs2。全部 PACS蛋白包含多个功能域，包括脂肪结合区域， PACS1综合征变体所属于的。目前，对Pacs1的最大理解功能来自培养细胞。必须解决的一个关键问题是R203W 改变PACS1功能并导致综合征症状。线虫C。秀丽隐杆线是用于研究人类疾病基因的可寓言模型。一个C. 秀丽隐杆线PACS-1基因编码具有所有保守域的蛋白质，雌激素和人类之间的结合区域特别保守（45％）与疾病变体位点相同/70％相似），R203在C中表示为R116。秀丽隐杆线。在这里，我们使用基因组编辑生成了CEPACS-1（R116W）模型。此外，使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醛虫的瘫痪作用具有抗性。与其神经元功能一致，我们还发现内源性PACS-1是在神经系统中表达。我们的秀丽隐杆线虫PACS-1（R116W）提供了第一个 PACS1综合征的种系表达模型。在此R03应用程序中，我们建议使用LOPAC化学库来完成化学复合屏幕，用于行为对我们的CEPACS1模型的影响。然后，我们将使用其他细胞生物学和功能测定。项目目标在可行性之内和R03申请的指南。结果将为发展提供信息 PACS1综合征治疗中的设计师制造典型，也有助于进一步 PACS1综合征的生理机制的表征。