Modeling PIEZO associated diseases in Caenorhabditis elegans: from genetics to mechanism

秀丽隐杆线虫 PIEZO 相关疾病建模：从遗传学到机制

基本信息

批准号：
10866791
负责人：
Xiaofei Bai
金额：
$ 24.61万
依托单位：
UNIVERSITY OF FLORIDA
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：
2023-08-01 至 2026-07-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10866791
关键词：
Address Affect Alleles Animal Model Arthrogryposis Award Basic Science Biochemical Biological Assay Biological Process Caenorhabditis elegans Calcium Calcium Signaling Cardiovascular system Cells Connective Tissue Data Defect Dehydration Development Disease Distal Drosophila genus Drug Targeting Dysplasia Electrophysiology (science)Exocytosis Functional disorder Genes Genetic Genetic Predisposition to Disease Genetic Screening Goals Gonadal Steroid Hormones Gonadal structure Gordon syndrome Hermaphroditism Homeostasis Homologous Gene Human Image Inherited Ion Channel Protein Knowledge Link Mass Spectrum Analysis Membrane Missense Mutation Modeling Molecular Mus Mutation Nature Nematoda Neurons Oocytes Orthologous Gene Pathway interactions Patients Pharyngeal structure Phenotype Physiological Physiological Processes Piezo 1 ion channel Piezo 2 ion channel Prostaglandins Pump Rare Diseases Regulation Reproduction Reproductive Process Reproductive system Research Research Personnel Role Series Signal Pathway Signal Transduction Sperm Motility Stretching Suppressor Mutations Symptoms System Testing Therapeutic Tissues Translational Research Variant Work Zebrafish career design egg gene interaction genetic analysis genetic approach genome sequencing in vivo mechanical stimulus mechanotransduction mutant new therapeutic target novel novel therapeutics reproductive tract sensor skills sperm cell stomatocytic anemia trafficking whole genome

项目摘要

Project Summary/ Abstract Channelopathies are diseases or physiological disorders caused by the dysfunctional ion channel proteins. For example, the essential mechanosensitive channels PIEZO1 and PIEZO2 have been tightly linked to multiple diseases, such as distal arthrogryposis, dehydrated hereditary stomatocytosis, and Gordon Syndrome. There are ~100 disease alleles that have been identified in PIEZO1/2, most of which caused severe physiological disorders in cardiovascular, vestibular, neuronal, and connective tissues. Despite the electrophysiological studies in the patients’ cells indicated that these symptoms are likely due to a mechanotransduction defect, the underlying mechanisms or molecular determinants of PIEZO diseases remain largely unknown. Here, I introduce a facile and powerful in vivo system for the functional study of PIEZO; the stretch sensitive and responsive C. elegans reproductive tract. I have discovered that the dysfunctional PEZO-1 (the sole ortholog of PIEZO in C. elegans) causes severely reduced brood sizes due to the crushing oocytes in the spermatheca and poor sperm motility (3). This proposed study aims to discover the nature of the pathways and genetic interactors that enable PIEZO to respond to mechanical stimuli and coordinate mechanotransductive tissue function in vivo. Furthermore, I will identify new genetic suppressors and associated pathways in the C. elegans reproductive tract. This basic research will shed light on the understanding of channelopathy diseases caused by PIEZO dysfunction and the potential therapeutical drug target design. To achieve these goals, I will pursue three specific aims: The first aim is to identify novel genetic interactors of PEZO-1 in C. elegans. A combination of genetic screens and biochemical assays will be used to achieve this aim. I expect that completing the proposed aims will establish the C. elegans reproductive system as a simple and genetically tractable model to elucidate PIEZO biological functions and to better understand the molecular mechanisms of PEZO-1 activity. The second aim is to determine whether inter-tissue signaling pathways (such as the sex hormone prostaglandin) is affected in pezo-1 mutants. To achieve this aim, I will perform genetic and biochemical assays to determine whether PEZO-1 contributes to prostaglandin synthesis and secretion, which are essential for sperm attraction. The final aim is to identify target tissues and relative contribution of PIEZO disease alleles to intracellular Ca2+ homeostasis and signaling. To achieve this aim, I will generate a set of the tissue-specific Ca2+ indicators to quantify the calcium influx in each mutant. These studies should lead to a comprehensive delineation of genes that interact with pezo-1, and new pathways that involve mechanotransduction. This research will also shed light on the molecular mechanisms of the genetic diseases caused by PIEZO dysfunction. Overall, this K99/R00 award will strengthen my research skillset and facilitate my transition into an independent researcher in the field of genetics, development, mechanobiology, and translational science of human rare diseases.

项目概要/摘要离子通道病是由功能失调的离子通道蛋白引起的疾病或生理障碍。例如，重要的机械敏感通道 PIEZO1 和 PIEZO2 已与多个紧密相连疾病，例如远端关节弯曲、脱水遗传性口细胞增多症和戈登综合征。 PIEZO1/2 中已鉴定出约 100 个疾病等位基因，其中大多数会导致严重的生理症状尽管存在电生理学方面的疾病，但心血管、前庭、神经元和结缔组织的疾病。对患者细胞的研究表明，这些症状可能是由于机械传导缺陷造成的，即 PIEZO 疾病的潜在机制或分子决定因素仍然很大程度上未知。引入一个简单而强大的体内系统，用于 PIEZO 的功能研究；我发现了功能性的 PEZO-1（唯一的直系同源物）。线虫中的压电（PIEZO）由于受精囊中的卵母细胞被压碎而导致幼崽尺寸严重减小和精子活力差 (3)。这项拟议的研究旨在发现途径和遗传的本质。使 PIEZO 能够响应机械刺激并协调机械传导组织的交互器此外，我将在 C 中鉴定新的基因抑制因子和相关途径。这项基础研究将有助于了解线虫生殖道疾病。由PIEZO引起的功能障碍和潜在的治疗药物靶点设计，我将实现这些目标。追求三个具体目标：第一个目标是鉴定秀丽隐杆线虫中 PEZO-1 的新遗传相互作用因子。我预计将结合遗传筛选和生化检测来实现这一目标。完成拟议的目标将使线虫生殖系统成为一个简单的遗传系统易于处理的模型来阐明 PIEZO 的生物学功能并更好地理解其分子机制 PEZO-1 活性的第二个目的是确定组织间信号通路（例如性别）是否存在。激素前列腺素）在 pezo-1 突变体中受到影响为了实现这一目标，我将进行遗传和基因检测。生化测定以确定 PEZO-1 是否有助于前列腺素合成和分泌，这对于精子吸引至关重要，最终目的是确定目标组织和 PIEZO 的相对贡献。细胞内 Ca2+ 稳态和信号传导的疾病等位基因为了实现这一目标，我将生成一组组织特异性 Ca2+ 指标来量化每个突变体中的钙流入。这些研究应该得出一个结果。全面描述与 pezo-1 相互作用的基因，以及涉及的新途径这项研究还将揭示遗传疾病的分子机制。总的来说，这个 K99/R00 奖项将增强我的研究技能并促进我的研究技能。我转变为遗传学、发育学、机械生物学等领域的独立研究员人类罕见疾病的转化科学。