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

• 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
• 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

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.

项目总结/摘要 离子病是由功能失调的离子通道蛋白引起的疾病或生理障碍。为 例如，基本的机械敏感性通道PIEZO 1和PIEZO 2已经与多个 疾病，如远端关节弯曲、脱水遗传性口细胞增多症和戈登综合征。那里 在PIEZO 1/2中已经鉴定了约100种疾病等位基因，其中大多数引起严重的生理性疾病。 心血管、前庭、神经元和结缔组织疾病。尽管电生理 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， PIEZO疾病的潜在机制或分子决定因素在很大程度上仍然未知。这里我 介绍了一个简单而强大的体内系统，用于PIEZO的功能研究; 响应C.线虫生殖道我发现功能失调的PEZO-1（PEZO-1的唯一直系同源物） PIEZO in C.由于受精囊中的卵母细胞被压碎， 精子活力差（3）。这项拟议的研究旨在发现途径的性质和遗传 使PIEZO能够响应机械刺激并协调机械传导组织的相互作用物 在体内发挥作用。此外，我将确定新的遗传抑制因子和相关的途径，在C。 线虫生殖道这一基础研究将有助于了解通道病疾病 由PIEZO功能障碍引起的潜在治疗药物靶点设计。为了实现这些目标，我将 本研究有三个具体目标：第一个目标是鉴定PEZO-1在C.优雅的。一 将采用遗传筛选和生化分析相结合的方法来实现这一目标。我预计 完成拟议的目标将建立C。线虫的生殖系统是一个简单的， 易于处理的模型，以阐明PIEZO的生物学功能，并更好地了解分子机制， PEZO-1活性。第二个目的是确定组织间的信号通路（如性别）是否 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 生物化学测定以确定PEZO-1是否有助于前列腺素的合成和分泌， 是吸引精子的关键最终目的是确定靶组织和PIEZO的相对贡献 疾病等位基因对细胞内Ca 2+稳态和信号传导的影响。为了实现这个目标，我将生成一组 组织特异性Ca 2+指标，以量化每个突变体中的钙内流。这些研究将导致 全面描绘与pezo-1相互作用的基因，以及涉及 机械传导这项研究还将揭示遗传性疾病的分子机制 是由PIEZO功能障碍引起的总的来说，这个K99/R 00奖将加强我的研究技能，并促进 我转变为一个独立的研究人员在遗传学，发展，机械生物学领域， 人类罕见疾病的转化科学。