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.

项目摘要/摘要 通道病是由功能失调的离子通道蛋白引起的疾病或身体疾病。为了 例如，必需的机械敏感通道Piezo1和Piezo2已与多个 疾病，例如远端关节炎，脱水的遗传性气孔病和戈登综合征。那里 是在Piezo1/2中鉴定出的〜100个疾病等位基因，其中大多数引起了严重的生理 心血管，前庭，神经元和连接的组织中的疾病。尽管有电生理学 对患者细胞的研究表明，这些符号可能是由于机械转导缺陷引起的 压电疾病的基本机制或分子确定剂在很大程度上尚不清楚。在这里，我 引入一个轻松而有力的体内系统，用于对压电的功能研究；拉伸敏感和 反应性秀丽隐杆线虫生殖道。我发现功能失调的PEZO-1（唯一的直系同源 秀丽隐杆线虫中的压电会导致由于精子中的卵母细胞的碎卵母细胞而严重降低了育雏尺寸 和精子运动不良（3）。这项拟议的研究旨在发现途径和遗传的性质 使压电能够对机械刺激和坐标机械转移组织做出反应的交互器 在体内功能。此外，我将确定C中的新遗传补充剂和相关途径。 秀丽隐杆线生殖道。这项基础研究将阐明对通道病疾病的理解 由压电功能障碍和潜在的治疗药物靶靶设计引起。为了实现这些目标，我将 追求三个特定目的：第一个目的是识别秀丽隐杆线虫中PEZO-1的新遗传相互作用者。一个 遗传筛选和生化测定的组合将用于实现这一目标。我希望那个 完成提出的目标将建立秀丽隐杆线虫生殖系统作为简单而普遍的 可阐明压电生物学功能的可进行的模型，并更好地了解分子机制 PEZO-1活动。第二个目的是确定组织间信号通路是否（例如性别） 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，我将执行遗传和 生化测定以确定PEZO-1是否有助于前列腺素的合成和分泌，这 对于精子吸引力至关重要。最终目的是确定压电的目标时间和相对贡献 疾病等位基因细胞内Ca2+稳态和信号传导。为了实现这一目标，我将生成一组 组织特异性的Ca2+指标，以量化每个突变体中的钙影响。这些研究应导致 与PEZO-1相互作用的基因的全面描述以及涉及的新途径 机械转导。这项研究还将阐明遗传疾病的分子机制 由压电功能障碍引起。总体而言，这项K99/R00奖将增强我的研究技能并促进 我过渡到遗传学，发展，机械生物学和 人类稀有疾病的转化科学。