The molecular mechanism of LCD, a prevalent and conserved non-apoptotic cell death program

LCD是一种普遍且保守的非凋亡细胞死亡程序的分子机制

• 批准号: 10318927
• 负责人: Olga Yarychkivska
• 金额: $ 7.42万
• 依托单位: ROCKEFELLER UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-01-01 至 2022-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10318927
• 关键词: Adult; Apoptosis; Apoptotic; Automobile Driving; Binding; Biological Assay; Biological Models; CRISPR screen; Caenorhabditis elegans; Caspase; Cell Count; Cell Culture System; Cell Culture Techniques; Cell Death; Cells; Cessation of life; Cloaca Chamber; Computer Analysis; Cullin Proteins; Cultured Cells; Data; Development; Disease; Embryo; Event; Exhibits; Fellowship; Fibroblasts; Fingers; Genes; Genetic; Genetic Epistasis; Goals; Gonadal structure; Huntington Disease; Hybrids; Infarction; Invertebrates; Kinetics; Knowledge; Lead; Light; Lobular Neoplasia; Malignant Neoplasms; Mammalian Cell; Methods; Microscopy; Mitogen-Activated Protein Kinase Kinases; Molecular; Molecular Target; Morphology; Mus; Mutant Strains Mice; Nerve Degeneration; Organ; Pathway interactions; Patients; Peptide Hydrolases; Positioning Attribute; Proteins; Regulation; Research; Research Design; Role; Staurosporine; System; Testing; Time; Tissues; Training; Ubiquitin; Vertebrates; WNT Signaling Pathway; Yeasts; axonal degeneration; base; cell type; clinically significant; design; human disease; innovation; male; mouse model; multicatalytic endopeptidase complex; novel; novel marker; novel therapeutics; polyglutamine; polyglutamine neurodegenerative diseases; programs; protein aggregation; sperm cell; therapeutic development; transcription factor; ubiquitin-protein ligase; whole genome

Project Summary/ Abstract The long-term goal of the proposed research is to uncover molecular mechanism driving non-apoptotic cell death in vertebrate development and disease. Programed cell death functions to sculpt organs, remodel tissues, regulate cell number, and remove defective cells. While apoptosis is the most studied type of cell death, it does not account for all cellular destruction during development. Studies in C. elegans have uncovered a novel developmental cell death program, referred to as linker cell-type death (LCD), which is morphologically and molecularly distinct from apoptosis. Cell death with LCD features is commonly observed during vertebrate development and in patients and mouse models of neurodegenerative polyglutamine diseases. This proposal seeks to identify molecular events driving LCD in C. elegans, and to test their functional conservation in vertebrate settings. To rigorously investigate these mechanisms, this F32 proposal combines training in genetics, microscopy and computational analysis in vertebrate and invertebrate model systems to investigate the following specific aims: 1) identify molecular targets of the Ubiquitin Proteasome System (UPS) that precipitate cell destruction during LCD in C. elegans by performing functional studies and a yeast 2-hybrid screen; 2) uncover the molecular basis of LCD in vertebrates by developing a cell culture assay, testing roles of homologs of C. elegans LCD genes, and conducting a whole-genome CRISPR/Cas9 screen in cultured mammalian cells. The studies proposed here will lead to the development of new markers to distinguish among different types of cell death in vertebrate development and disease, while uncovering the molecular underpinnings of LCD. Because LCD is a prevalent type of cell demise, this proposal may not only shed light on basic aspects of development, neurodegeneration, and cancer, but could also eventually uncover in-roads of clinical significance.

项目总结/摘要 该研究的长期目标是揭示驱动非凋亡细胞的分子机制。 脊椎动物发育和疾病中的死亡。衰老的细胞死亡功能塑造器官，重塑 组织，调节细胞数量，并去除缺陷细胞。虽然细胞凋亡是研究最多的细胞类型， 死亡，它不能解释发育过程中所有的细胞破坏。C.优雅的有 发现了一种新的发育细胞死亡程序，称为连接细胞型死亡（LCD）， 在形态和分子上与凋亡不同。通常观察到具有LCD特征的细胞死亡 在脊椎动物发育期间以及在神经变性聚谷氨酰胺的患者和小鼠模型中 疾病该提案旨在确定C中驱动LCD的分子事件。为了测试它们的 脊椎动物环境中的功能保护。为了严格研究这些机制，F32提案 结合了遗传学，显微镜和脊椎动物和无脊椎动物模型计算分析的培训 系统，以研究以下具体目标：1）鉴定泛素蛋白酶体的分子靶标 系统（UPS），沉淀细胞破坏液晶显示器在C。通过进行功能研究， 酵母双杂交筛选; 2）通过开发细胞培养物揭示脊椎动物LCD的分子基础 分析，测试C.秀丽隐杆线虫LCD基因，并进行全基因组CRISPR/Cas9 在培养的哺乳动物细胞中筛选。本文提出的研究将导致新标记物的开发 区分脊椎动物发育和疾病中不同类型的细胞死亡， 液晶显示器的分子基础由于LCD是一种流行的电池消亡类型，因此该提议可能不会 它不仅揭示了发育、神经退行性变和癌症的基本方面，而且最终也可能 发现具有临床意义的进展