The function and regulation of the C. elegans Haspin histone kinase homolog, HASP-1

线虫 Haspin 组蛋白激酶同源物 HASP-1 的功能和调节

• 批准号: 10792737
• 负责人: David J Wynne
• 金额: $ 40.85万
• 依托单位: UNIVERSITY OF PORTLAND
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-22 至 2026-09-20
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10792737
• 关键词: Address; Animals; Binding; Biological; CRISPR/Cas technology; Caenorhabditis elegans; Cancer Model; Cancer cell line; Cell Culture Techniques; Cell Cycle; Cell division; Cells; Chromosome Segregation; Chromosomes; Clinical; Clustered Regularly Interspaced Short Palindromic Repeats; Collaborations; Complex; Conflict (Psychology); Congenital Abnormality; Defect; Down Syndrome; Drug Targeting; Ensure; Failure; Genes; Genetic; Genome; Goals; Hand; Haspin; Health; Hermaphroditism; Histone H3; Homologous Gene; Human; Infertility; Inherited; Institution; Investigation; Literature; Malignant Neoplasms; Meiosis; Mitosis; Mitotic Checkpoint; Molecular; Molecular Conformation; Monitor; Morphology; Mus; Mutation; Normal Cell; Oocytes; Organism; Pathway interactions; Pharmaceutical Preparations; Phosphorylation; Phosphotransferases; Play; Postbaccalaureate; Process; Protamine Kinase; Protein Kinase; Proteins; Recurrence; Regulation; Reporting; Research; Research Personnel; Role; Signal Transduction; Somatic Cell; Specific qualifier value; Spermatocytes; Spermatogenesis; System; Testing; Therapeutic; Time; Tissues; Toxic effect; Training; Work; Xenograft Model; anticancer activity; biomedical scientist; cancer cell; cell type; chemotherapy; chromosome missegregation; cohesion; drug development; experimental study; germline stem cells; in vivo; inhibitor; inhibitor therapy; insight; knock-down; model organism; next generation; prevent; programs; protein complex; recruit; refractory cancer; segregation; side effect; sperm cell; targeted treatment; tissue/cell culture; tool; undergraduate student

Project Summary/Abstract The process of passing on chromosomes when cells divide must be carefully monitored because errors cause birth defects and infertility and are a hallmark of cancer. Cells use a variety of mechanisms to monitor the process of chromosome segregation and these mechanisms are often the target of drugs being developed in the hopes of finding chemotherapeutics that will kill cancer cells while having low toxicity to normal cells. The Chromosomal Passenger Complex (CPC) is a protein complex that monitors chromosome segregation. To provide its essential functions, the CPC must be recruited onto chromosomes at the right place and time. The mechanisms that target the CPC to chromosomes are not fully understood, but two key pathways have been discovered that rely on the activity of the protein kinases Haspin and Bub1. While the kinase Bub1 has been well studied since its discovery as part of the mitotic checkpoint signal, Haspin is a newly discovered kinase and fundamental questions abouts its function and regulation have not yet been answered. Better understanding Haspin’s function is critical because chemotherapy drugs targeting Haspin have shown promise. Haspin has features that make its inhibitors less likely to produce side effects, and multiple inhibitors have recently shown efficacy against a variety of cancer models and low toxicity to normal cells. Investigations into Haspin’s function have so far been limited to a small number of cell types undergoing mitosis in cell culture, which offers only a partial understanding of this molecule’s function. This proposal develops a new system to study Haspin function in a variety of cell types in vivo using the model organism C. elegans, which features powerful genetic and cell biological tools. Conditional knockdown of the C. elegans Haspin protein, HASP-1, combined with specific mutations in HASP-1 and associated proteins using CRISPR gene editing, will enable us to accomplish the following Specific Aims: 1) Determine the relative contribution of the Haspin and Bub1 pathways to CPC recruitment in different cell types in C. elegans, 2) Identify the mechanism by which HASP-1 is recruited to chromosomes, and 3) Determine the mechanisms by which HASP-1 becomes activated during mitosis and meiosis. Our experimental system allows us to address these Specific Aims in multiple cell types and in meiosis, a specialized cell division where these types of mechanistic studies are typically not possible in other animal systems. The proposed work will provide new insights into the molecular mechanisms underlying the function of this important cell cycle regulator, which will positively impact human health, especially as Haspin inhibitors show up more in clinical use. Proposed experiments will be performed in close collaboration with undergraduate researchers and a post-baccalaureate researcher, enhancing the research capacity of our institution and training the next generation of biomedical scientists.

项目总结/摘要 细胞分裂时传递染色体的过程必须仔细监测， 导致出生缺陷和不孕不育，是癌症的标志。细胞使用多种机制来监控 染色体分离的过程和这些机制通常是正在开发的药物的目标 希望找到既能杀死癌细胞又对正常细胞毒性低的化疗药物。的 染色体乘客复合物（CPC）是一种蛋白质复合物，监测染色体分离。到 为了提供其基本功能，CPC必须在正确的位置和时间被招募到染色体上。的 CPC靶向染色体的机制尚未完全了解，但已经研究了两条关键途径。 发现依赖于蛋白激酶Haspin和Bub 1的活性。虽然激酶Bub 1已经被 Haspin是一种新发现的激酶，自从作为有丝分裂检查点信号的一部分被发现以来，已经得到了很好的研究 其功能和调节等基本问题尚未得到解答。更好 了解Haspin的功能是至关重要的，因为靶向Haspin的化疗药物已经显示出前景。 Haspin具有使其抑制剂不太可能产生副作用的特性，并且多种抑制剂具有 最近显示出对多种癌症模型的功效和对正常细胞的低毒性。 到目前为止，对Haspin功能的研究仅限于少数细胞类型， 细胞培养中的有丝分裂，这只提供了对这种分子功能的部分了解。这项建议 开发了一种新的系统，使用模式生物C在体内研究Haspin在多种细胞类型中的功能。 elegans，它具有强大的遗传和细胞生物学工具。有条件敲除C. elegans Haspin蛋白，HASP-1，使用CRISPR与HASP-1和相关蛋白中的特定突变组合 基因编辑，将使我们能够实现以下具体目标：1）确定 Haspin和Bub 1通路在C. elegans，2）识别 HASP-1被募集到染色体的机制，以及3）确定HASP-1被募集到染色体的机制， HASP-1在有丝分裂和减数分裂期间被激活。我们的实验系统使我们能够解决这些问题 在多种细胞类型和减数分裂中的特定目的，减数分裂是一种专门的细胞分裂，其中这些类型的机制 在其他动物系统中通常不可能进行研究。拟议的工作将提供新的见解， 这一重要的细胞周期调节因子的功能背后的分子机制，这将积极影响 人类健康，特别是Haspin抑制剂在临床应用中表现出更多。拟议的实验将是 与本科研究人员和一名学士后研究人员密切合作， 提高我们机构的研究能力，培养下一代生物医学科学家。