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

基本信息

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
项目类别：
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

项目摘要

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.

项目摘要/摘要当必须仔细监测细胞分裂时，传递染色体的过程是因为错误导致先天缺陷和不育，是癌症的标志。细胞使用多种机制来监测染色体分离和这些机制的过程通常是正在开发的药物的目标希望找到化学治疗剂，可以杀死癌细胞，而对正常细胞的毒性低。染色体乘客复合物（CPC）是一种监视染色体分离的蛋白质复合物。到提供其基本功能，必须将CPC招募到正确的位置和时间的染色体上。这将CPC靶向染色体的机制尚不完全了解，但是已经有两个关键途径发现依赖于蛋白激酶haspin和bub1的活性。激酶Bub1是自从发现作为有丝分裂检查点信号的一部分以来，Haspin是一种新发现的激酶尚未回答有关其功能和法规的基本问题。更好的了解Haspin的功能至关重要，因为靶向Haspin的化学疗法药物已经表现出了希望。 Haspin具有使其抑制剂产生副作用的可能性较小的功能，并且多种抑制剂具有最近显示出针对多种癌症模型的效率和对正常细胞的低毒性。到目前为止，对Haspin功能的调查仅限于正在进行的少数细胞类型细胞培养中的有丝分裂仅提供对该分子功能的部分理解。这个建议使用模型有机体C开发了一个新的系统，用于研究体内各种细胞类型的HASPIN功能。秀丽隐杆线，具有强大的遗传和细胞生物学工具。秀丽隐杆线虫的有条件击倒 HASPIN蛋白HASP-1与HASP-1和相关蛋白的特异性突变结合使用CRISPR 基因编辑，将使我们能够完成以下特定目标：1）确定秀丽隐杆线虫中不同细胞类型中CPC募集的Haspin和Bub1途径，2）确定将HASP-1募集到染色体的机制，以及3）确定的机制 HASP-1在有丝分裂和减数分裂过程中被激活。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是专门的细胞分裂在其他动物系统中，通常无法进行研究。拟议的工作将为您提供新的见解该重要细胞周期调节剂功能的基础的分子机制，这将积极影响人类健康，尤其是Haspin抑制剂在临床使用方面的表现更多。建议的实验将是与本科研究人员和学士后的研究人员密切合作，增强我们机构的研究能力，并培训下一代生物医学科学家。