Microtubule dynamics and error correction

微管动力学和误差校正

基本信息

批准号：
10640162
负责人：
Linda Wordeman
金额：
$ 37.93万
依托单位：
UNIVERSITY OF WASHINGTON
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-06-07 至 2026-02-28
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10640162
关键词：
ATP phosphohydrolase Antineoplastic Agents Biological Assay Brain CRISPR/Cas technology Cell Culture Techniques Cell Line Cell Survival Cell division Cells Centrosome Chromosomal Instability Chromosome Segregation Chromosomes Classification Clustered Regularly Interspaced Short Palindromic Repeats Congenital Abnormality Congresses Coupled Development Embryo Embryonic Development Engineering Enhancers Enzymatic Biochemistry Fluorescence Fluorescence Microscopy Functional disorder Future Generations Genome Geometry Goals Human Image Imaging Device Impairment Investigation Kinesin Kinetochores Length MXD1 gene Malignant Neoplasms Microscopy Microtubules Mitosis Mitotic Motor Pathway interactions Pharmaceutical Preparations Pharmacology Phosphotransferases Positioning Attribute Predisposition Proteasome Inhibitor Proteins Resistance Role Sirolimus Testing Therapeutic Transgenic Organisms Tubulin Vertebrates Zebrafish cell fixing cohesion embryo cell hatching knowledge base mutant nerve stem cell neurodevelopment new therapeutic target overexpression premature preservation prevent public health relevance single molecule small molecule stem cell division stem cells targeted agent therapeutic target tumorigenic

项目摘要

Modified Project Summary/Abstract Section The broad goal of this project is to understand how force generation by kinesin-related motors and dynamic microtubules controls the fidelity of chromosome segregation. Both microtubules and motors represent excellent targets for anti-cancer drugs but to make wise choices for developing therapeutics it is necessary to understand their contribution to cell division in detail. MCAK/Kif2C is a MT depolymerizing kinesin that controls MT length within the spindle and supresses chromosome instability (CIN). Using CRISPR/Cas9 engineered cells, rapamycin-dependent relocalization and long-term live imaging we have pinpointed the precise contribution that this protein makes to enhance the fidelity of chromosome segregation which will enable us to understand why this protein rescues CIN in tumorigenic cells and its future potential as a therapeutic target. MCAK/Kif2C is also strongly associated with centrosomes where it has the potential to suppress MT outgrowth and influence centrosome separation. We have identified two other poorly studied centrosome-associated kinesins, Kif25 and Kif9, that function in centrosome cohesion and centrosome satellite positioning respectively and that also impact centrosome separation and positioning. Altered centrosome positioning in cell culture has limited effects on cell viability. However, Both Kif9 and Kif25 are widely expressed in vertebrates. Because Kif25, in particular, is most highly expressed in brain it affords an opportunity to investigate the role of centrosome cohesion in vertebrate development and neural stem cell division using morpholinos directed against Kif25 introduced into early zebrafish embryos. We aim to use live and fixed-cell imaging tools and transgenic cell lines to investigate, with precision, the contribution that these kinesins make to preserving the fidelity of the genome and to long-term cell fate. Ultimately this knowledge base can be leveraged to develop new therapies that target kinesin enzymology or MT dynamics.

修改的项目摘要/摘要部分该项目的广泛目标是了解与动力蛋白相关的电动机和动态微管产生的力如何控制染色体隔离的保真度。微管和电动机都代表了抗癌药物的出色靶标，但要做出明智的选择来开发治疗剂，有必要详细了解它们对细胞分裂的贡献。 MCAK/KIF2C是一种MT解聚驱动蛋白，可控制纺锤体内的MT长度，并减少染色体不稳定性（CIN）。使用CRISPR/CAS9工程细胞，雷帕霉素依赖性重新定位和长期实时成像，我们确定了该蛋白质为增强染色体隔离的忠诚度所做出的确切贡献，这将使我们能够理解该蛋白质在肿瘤细胞中的CIN及其未来潜力作为治疗目标。 MCAK/KIF2C也与中心体密切相关，其中它有可能抑制MT的生长并影响中心体分离。我们已经确定了另外两个研究的不良中心体相关的驱动蛋白KIF25和KIF9，它们分别在中心体凝聚力和中心体卫星定位中起作用，这也影响了中心体的分离和定位。细胞培养中的中心体位置改变对细胞活力的影响有限。但是，KIF9和KIF25均在脊椎动物中广泛表达。由于KIF25在大脑中尤其是最高表达的，因此，使用针对Zebrafish早期胚胎引入的KIF25的吗啡，可以利用用于脊椎动物发育和神经干细胞分裂的中心体凝聚力在脊椎动物发育和神经干细胞分裂中的作用。我们的目标是使用实时和固定细胞成像工具和转基因细胞系，以精确地研究这些驱动剂在保留基因组和长期细胞命运方面所做的贡献。最终，可以利用这种知识库来开发针对动力蛋白酶学或MT动态的新疗法。