Convergent Drivers of Tumor Evolution at the Mitotic Spindle

有丝分裂纺锤体肿瘤进化的趋同驱动因素

基本信息

批准号：
10654843
负责人：
Ruhee Dere
金额：
$ 56.03万
依托单位：
VANDERBILT UNIVERSITY MEDICAL CENTER
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：
2022-07-01 至 2027-06-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10654843
关键词：
Address Alleles Automobile Driving Biochemical Biological Assay Biology Cell division Cells Centrosome Chromatin Chromosome 3 Chromosome abnormality Chromosomes Clear cell renal cell carcinoma Clustered Regularly Interspaced Short Palindromic Repeats Complex Cytoskeleton Data Defect Development Disease Disease Progression Dropout Equilibrium Etiology Evolution Failure Genes Genome Stability Genomic Instability Genomics Heterozygote Histones Human Intervention Kidney Link Loss of Heterozygosity Lysine Malignant Neoplasms Mediating Methylation Methyltransferase Microtubules Mitosis Mitotic Mitotic spindle Modeling Molecular Mutation Oncogenic Organoids Pathway interactions Patients Phenotype Phosphorylation Phosphotransferases Proteins Publishing Reader Regulation Regulatory Element Reporting Role Route Specimen Testing Therapeutic Intervention Time Tubulin Tumor Suppression Tumor Suppressor Proteins Work aurora kinase aurora kinase A carcinogenesis chromatin remodeling chromosome 5q loss combinatorial genome integrity histone methyltransferase innovation insight loss of function member mutant novel novel therapeutic intervention pharmacologic preservation prevent recruit screening targeted treatment therapeutic evaluation tool tumor ubiquitin isopeptidase ubiquitin-protein ligase

项目摘要

Clear cell renal cell carcinoma (ccRCC) is characterized by two major chromosome abnormalities, loss of 3p and gain of 5q, which occur nearly universally in this disease. Here we focus on the unique insights these defects provide into activities at the nexus of chromatin and cytoskeletal biology: coordinated activity of chromatin remodelers on spindle microtubules needed for the integrity of mitosis. Colocalized on chromosome 3p are the chromatin remodelers SETD2, PBRM1, and BAP1 and the E3 ligase VHL. Our groups together have pioneered the concept that SETD2 and PBRM1 have active roles on the cytoskeleton that regulate mitotic fidelity, with impacts on genomic integrity that are only now being revealed. SETD2 is a methyltransferase for both histones and spindle microtubules. Using mutant alleles, we isolated loss of microtubule methylation as underlying the genomic instability tied to SETD2 loss. We further discovered that PBRM1, a substrate recognition member of the PBAF chromatin remodeler, specifically recognizes the SETD2 methyl mark on microtubules, and like the mark it “reads”, is required for genomic stability. Finally, we identified the mitotic kinase AURKA as a new target for VHL-mediated degradation, linking this canonical protein (VHL) to complex regulation of mitotic spindle assembly in ccRCC. In Preliminary Data, we find in addition to VHL loss stabilizing AURKA, AURKA regulates SETD2 via phosphorylation on S2080, connecting VHL and SETD2 for the first time in a common oncogenic pathway. We have also discovered another chromatin remodeler, the chromosome 5q histone methyltransferase NSD1, is also acting at the spindle, and excitingly scores in a CRISPR synthetic lethality screen with SETD2 loss. Our Overarching Hypothesis for this application is that VHL and RCC-associated 3p and 5q chromatin remodelers coordinately regulate methylation of spindle microtubules to maintain genomic stability, which when defective, offers unique opportunities for therapeutic intervention. To address our Overarching Hypothesis, we offer three Specific Aims: We will 1) dissect the convergence on the mitotic spindle of 3p and 5q ccRCC chromatin remodelers, 2) define the features of their interactions at the molecular and biochemical levels that promote mitotic integrity or failure, and 3) mechanistically evaluate points of intervention that lend insight into the controls governing mitotic spindle integrity. To accomplish these aims, we will use innovative tools and strategies that precisely evaluate combinatorial mono- and bi-allelic loss of 3p and 5q genes that occur during progression of ccRCC, as well as a rich pipeline of primary ccRCC organoids. The classic genomic features of ccRCC provide new insights on how cytoskeletal activities of chromatin remodelers converge to maintain genomic stability, the principles of study are broadly applicable to many other cancers and have important ramifications for the development of targeted therapies for this disease.

透明细胞肾细胞癌 (ccRCC) 的特点是两条主要染色体异常，即 3p 缺失和 5q 的增益，这在这种疾病中几乎普遍存在，在这里我们重点关注这些缺陷的独特见解。提供染色质和细胞骨架生物学联系的活动：染色质的协调活动有丝分裂完整性所需的纺锤体微管上的重塑因子共定位于 3p 染色体上。是染色质重塑剂 SETD2、PBRM1 和 BAP1 以及 E3 连接酶 VHL。首创了 SETD2 和 PBRM1 在调节有丝分裂保真度的细胞骨架上发挥积极作用的概念， SETD2 是两者的甲基转移酶，对基因组完整性的影响现在才被揭示。使用突变等位基因，我们将微管甲基化的丧失分离为我们进一步发现 PBRM1（一种底物）与 SETD2 丢失相关。 PBAF染色质重塑者的识别成员，专门识别SETD2甲基标记微管，就像它“读取”的标记一样，是基因组稳定性所必需的。最后，我们确定了有丝分裂。激酶 AURKA 作为 VHL 介导的降解的新靶点，将这种经典蛋白 (VHL) 与复合物联系起来在初步数据中，我们发现除了 VHL 损失稳定外，还有 ccRCC 有丝分裂纺锤体组装的调节。 AURKA，AURKA通过S2080上的磷酸化调节SETD2，首次连接VHL和SETD2 我们还发现了另一种染色质重塑因子，即染色体 5q。组蛋白甲基转移酶 NSD1 也作用于纺锤体，并在 CRISPR 合成中令人兴奋地得分 SETD2 丢失的致死率筛查。我们对此应用的总体假设是 VHL 和 RCC 相关 3p 和 5q 染色质重塑者协调调节纺锤体微管的甲基化以维持基因组稳定性，当有缺陷时，它提供了独特的治疗干预机会来解决我们的问题。总体假设，我们提供三个具体目标：我们将 1) 剖析有丝分裂纺锤体上的收敛 3p 和 5q ccRCC 染色质重塑者，2) 定义了它们在分子和促进有丝分裂完整性或失败的生化水平，以及 3) 机械评估干预点为了深入了解有丝分裂纺锤体完整性的控制，我们将使用。精确评估 3p 和 5q 基因组合单等位基因和双等位基因丢失的创新工具和策略 ccRCC 进展过程中发生的细胞器，以及丰富的原发性 ccRCC 类器官。 ccRCC 的经典基因组特征为染色质的细胞骨架活性如何发挥作用提供了新的见解重塑者聚集以维持基因组稳定性，研究原理广泛适用于许多其他癌症，并对这种疾病的靶向治疗的开发具有重要影响。