Mechanisms of aberrant ribosomal RNA (rRNA) methylation and altered mRNA translation in cancers

癌症中异常核糖体 RNA (rRNA) 甲基化和 mRNA 翻译改变的机制

基本信息

批准号：
10115526
负责人：
Catherine Denicourt
金额：
$ 35.69万
依托单位：
UNIVERSITY OF TEXAS HLTH SCI CTR HOUSTON
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-03-01 至 2024-02-29
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10115526
关键词：
Affect Affinity Binding Biogenesis Bioinformatics Biological Assay Breast Cancer Model Catalytic Domain Cells Code Complex Data Development Disease Environment Frequencies Future Genes Genetic Transcription Genetic Translation Goals Homeostasis Human Impairment Lead Malignant Neoplasms Mass Spectrum Analysis Medical center Messenger RNA Methylation Methyltransferase Molecular Molecular Machines Neoplasm Metastasis Nucleotides Oncogenes Oncogenic Pathway interactions Pattern Peptidyltransferase Play Process Production Proteins Proteome RNA RNA chemical synthesis RNA methylation Recombinant DNA Reporter Resistance Ribosomal RNA Ribosomes Role Shapes Specificity Testing Texas Therapeutic Intervention Transcript Transcription Process Translating Translations breast cancer progression cancer cell fibrillarin hormone therapy insight malignant breast neoplasm medical schools metaplastic cell transformation methylation pattern mouse model novel novel therapeutics overexpression polysome profiling prevent protein complex public health relevance recruit therapeutic development therapeutic target tumor tumor progression tumorigenesis

项目摘要

Project Summary The fidelity of mRNA translation is essential to maintain cellular homeostasis and prevent the production of aberrant proteins that could lead to disease development. Ribosomes are key components of the core translational machinery whose activity can be modulated by diverse types of nucleotide methylation on the ribosomal RNA (rRNA). A growing body of evidence indicates that the pattern of rRNA methylation is altered in cancers, leading to the synthesis of specialized ribosomes with the unique ability to translate mRNAs coding for oncogenic proteins. rRNA methylation has been shown to dynamically regulate ribosome function and influence their efficiency, accuracy and affinity for certain type of mRNAs. Hence, pathways regulating rRNA methylation can selectively drive the translation of a proteome that supports cellular transformation and tumor development. Our overarching goal is to identify pathways and key players involved in regulating the aberrant methylation of rRNA in cancers as they represent unexploited therapeutics targets. We recently uncovered that the oncogene Pelp1 is an important regulator of rRNA methylation in cancer cells. Our experimental approach, supported by preliminary data, is to elucidate the mechanisms by which Pelp1 regulates rRNA methylation and demonstrate the importance of this process for cancer progression. We hypothesize that Pelp1 supports tumorigenesis by modulating ribosomes translational activity through its ability to regulate rRNA methylation. In Aim1 we will determine whether Pelp1 regulates rRNA methylation by recruiting RNA methyltransferases to the nascent rRNA transcript. Because rRNA methylation plays an important role in regulating the translational capacity of ribosomes, we will assess whether overexpression of Pelp1 changes ribosome efficiency, fidelity, and affinity for certain types of mRNAs using translation reporter assays and a polysome profiling-sequencing approach (Aim 2). Pelp1 is overexpressed in 60-80% of breast cancers, where its level of expression directly correlates with tumor grade, metastasis, and endocrine therapy resistance. In Aim 3, we will use orthotopic mouse model of breast cancer to assess whether the ability of Pelp1 to regulate rRNA methylation is required to support tumorigenesis. Our findings suggest that by regulating rRNA methylation, Pelp1 participates in the translational reprograming of cancer cells, thereby promoting the selective translation of oncogenic genes. The successful completion of our proposed study will yield important insight into mechanisms of aberrant mRNA translation and it roles in oncogenesis.

项目概要 mRNA 翻译的保真度对于维持细胞稳态和防止产生可能导致疾病发展的异常蛋白质。核糖体是核心的关键组成部分翻译机制的活性可以通过不同类型的核苷酸甲基化来调节核糖体 RNA (rRNA)。越来越多的证据表明 rRNA 甲基化模式在癌症，导致特殊核糖体的合成，具有翻译编码 mRNA 的独特能力对于致癌蛋白。 rRNA 甲基化已被证明可以动态调节核糖体功能影响其效率、准确性和对某些类型 mRNA 的亲和力。因此，调节 rRNA 的途径甲基化可以选择性地驱动支持细胞转化和肿瘤的蛋白质组的翻译发展。我们的首要目标是确定参与监管异常行为的途径和关键参与者癌症中 rRNA 的甲基化，因为它们代表了未开发的治疗靶点。我们最近发现癌基因 Pelp1 是癌细胞中 rRNA 甲基化的重要调节因子。我们的实验方法，在初步数据的支持下，旨在阐明 Pelp1 调节 rRNA 甲基化和证明这一过程对癌症进展的重要性。我们假设 Pelp1 支持通过核糖体调节 rRNA 甲基化的能力来调节核糖体翻译活性，从而抑制肿瘤发生。在目标1 我们将确定 Pelp1 是否通过向 rRNA 招募 RNA 甲基转移酶来调节 rRNA 甲基化新生rRNA转录本。因为rRNA甲基化在调节翻译过程中起着重要作用核糖体的容量，我们将评估 Pelp1 的过度表达是否会改变核糖体的效率、保真度、使用翻译报告分析和多核糖体分析测序对某些类型 mRNA 的亲和力方法（目标 2）。 Pelp1 在 60-80% 的乳腺癌中过度表达，其表达水平直接影响与肿瘤分级、转移和内分泌治疗耐药性相关。在目标 3 中，我们将使用原位乳腺癌小鼠模型评估是否需要 Pelp1 调节 rRNA 甲基化的能力支持肿瘤发生。我们的研究结果表明，通过调节 rRNA 甲基化，Pelp1 参与癌细胞的翻译重编程，从而促进致癌基因的选择性翻译。这成功完成我们提出的研究将对异常 mRNA 的机制产生重要的见解翻译及其在肿瘤发生中的作用。