Impact of neurofibromin on ER-alpha and nuclear speck post-transcriptional gene regulation

神经纤维蛋白对 ER-α 和核斑点转录后基因调控的影响

基本信息

批准号：
10744501
负责人：
Patrick Dischinger
金额：
$ 4.82万
依托单位：
VAN ANDEL RESEARCH INSTITUTE
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：
2023-09-01 至 2025-08-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10744501
关键词：
Alternative Splicing Award Breast Cancer Model Breast Cancer Patient Breast Cancer cell line Cancer Biology Cell Cycle Regulation Cell Line Cell physiology Cells Characteristics Data Data Analyses Data Set Diagnostic or Prognostic Factor Disease Endocrine Estrogen Receptor alpha Estrogen receptor positive Fellowship Foundations Gene Expression Gene Expression Regulation Genes Genetic Transcription Glioblastoma Goals Human Impairment Invaded Knowledge Laboratories Link Malignant Neoplasms Malignant neoplasm of lung Mammary Neoplasms Mass Spectrum Analysis Modeling Molecular Motivation Mutate NF1 gene Neoplasm Metastasis Nuclear Nuclear Protein Outcome Pathway interactions Phase Play Post-Transcriptional Regulation Postdoctoral Fellow Process Prognosis Proliferating Protein Isoforms Proteins RNA RNA Binding RNA Splicing RNA-Binding Proteins RNA-Protein Interaction Rattus Regulation Research Research Personnel Resistance Role Selective Estrogen Receptor Modulators Signal Transduction Spliced Genes Spliceosomes Techniques Tertiary Protein Structure Therapeutic Therapeutic Intervention Training Transcript Transcription Process Transcriptional Regulation Tumor Suppressor Genes biological adaptation to stress breast cancer progression career crosslink efficacy evaluation environmental stressor epigenomics fitness fluorescence imaging genetic corepressor in vivo inhibitor leukemia loss of function loss of function mutation malignant breast neoplasm mutant next generation sequencing novel novel diagnostics novel therapeutic intervention novel therapeutics overexpression posttranscriptional response skills therapy resistant transcriptomics tumor progression

项目摘要

PROJECT SUMMARY Alternative splicing (AS) of transcripts is an essential post-transcriptional process that is instrumental for numerous cell functions including proliferation, differentiation, and survival. The process of AS is regulated by RNA binding proteins (RBPs) and their proper nuclear localization and interactions with spliceosome components which form visibly dense compartments called nuclear specks (NS). The ability for cancer to hijack and utilize AS by targeting genes involved in EMT, invasion, cell cycle regulation, and transcriptional control has been described to influence metastatic potential and therapeutic resistance. More recently, it has been shown that 50% of breast cancers contain overexpression or amplification of at least one RBP which consequently targets gene transcripts for AS to induce invasive characteristics. Furthermore, a novel function of estrogen receptor alpha (ERα) recently revealed non-canonical RNA binding of ERα to influence post- transcriptional regulation and promote cell fitness in response to environmental stressors. Our laboratory, along with other, recently established a critical link between NF1 and ERα in regulation of ERα signaling. NF1 is a tumor suppressor gene, encoding for the protein neurofibromin, that when mutated drives of a subset of breast cancers with poor prognosis. These studies have sparked motivation to investigate mechanisms in which NF1-deficient breast cancers contribute to metastasis and endocrine resistance. The interaction between neurofibromin and ERα describes neurofibromins ability to act as a co- repressor of ERα’s transcriptional activity. However, it still remains unknown whether this neurofibromin-ERα interaction regulates ERα’s post-transcriptional regulation or the importance of neurofibromin localization for this regulation. The overall goal of my F99 training phase (Aim 1) is to determine the role neurofibromin plays in ERα post-transcriptional regulation. Specifically, the Aims are 1.1) to define neurofibromin’s impact on ERα dependent post-transcriptional regulation in NF1 mutant ER+ breast cancer models and 1.2) determine the efficacy of selective estrogen receptor modulators (SERMs) to impact ERα post-transcriptional regulation in NF1-deficient cells. The training objectives in the F99 phase will expand my molecular techniques, sequencing, and data analysis in cancer biology. During my K00 phase (Aim 2) I will define the impact nuclear neurofibromin has on post-transcriptional regulation through nuclear speck formation and function. NF1- deficient breast cancer models will be used to determine nuclear speck formation, composition, and post- transcriptional and gene regulation. The training and data resulting from this fellowship award will establish the foundation of scientific and professional skills for my career as an independent researcher.

项目概要转录本的选择性剪接 (AS) 是一个重要的转录后过程，具有重要作用 AS 的过程受到调节，包括增殖、分化和存活等多种细胞功能。 RNA 结合蛋白 (RBP) 及其正确的核定位以及与剪接体的相互作用形成明显致密隔室的成分称为核斑点（NS）。通过靶向参与 EMT、侵袭、细胞周期调节和转录的基因来劫持和利用 AS 控制已被描述为影响转移潜力和治疗耐药性。研究表明，50% 的乳腺癌含有至少一种 RBP 的过度表达或扩增，因此，靶向 AS 的基因转录物以诱导侵袭特性。雌激素受体 α (ERα) 的研究最近揭示了 ERα 的非典型 RNA 结合影响后转录调节并促进细胞适应环境压力。与其他人一起，最近在 NF1 和 ERα 之间建立了 ERα 信号传导调节的关键联系。是一种肿瘤抑制基因，编码神经纤维蛋白，当其子集发生突变时这些研究激发了研究预后不良的乳腺癌机制的动力。缺乏 NF1 的乳腺癌会导致转移和内分泌抵抗。神经纤维蛋白和 ERα 之间的相互作用描述了神经纤维蛋白作为协同因子的能力。然而，神经纤维蛋白-ERα是否是ERα转录活性的阻遏物仍然未知。相互作用调节 ERα 的转录后调节或神经纤维蛋白定位的重要性我的 F99 训练阶段（目标 1）的总体目标是确定神经纤维蛋白的作用。具体而言，目标是 1.1) 定义神经纤维蛋白对 ERα 的影响。 NF1 突变 ER+ 乳腺癌模型中的依赖转录后调节和 1.2) 确定选择性雌激素受体调节剂（SERM）影响 ERα 转录后调节的功效 F99 阶段的 NF1 缺陷细胞的训练目标将扩展我的分子技术、测序、在我的 K00 阶段（目标 2），我将定义影响核。神经纤维蛋白通过核斑点形成和功能发挥转录后调节作用。有缺陷的乳腺癌模型将用于确定核斑点的形成、组成和后处理该奖学金所产生的培训和数据将建立转录和基因调控。为我作为独立研究员的职业生涯奠定科学和专业技能的基础。