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

• 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
• 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

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.

项目摘要 转录物的选择性剪接（alternative splicing，AS）是一种重要的转录后过程 用于许多细胞功能，包括增殖、分化和存活。AS的过程是有规律的 RNA结合蛋白（RBP）及其正确的核定位和与剪接体的相互作用 这些成分形成称为核斑点（NS）的可见致密区室。癌症能够 通过靶向参与EMT、侵袭、细胞周期调控和转录的基因劫持和利用AS， 控制已经被描述为影响转移潜能和治疗抗性。最近， 研究表明，50%的乳腺癌含有至少一种RBP的过度表达或扩增， 从而靶向AS的基因转录物以诱导侵袭性特征。此外，一个新的功能 雌激素受体α（ERα）的研究最近揭示了ERα的非经典RNA结合影响后 转录调节和促进细胞适应性响应环境应激。我们的实验室， 沿着其他研究，最近在NF 1和ERα之间建立了一个调节ERα信号传导的关键联系。NF1 是一种肿瘤抑制基因，编码神经纤维蛋白，当突变时， 预后不良的乳腺癌。这些研究激发了研究 NF 1缺乏的乳腺癌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一种协同作用的能力。 ERα转录活性的阻遏物。然而，目前尚不清楚这种神经纤维蛋白-ER α 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 这个规定。我的F99训练阶段（目标1）的总体目标是确定神经纤维蛋白所起的作用 在ERα转录后调节中的作用。具体而言，目的是1.1）确定神经纤维蛋白对ERα的影响 NF 1突变ER+乳腺癌模型中的依赖性转录后调节，以及1.2）确定 选择性雌激素受体调节剂（SERM）影响ERα转录后调控在乳腺癌中的作用 NF 1缺陷细胞。在F99阶段的培训目标将扩大我的分子技术，测序， 癌症生物学的数据分析。在我的K 00阶段（目标2），我将定义撞击核 神经纤维蛋白通过核斑点的形成和功能进行转录后调节。NF1- 缺陷乳腺癌模型将用于确定核斑点的形成、组成和后处理。 转录和基因调控。这项研究金提供的培训和数据将建立 作为一名独立研究人员，我的科学和专业技能基础。