Targeting of RNA-binding protein FXR1 in HNSCC

HNSCC 中 RNA 结合蛋白 FXR1 的靶向

• 批准号: 10571379
• 负责人: Viswanathan Palanisamy
• 金额: $ 41.94万
• 依托单位: UNIVERSITY OF NEW MEXICO HEALTH SCIS CTR
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-08-15 至 2025-08-14
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10571379
• 关键词: Ablation; Affinity; Amino Acids; Animal Model; Arginine; Binding; Biochemical; Biological Assay; Bypass; CDKN1A gene; Cancer Cell Growth; Cell Aging; Cells; Chromosomes; Clinical Trials; Complex; Data; Development; Drug Targeting; Essential Amino Acids; Essential Genes; Event; Evolution; FDA approved; FMR1; G-Quartets; Gene Amplification; Gene Expression; Gene Silencing; Genetic; Glycine; Goals; Growth; Head and Neck Cancer; Head and Neck Squamous Cell Carcinoma; Human; In Vitro; Intervention; Ligands; Lysine; Malignant Neoplasms; Mammalian Cell; Mass Spectrum Analysis; Mediating; Messenger RNA; Methods; Methylation; Methyltransferase; MicroRNAs; Modality; Modeling; Molecular; Mouth Neoplasms; Mutation; Neoplasms; Nuclear Export; Nucleic Acids; Oligonucleotides; Oncogenic; Oncoproteins; Oral; Pathway interactions; Patients; Pharmaceutical Preparations; Play; Post-Transcriptional Regulation; Process; Proliferating; Proteins; Proteomics; Publishing; RNA; RNA Binding; RNA Sequences; RNA Stability; RNA-Binding Proteins; Radiolabeled; Repression; Role; Signal Transduction; Single-Stranded DNA; Specificity; Structure; Survival Rate; TP53 gene; Telomerase RNA Component; Testing; Therapeutic; Translations; Tumor Suppressor Genes; Tumor Suppressor Proteins; Untranslated RNA; Work; Xenograft procedure; aptamer; cancer cell; cancer therapy; cell growth; demethylation; design; diagnostic value; drug resistance development; effective therapy; efficacious treatment; experimental study; improved; in vivo; inhibitor; innovation; insight; mRNA Transcript Degradation; malignant mouth neoplasm; metaplastic cell transformation; mortality; mouth squamous cell carcinoma; mutant; novel; novel strategies; novel therapeutic intervention; novel therapeutics; overexpression; posttranscriptional; pre-clinical; prevent; protein degradation; recruit; small molecule; small molecule inhibitor; telomere; transcriptomics; tumor; tumor growth; tumor progression; tumorigenesis; virtual

Abstract Head and neck squamous cell carcinoma (HNSCC) is the 6th most common cancer worldwide and accounts for more than 90% of head and neck cancers, with a 5-year survival rate of approximately 50%. Despite specific effective and novel therapies, most patients remain unresponsive to current treatment modalities and develop drug resistance. Therefore, identifying new molecules that target cancer pathways, alone or in combination, is critical for developing novel cancer therapies. In HNSCC, dysregulated RNA-binding proteins (RBPs) controls co- and post-transcriptional events. Approaches targeting RBPs are not well established to regulate the gene expression in cancers. Thus, new therapeutic strategies to control gene expression and tumor growth are necessary. Our published findings demonstrate that FXR1 directly binds the RNA G-quadruplex (G4) regions of mRNA CDKN1A and a non-coding RNA TERC (Telomerase RNA component) and promotes the growth and proliferation of HNSCC cells. Herein, our preliminary findings indicate that methylated arginine amino acids at the NES (nuclear export signal) and RGG (arginine-glycine rich) domain of FXR1 are responsible for G4-RNA binding and the growth of cancer cells. However, mutation of specific arginine residues in the NES/RGG domain abolished FXR1 binding with CDKN1A and TERC and reduced the growth and proliferation of oral cancer cells. Thus, targeting FXR1's RGG domain with specific inhibitors is considered an appealing drug target for oral cancer treatment. We plan to use nucleic acid-based aptamers that directly bind to the arginine amino acids present in NES/RGG domain of FXR1 and disrupt its RNA binding activities. Hence, the overall goal of this R21 proposal is to utilize structural and biochemical assays to screen and identify aptamers specific to the FXR1 NES/RGG domain to control its gene expression activities and the growth of oral cancer cells. Two specific aims are proposed to test this model. In specific aim1, we will determine how arginine methylation impacts FXR1 RNA- binding functions and oral tumor progression using in vitro RNA binding assays and in vivo tumor models. In specific aim2, we will develop an aptamer that directly binds and interfere with methylated FXR1-RNA complex and show anti-tumor activity in our preclinical animal models of HNSCC. Our present work is an innovative, straightforward, and robust approach to target the dysregulated RBP-mediated gene expression in HNSCC. We will use these mechanistic findings to develop novel strategies to treat aggressive HNSCC tumors.

摘要 头颈部鳞状细胞癌（HNSCC）是世界上第六大常见癌症，占 超过90%的头颈部癌症，5年生存率约为50%。尽管具体 尽管目前的治疗方法有效且新颖，但大多数患者对目前的治疗方式仍无反应， 耐药性因此，识别靶向癌症通路的新分子，单独或组合， 对于开发新的癌症疗法至关重要。在HNSCC中，调节异常的RNA结合蛋白（RBP）对照 共转录和转录后事件。靶向RBPs的方法还没有很好地建立来调节基因 在癌症中的表达。因此，控制基因表达和肿瘤生长的新的治疗策略正在开发中。 必要我们发表的研究结果表明，FXR 1直接结合RNA G-四链体（G4）区域， mRNA CDKN 1A和非编码RNA TERC（端粒酶RNA组分），并促进生长， HNSCC细胞的增殖。在此，我们的初步研究结果表明，甲基化精氨酸氨基酸在 FXR 1内斯（核输出信号）和RGG（富含甘氨酸的甘氨酸）结构域负责G4-RNA 结合和癌细胞的生长。然而，内斯/RGG结构域中特定精氨酸残基的突变 消除了FXR 1与CDKN 1A和TERC的结合，并降低了口腔癌细胞的生长和增殖。 因此，用特异性抑制剂靶向FXR 1的RGG结构域被认为是口服给药的有吸引力的药物靶标。 癌症治疗我们计划使用基于核酸的适体，直接结合精氨酸氨基酸 存在于FXR 1的内斯/RGG结构域中并破坏其RNA结合活性。因此，R21的总体目标是 一个建议是利用结构和生物化学分析来筛选和鉴定特异于FXR 1的适体 内斯/RGG结构域调控其基因表达活性和口腔癌细胞的生长。两个具体目标 提出了测试这个模型。在特定的aim 1中，我们将确定精氨酸甲基化如何影响FXR 1 RNA- 结合功能和口腔肿瘤的进展，使用体外RNA结合试验和体内肿瘤模型。在 特异性aim 2，我们将开发一种直接结合并干扰甲基化FXR 1-RNA复合物的适体 并在我们的HNSCC临床前动物模型中显示出抗肿瘤活性。我们目前的工作是一个创新的， 这是一种简单、可靠的方法，用于靶向HNSCC中RBP介导的基因表达失调。我们 将利用这些机制的发现，开发新的策略来治疗侵袭性HNSCC肿瘤。