RBFOX2 deregulation promotes pancreatic cancer progression through alternative splicing

RBFOX2 失调通过选择性剪接促进胰腺癌进展

• 批准号: 10638347
• 负责人: Karen M Mann
• 金额: $ 46.91万
• 依托单位: H. LEE MOFFITT CANCER CTR & RES INST
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-04-15 至 2028-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10638347
• 关键词: Alternative Splicing; Antisense Oligonucleotides; Automobile Driving; Biological; Biological Process; Cancer Center; Cancer Control; Cell Differentiation process; Cell Line; Cells; Complementary DNA; Coupled; Data; Diagnosis; Differentiation Antigens; Disease; Disease Outcome; Disease Progression; Disseminated Malignant Neoplasm; Down-Regulation; Event; Evolution; Excision; Exclusion; Exhibits; Exons; Flow Cytometry; Genetic Screening; Growth; Human; Immunoprecipitation; Incidence; Intervention; Invaded; KRAS2 gene; Link; Liquid Chromatography; Malignant Neoplasms; Malignant neoplasm of pancreas; Mediating; Metastatic Neoplasm to the Liver; Modeling; Modification; Molecular; Neoplasm Metastasis; Nuclear; Oncogenic; Outcome; Pancreatic Ductal Adenocarcinoma; Pathogenicity; Patient-Focused Outcomes; Patients; Phenotype; Phosphorylation Inhibition; Phosphorylation Site; Play; Process; Progression-Free Survivals; Protein Isoforms; Proteins; Proteomics; RNA Splicing; RNA-Binding Proteins; Recurrence; Recurrent Malignant Neoplasm; Recurrent disease; Regulation; Relapse; Role; Serine; Testing; Threonine; Tissue Microarray; Transcript; Tumor Suppressor Proteins; Tyrosine Phosphorylation; Work; cancer cell differentiation; cancer initiation; cancer recurrence; cancer stem cell; cell growth; cell motility; cohort; crosslinking and immunoprecipitation sequencing; defined contribution; effective therapy; embryonic stem cell; exon skipping; genome-wide; human embryonic stem cell; improved; in vivo; induced pluripotent stem cell; insight; mouse model; novel; pancreatic cancer patients; pancreatic ductal adenocarcinoma cell; pancreatic neoplasm; patient derived xenograft model; prevent; programs; rho; stem cell differentiation; stem cell population; stem cells; stem-like cell; tandem mass spectrometry; transcriptome sequencing; tumor; tumor growth; tumor progression

SUMMARY Splicing is a critical biological process in cancer initiation and progression. In pancreatic cancer, few studies have investigated the role of splicing regulators or their spliced targets in disease progression. Using a high- throughput, genome-wide genetic screen in vivo for novel events promoting KRAS-driven pancreatic ductal adenocarcinoma (PDAC), we recently identified a unique role for the RNA splicing factor RBFOX2 as a latent tumor suppressor driving disease progression. We showed RBFOX2 nuclear abundance is significantly decreased in poorly differentiated human PDAC and that RBFOX2 depletion promotes a highly metastatic disease in orthotopic models. We found RBFOX2 loss promotes exon skipping events associated with stem cell potential and invasion, including ABI1. We hypothesize that RBFOX2-mediated splice-switching in key regulators of cell invasion and differentiation programs is central to its role in promoting PDAC progression. In this proposal, we will examine the oncogenic role of these RBFOX2-mediated splicing events in PDAC progression. In Aim 1, we will investigate the oncogenic role(s) of the ABI1∆Ex9 isoform alternatively spliced by RBFOX2 using proteomics approaches to elucidate novel protein interactions and secondary modifications contributing to ABI1∆Ex9 function in driving invasive phenotypes. In Aim 2, we will investigate the contribution of “ESC-like” exon exclusion events regulated by RBFOX2 to PDAC cell differentiation and disease progression using i) splice- switching Anti-Sense Oligonucleotides (AONs) to mimic exon-skipped isoforms and investigate their cooperativity in driving ex vivo stem cell-like phenotypes and ii) inducible cDNA isoforms to understand their role in promoting disease progression in vivo. In Aim 3, we will develop an RBFOX2 splicing signature using PDXs and PDOs from patients with rapid relapse and progression-free disease using state of the art splicing analysis platforms (bulk and scRNAseq PacBio/isoseq coupled to Illumina short read sequencing) and investigate its association with disease relapse. We expect that our study will reveal the biological relevance of the RBFOX2- promoted oncogenic splicing program in pancreatic cancer progression and identify new molecular vulnerabilities that can be harnessed to improve disease outcomes for pancreatic cancer patients.

概括 剪接是癌症倡议和进展的关键生物学过程。在胰腺癌中，很少有研究 已经研究了剪接调节剂或其剪接靶标在疾病进展中的作用。使用高 用于促进KRAS驱动胰腺导管的新事件的吞吐量，全基因组遗传筛查 腺癌（PDAC），我们最近确定了RNA剪接因子RBFOX2的独特作用 肿瘤抑制疾病进展。我们显示RBFOX2核丰度显着 分化差的人PDAC降低，RBFOX2降低会促进高度转移性 原位模型中的疾病。我们发现RBFOX2损失促进了与干细胞相关的外显子跳过事件 潜力和入侵，包括ABI1。我们假设RBFOX2介导的关键调节器中的剪接切换 细胞侵袭和分化程序的作用是促进PDAC进展中的作用。在此提案中， 我们将研究这些RBFOX2介导的剪接事件在PDAC进程中的致癌作用。在AIM 1中， 我们将研究使用RBFOX2剪接的ABI1ΔEX9同工型的致癌作用 蛋白质组学方法阐明了新型蛋白质相互作用和二次修饰有助于 ABI1ΔEX9在驱动侵入性表型中的功能。在AIM 2中，我们将调查“ ESC般”的贡献 外显子排除事件由RBFOX2调节，用于使用i）剪接 - 使用i） 切换抗沉思寡核苷酸（AONS），以模拟外显子甲板的同工型并研究其 驱动离体干细胞样表型和ii）诱导cDNA同工型的合作性以了解其作用 在体内促进疾病进展。在AIM 3中，我们将使用PDX开发RBFOX2拼接签名 使用最先进的剪接分析的患者和无疾病的患者的PDO 平台（散装和scrnaseq pacbio/isoseq耦合到Illumina简短读取测序）并调查其 与疾病缓解相关。我们预计我们的研究将揭示RBFOX2-的生物学相关性 促进胰腺癌进展的致癌剪接程序并确定新的分子脆弱性 可以利用这可以改善胰腺癌患者的疾病结局。