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

基本信息

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
项目类别：
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

项目摘要

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.

概括在胰腺癌中，剪接是癌症发生和进展的关键生物学过程，但很少有研究。研究了剪接调节因子或其剪接靶标在疾病进展中的作用。吞吐量，体内全基因组遗传筛选促进 KRAS 驱动的胰腺导管的新事件在腺癌 (PDAC) 中，我们最近发现了 RNA 剪接因子 RBFOX2 作为潜在的一种独特作用。我们发现 RBFOX2 核丰度显着升高。低分化的人类 PDAC 中的 RBFOX2 缺失会导致高度转移我们发现 RBFOX2 缺失会促进与干细胞相关的外显子跳跃事件。潜力和入侵，包括 ABI1，我们勇敢地面对 RBFOX2 介导的关键调控因子的剪接转换。细胞侵袭和分化程序对其促进 PDAC 进展的作用至关重要。在目标 1 中，我们将研究这些 RBFOX2 介导的剪接事件在 PDAC 进展中的致癌作用。我们将研究由 RBFOX2 选择性剪接的 ABI1ΔEx9 同工型的致癌作用蛋白质组学方法阐明新的蛋白质相互作用和二次修饰有助于 ABI1ΔEx9 在驱动侵袭表型中的功能在目标 2 中，我们将研究“ESC 样”的贡献。使用 i) 剪接，RBFOX2 调节 PDAC 细胞分化和疾病进展的外显子排除事件转换反义寡核苷酸 (AON) 来模拟外显子跳跃亚型并研究其协同驱动离体干细胞样表型和 ii) 可诱导的 cDNA 异构体以了解其作用在目标 3 中，我们将使用 PDX 开发 RBFOX2 剪接特征。使用最先进的剪接分析技术，从快速复发且无进展疾病的患者中提取 PDO 平台（bulk 和 scRNAseq PacBio/isoseq 与 Illumina 短读长测序相结合）并研究其我们期望我们的研究将揭示 RBFOX2- 的生物学相关性。促进胰腺癌进展中的致癌剪接程序并识别新的分子漏洞可以用来改善胰腺癌患者的疾病结果。