Targeting insulin receptor splicing for treatment of rhabdomyosarcoma

靶向胰岛素受体剪接治疗横纹肌肉瘤

基本信息

批准号：
9100118
负责人：
Dawn S Chandler
金额：
$ 19.6万
依托单位：
RESEARCH INST NATIONWIDE CHILDREN'S HOSP
依托单位国家：
美国
项目类别：
财政年份：
2016
资助国家：
美国
起止时间：
2016-06-01 至 2018-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9100118
关键词：
Affect Affinity Alternative Splicing Angiogenic Factor Antibodies Antisense Oligonucleotides Beryllium Binding Binding Proteins Binding Sites Biochemical Biological Assay Blood Vessels Cell Line Cell Proliferation Cell physiology Cells Cellular Assay Characteristics Childhood Rhabdomyosarcoma Chronic Clinical Clinical Trials Collaborations DNA Damage Data Development Drug Targeting Elements Enhancers Environment Exhibits Figs - dietary Funding Gene Expression Generations Goals Grant Growth High-Throughput Nucleotide Sequencing Human Hypoxia IGF1R gene IGF2 gene In Vitro Insulin Receptor Insulin-Like Growth Factor II Insulin-Like-Growth Factor I Receptor Intervention Lead Length Link Malignant Neoplasms Messenger RNA Modeling Mus Pathway interactions Patients Pharmacologic Substance Phenotype Play Predisposition Process Production Protein Isoforms Proteins Proteomics Publishing RNA Binding RNA Sequences RNA Splicing RNA, Messenger, Splicing RNA-Binding Proteins Receptor Gene Regulation Regulatory Element Research Resistance Rhabdomyosarcoma Role Signal Transduction Solid Neoplasm Somatomedins Spliced Genes Staging Stimulus Survival Rate System Technology Testing Therapeutic Therapeutic Intervention Transcript Treatment Protocols Tumor Angiogenesis Variant Vascular Endothelial Cell Vascular Endothelial Growth Factors Work Xenograft Model Xenograft procedure angiogenesis autocrine base cancer cell cancer therapy cell behavior childhood sarcoma design epithelial to mesenchymal transition improved insight mortality neoplastic neoplastic cell novel paracrine public health relevance receptor response sarcoma screening therapy development therapy resistant tumor tumor growth tumor progression tumor xenograft tumorigenesis tumorigenic

项目摘要

DESCRIPTION (provided by applicant): Alternative pre-messenger RNA (pre-mRNA) splicing has emerged as a predominant mechanism for proteomic diversity and gene expression. This diversity has been emphasized by recent work that elucidates a network of alternatively spliced genes in response to cell stimuli that are part of the tumorigenic process (DNA damage and Epithelial to Mesenchymal Transition, EMT, for example). Though the importance of alternative splicing in a number of cell processes has been recognized since its discovery, the mechanisms underlying its regulation remain poorly understood. Solid tumors are characterized by both transient and chronic hypoxia, and the ability for tumor cells to adapt to hypoxia is essential for tumor progression. We have shown recently that stimulation of vascular endothelial cell proliferation and angiogenesis induced by the major angiogenic factor, VEGF, is dependent upon insulin-like growth factor (IGF) signaling. Additionally, both human vascular endothelial cells and cells derived from pediatric sarcomas express predominantly the alternatively spliced form of the Insulin Receptor gene (IN-R), that has high affinity for IGF-2. Importantly, IN-R is spliced in response to hypoxia, and pediatric sarcoma cells secrete IGF-2. Consequently, IN-R splicing appears critical in progression of pediatric sarcomas both from acting as a receptor for autocrine growth of tumor cells, and for paracrine growth of vascular cells, and hence angiogenesis. The primary goal of this proposal is to understand the mechanisms and consequences of IN-R pre-mRNA splicing in response to hypoxia. This work will test the hypothesis that regulatory elements and splicing factors are modulated in response to hypoxia and are involved in the alternative splicing of IN-R to contribute to tumorigenesis. In order to determine the effectors of IN-R hypoxia- induced splicing, we will be screening hypoxia induced splicing using a cellular assay which robustly recapitulates changes in IN-R splicing that are seen in cancer cells. We will utilize the hypoxia-induced splicing assay to identify RNA sequences and their respective binding partners that are necessary for regulation of IN-R pre-mRNA splicing. Furthermore, we will use established mouse xenograft assays and novel antisense oligonucleotides (ASOs) to modulate IN-R splicing and determine the role of the spliced isoforms in tumor progression and as novel targets for therapeutic intervention.

描述（由申请人提供）：选择性前信使 RNA (pre-mRNA) 剪接已成为蛋白质组多样性和基因表达的主要机制，最近阐明了选择性剪接基因网络的工作强调了这种多样性。对细胞刺激的反应是致瘤过程的一部分（例如 DNA 损伤和上皮间质转化，EMT）。自发现以来，细胞过程已得到认可，但其调节机制仍知之甚少。实体瘤的特点是短暂和慢性缺氧，而肿瘤细胞适应缺氧的能力至关重要。肿瘤进展。我们最近表明，主要血管生成因子 VEGF 诱导的血管内皮细胞增殖和血管生成的刺激依赖于胰岛素样生长因子 (IGF) 信号传导。此外，人类血管内皮细胞和源自儿科的细胞。肉瘤主要表达胰岛素受体基因 (IN-R) 的选择性剪接形式，该基因对 IGF-2 具有高亲和力，重要的是，IN-R 的剪接响应于胰岛素受体基因 (IN-R)。经测试，在缺氧和小儿肉瘤细胞分泌 IGF-2 的情况下，IN-R 剪接在小儿肉瘤的进展中似乎至关重要，既可以作为肿瘤细胞自分泌生长的受体，也可以作为血管细胞旁分泌生长的受体，从而促进初级血管生成。本提案的目标是了解 IN-R mRNA 前体剪接响应缺氧的机制和后果。这项工作将检验调节元件和剪接因子因响应而受到调节的假设。缺氧并参与IN-R的选择性剪接以促进肿瘤发生为了确定IN-R缺氧诱导的剪接的效应器，我们将使用细胞测定来筛选缺氧诱导的剪接，该细胞测定可靠地概括了IN的变化。在癌细胞中观察到的 -R 剪接我们将利用缺氧诱导的剪接测定来鉴定调节 IN-R 前体 mRNA 所必需的 RNA 序列及其各自的结合配偶体。此外，我们将使用已建立的小鼠异种移植测定和新型反义寡核苷酸测定（ASO）来调节IN-R剪接并确定剪接亚型在肿瘤进展中的作用并作为治疗干预的新靶标。