IGF1R gene 3'UTR variants in high-risk pediatric neuroblastoma

高危儿童神经母细胞瘤中 IGF1R 基因 3UTR 变异

• 批准号: 8605178
• 负责人: Andrei Thomas-Tikhonenko
• 金额: $ 21.2万
• 依托单位: CHILDREN'S HOSP OF PHILADELPHIA
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-01-14 至 2015-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8605178
• 关键词: 3' Untranslated Regions; Accounting; Affect; Alleles; Binding; Binding Sites; Biological Assay; Biological Markers; Cell Line; Cell Proliferation; Cell Survival; Cessation of life; Childhood; Childhood Extracranial Solid Tumor; Clinical Trials; Collaborations; Cyclophosphamide; DNA-Binding Proteins; Data; Development; Disease; Enrollment; Event; Exhibits; Future; Gene Expression; Gene Expression Profile; Genes; Genetic Polymorphism; Genome; Genomics; Genotype; Germ-Line Mutation; IGF1 gene; IGF1R gene; Immunoblotting; In Vitro; Insulin; Insulin-Like-Growth Factor I Receptor; Length; Life; Link; Luciferases; MAP Kinase Gene; MYCN gene; Malignant Childhood Neoplasm; Malignant Neoplasms; Messenger RNA; MicroRNAs; Minor; Molecular; Mutation; Neuroblastoma; Nuclear; Nucleotides; Oncogenes; Open Reading Frames; Outcome; Pathway interactions; Patients; Pharmaceutical Preparations; Pharmacogenomics; Phosphotransferases; Play; Population; Positioning Attribute; Quantitative Trait Loci; Receptor Tyrosine Kinase Gene; Reliance; Repression; Resistance; Retroviridae; Role; Signal Transduction; Somatic Mutation; Stratification; Survivors; Therapeutic; Topotecan; Tyrosine Kinase Inhibitor; Untranslated Regions; Validation; Variant; Work; base; chemotherapeutic agent; chemotherapy; cohort; disability; essays; genetic variant; genome-wide; high risk; in vivo; inhibitor/antagonist; innovation; member; mutant; neuroblastoma cell; overexpression; prospective; public health relevance; receptor; receptor expression; research study; response; sensor; small molecule; tumor

DESCRIPTION (provided by applicant): For patients with high-risk neuroblastoma (NB) survival remains below 40%, and survivors suffer from life-long treatment-related disabilities. This necessitates the development of new targeted therapies. Small-molecule inhibitors of insulin-like growth factor 1 receptor (IGF1R) exhibit antitumor effects on NB cells in vitro and in vivo. Yet given the apparent lack of somatic mutations in the IGF1R gene, it remains to be determined what subset of NB patients would benefit from this approach. We hypothesized that IGF1R expression is controlled by alterations in the 3'UTR which would affect binding by microRNAs. In our Preliminary Experiments, we first focused on the genetic variant rs3833015, which is a small 2-nt deletion found in ~50% of NB patients. The positive effects of this 3'UTR variant on IGF1R expression were established using luciferase sensor essays, immunoblotting of commonly available NB cell lines, and transcriptome profiling of 28 primary neuroblastomas using Gene Set Enrichment Analysis. Furthermore, rs3833015-overexpressed genes were also strongly enrichment for members of the IGF1R/MAPK pathway, which plays a well-established role not only in cell proliferation but also in cell survival in the face of chemotherapy. In paralel, during the course of unbiased expression-quantitative trait loci (eQTL) studies, we identified another variant, rs3743251, which creates a new miR-binding site and correlates with decreased IGF1R mRNA levels. Our current overall hypothesis is that rs3833015 and rs374251 change IGF1 signaling (up and down, respectively), resulting in altered resistance to conventional chemotherapeutic agents and small-molecule IGF1R inhibitors. We will pursue this hypothesis in the following two Aims. 1. To determine the mechanisms of IGF1R deregulation by rs3833015 and rs3743251 and to identify new SNPs linked to IGF1R over/under-expression. 2. To investigate the contribution of IGF1R variants to neuroblastoma sensitivity to small-molecule IGF1R inhibitors and chemotherapeutic agents. To this end, we will correlate IGF1R 3'UTR genotypes with survival of NB patients using Kaplan-Meier analysis and compare and contrast responses of rs3833015 and rs374251 tumors to conventional therapeutics (cyclophosphamide and topotecan) and IGF1R inhibitors. The main impact of this study is that it would allow prospective stratification of patients enrolled in clinical trials of IGF1R inhibitors nd more robust responses to these drugs, ushering in the era of miR pharmacogenomics.

描述（由申请人提供）：对于高危神经母细胞瘤 (NB) 患者，生存率仍低于 40%，幸存者患有与治疗相关的终生残疾。这需要开发新的靶向疗法。胰岛素样生长因子 1 受体 (IGF1R) 的小分子抑制剂在体外和体内对 NB 细胞表现出抗肿瘤作用 体内。然而，鉴于 IGF1R 基因明显缺乏体细胞突变，仍有待确定哪些 NB 患者亚群会从这种方法中受益。我们假设 IGF1R 表达是由 3'UTR 的改变控制的，这会影响 microRNA 的结合。在我们的初步实验中，我们首先关注遗传变异 rs3833015，这是在约 50% 的 NB 患者中发现的一个小的 2-nt 缺失。使用荧光素酶传感器文章、常用 NB 细胞系的免疫印迹以及使用基因集富集分析对 28 种原发性神经母细胞瘤进行转录组分析，确定了这种 3'UTR 变体对 IGF1R 表达的积极影响。 此外，rs3833015 过表达基因也强烈富集 IGF1R/MAPK 通路成员，该通路不仅在细胞增殖中发挥着明确的作用，而且在化疗时的细胞存活中也发挥着明确的作用。与此同时，在无偏表达数量性状位点 (eQTL) 研究过程中，我们发现了另一个变体 rs3743251，它创建了一个新的 miR 结合位点，并与 IGF1R mRNA 水平降低相关。我们目前的总体假设是 rs3833015 和 rs374251 改变 IGF1 信号传导（分别上调和下调），导致对传统化疗药物和小分子 IGF1R 抑制剂的耐药性改变。我们将在以下两个目标中追求这一假设。 1. 确定 rs3833015 和 rs3743251 解除 IGF1R 调节的机制，并鉴定与 IGF1R 过表达/表达不足相关的新 SNP。 2. 探讨IGF1R变异对神经母细胞瘤对小分子IGF1R抑制剂和化疗药物敏感性的影响。为此，我们将使用 Kaplan-Meier 分析将 IGF1R 3'UTR 基因型与 NB 患者的生存相关联，并比较和对比 rs3833015 和 rs374251 肿瘤对常规治疗药物（环磷酰胺和托泊替康）和 IGF1R 抑制剂的反应。这项研究的主要影响在于，它将允许对参加 IGF1R 抑制剂临床试验的患者进行前瞻性分层，并对这些药物产生更强烈的反应，从而开创 miR 药物基因组学时代。