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%，幸存者患有终身治疗相关残疾。这需要开发新的靶向治疗。胰岛素样生长因子1受体（IGF 1 R）小分子抑制剂在体外和体内对NB细胞表现出抗肿瘤作用。 vivo.然而，鉴于IGF 1 R基因中明显缺乏体细胞突变，仍有待确定哪些NB患者子集将从这种方法中受益。我们假设IGF 1 R的表达是由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，这是一种在约50%的NB患者中发现的小的2-nt缺失。使用荧光素酶传感器试验、常用NB细胞系的免疫印迹和使用基因集富集分析的28个原发性神经母细胞瘤的转录组分析，确定了该3 'UTR变体对IGF 1 R表达的积极作用。 此外，rs3833015过表达的基因也强烈富集IGF 1 R/MAPK通路的成员，IGF 1 R/MAPK通路不仅在细胞增殖中，而且在面对化疗时的细胞存活中起着公认的作用。在无偏表达-数量性状基因座（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IGF 1 R mRNA水平降低相关。我们目前的总体假设是，rs3833015和rs374251改变IGF 1信号（分别向上和向下），导致对常规化疗药物和小分子IGF 1 R抑制剂的耐药性改变。我们将在以下两个目标中探讨这一假设。1.确定rs3833015和rs3743251导致IGF 1 R失调的机制，并鉴定与IGF 1 R过度/低表达相关的新SNP。2.研究IGF 1 R变异体对神经母细胞瘤对小分子IGF 1 R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IGF 1 R 3 'UTR基因型与NB患者的存活率相关联，并比较和对比rs3833015和rs374251肿瘤对常规治疗（环磷酰胺和托泊替康）和IGF 1 R抑制剂的反应。这项研究的主要影响是，它将允许对IGF 1 R抑制剂临床试验中招募的患者进行前瞻性分层，并对这些药物产生更强的反应，从而迎来miR药物基因组学时代。