A new mouse model for testing splice-switching therapies in IGF-driven cancers

用于测试 IGF 驱动癌症中剪接转换疗法的新小鼠模型

基本信息

批准号：
10202220
负责人：
Dawn S Chandler
金额：
$ 9.1万
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：
2021
资助国家：
美国
起止时间：
2021-03-16 至 2023-02-28
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10202220
关键词：
Address Adult Affect Affinity Alternative Splicing American Animal Model Antisense Oligonucleotides Attention Binding Biological Assay CRISPR/Cas technology Cancer Etiology Cancer Model Carcinoma Cell Proliferation Cells Centers for Disease Control and Prevention (U.S.)Cessation of life Clinic Data Death Rate Diagnosis Differentiation and Growth Disease Engineering Enhancers Epigenetic Process Exons Gene Proteins Genes Genomics Hallmark Cell Hepatocyte High-Throughput Nucleotide Sequencing Human Hypoxia IGF2 gene INSR gene Incidence Insulin Insulin Receptor Insulin-Like Growth Factor I Insulin-Like Growth Factor II Introns Knock-in Length Liver Malignant Neoplasms Malignant neoplasm of liver Manipulative Therapies Mediating Methodology Modality Modeling Molecular Mus Mutate Nucleotides Oligonucleotides Organ Donor Pathway interactions Patients Pattern Phenotype Point Mutation Positioning Attribute Predisposition Primary Malignant Neoplasm of Liver Primary carcinoma of the liver cells Process Protein Isoforms RNA Splicing Receptor Gene Regimen Research Resistance Sampling Severity of illness Signal Pathway Signal Transduction Spliced Genes Stress Technology Testing The Cancer Genome Atlas Therapeutic Therapeutic Intervention Time Tissues Transcript Transfection Translating United States Variant Woman angiogenesis cancer diagnosis cell motility clinically relevant cohort design disease phenotype dosage embryonic stem cell human disease in vitro testing liver transplantation mRNA Precursor male men mortality mouse model neoplastic cell new therapeutic target novel novel therapeutics nucleic acid-based therapeutics nucleotide metabolism preference receptor statistics targeted treatment therapeutic RNA therapeutic evaluation tool treatment response tumor tumorigenesis tumorigenic

项目摘要

ABSTRACT Hepatocellular Carcinoma (HCC) is predicted to be the sixth most commonly diagnosed cancer and the fourth leading cause of cancer death worldwide. Rates of both incidence and mortality are 2 to 3 times higher among men and thus liver cancer ranks second in terms of deaths for males. In the United States alone, an estimated 42,810 adults (31,762 men and 11,048 women) will be diagnosed with primary liver cancer in 2020. These statistics, combined with the fact that the death rate of liver cancer has increased by 43% in the last decade, necessitates unconventional treatment approaches. Genomic studies have established the landscape of molecular changes in HCC, however, only ~25% of tumors harbor known targetable drivers. On the other hand, recent advances in high throughput sequencing technologies have uncovered a surprising number of alternatively spliced variants associated with tumorigenesis, implicating de-regulated splicing in the tumor phenotype. Hence, we have turned our attention to the alternatively or aberrantly spliced transcripts in the HCC “spliceome” to identify new therapeutic targets. Insulin receptor has uniquely evolved to undergo alternative splicing to produce two isoforms: the full- length INSR-B and exon 11 skipped INSR-A isoform. Data from TCGA liver cancer cohorts as well as our own multiple in-house patient cohorts show that normal liver tissue primarily expresses the insulin receptor B isoform, whereas human HCC patient samples express more INSR-A. INSR-A, in addition to binding to insulin, has abnormally high affinity for IGF2 and accelerates the onset of tumor-cell hallmarks like proliferation and angiogenesis. Our data further show that this conversion of INSR-B to INSR-A takes place in the presence of stress conditions such as hypoxia. These observations are particularly relevant to HCC because 1) Hif1a has been shown to be significantly elevated and associated with worse progression in HCC and 2) IGF2 has been referred to as an epigenetic onco-driver of HCC. We therefore hypothesize that altering the splice pattern of INSR in liver cancer will abrogate the proliferative signaling downstream and impede the tumorigenic process. To achieve therapeutic intervention, we propose to use splice-switching oligonucleotide (SSO) technology to restore the normal INSR splicing pattern in liver cells. In this proposal, we aim to generate a clinically relevant mouse model of HCC that faithfully recapitulates the INSR splicing changes seen in the human condition. The current HCC mouse models do not express INSR alternatively spliced isoforms and thus do not predict responsiveness to therapies targeting the IGF pathway. There is therefore a critical need for new mouse models of HCC that will allow accurate testing of therapeutic modalities.

抽象的肝细胞癌（HCC）被预测为第六通常被诊断出的癌症，并且是全世界癌症死亡的第四大原因。事件和死亡率的比率高2至3倍在男性中，肝癌在男性死亡方面排名第二。仅在美国，估计将在2020年被诊断出42,810名成年人（31,762名男性和11,048名女性）。这些统计数据，加上以下事实：肝癌的死亡率在上次增加了43％十年，必要的非常规治疗方法。基因组研究已经建立了景观然而，HCC分子变化的肿瘤中只有约25％含有已知靶向驱动因素。另一方面手，高通量测序技术的最新进展已经发现了令人惊讶的数量或者，与肿瘤发生相关的剪接变体，隐含在肿瘤中的剪接表型。因此，我们将注意力转向了HCC中的替代或异常剪接的转录本 “剪接”以识别新的治疗靶标。胰岛素受体已独特地演变为经历替代剪接以产生两个同工型：长度INSR-B和外显子11跳过INSR-A同工型。来自TCGA肝癌队列以及我们自己的数据多个内部患者队列表明，正常肝组织一级表达胰岛素受体B同工型，而人类HCC患者样品表达更多的insr-a。除了与胰岛素结合外，insr-a还具有对IGF2的高度亲和力，并加速了肿瘤细胞标志，例如增殖和血管生成。我们的数据进一步表明，在存在的情况下，INSR-B转换为INSR-A发生压力条件（例如缺氧）。这些观察结果与HCC特别相关，因为1）HIF1A具有被证明显着升高，并且与HCC的进展和2）IGF2相关被称为HCC的表观遗传驱动器。因此，我们假设改变了剪接模式肝癌中的inisr将消除下游的增殖信号传导并阻碍致瘤过程。为了实现治疗干预，我们建议使用剪接切换寡核苷酸（SSO）技术来恢复肝细胞中正常的insr剪接模式。在此提案中，我们旨在产生与临床相关的 HCC的小鼠模型忠实地概括了在人类状况下看到的insr剪接变化。这当前的HCC鼠标模型未表达INSR或剪接的同工型，因此无法预测对针对IGF途径的疗法的反应。因此，新鼠标型迫切需要 HCC的精确测试。