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) 预计将成为第六大最常诊断的癌症，全球第四大癌症死亡原因，其发病率和死亡率均高出 2 至 3 倍。因此，仅在美国，肝癌就在男性死亡人数中排名第二。预计 2020 年将有 42,810 名成年人（31,762 名男性和 11,048 名女性）被诊断出患有原发性肝癌。这些统计数据，加上去年肝癌死亡率增加了43% 十年来，需要非常规的治疗方法，基因组研究已经确定了这一情况。然而，在 HCC 的分子变化中，只有约 25% 的肿瘤具有已知的靶向驱动因素。另一方面，高通量测序技术的最新进展已经发现了数量惊人的与肿瘤发生相关的选择性剪接变体，暗示肿瘤中剪接失调因此，我们将注意力转向 HCC 中的选择性或异常剪接转录本。 “剪接组”来识别新的治疗靶点。胰岛素受体经过独特的进化，可以进行选择性剪接，产生两种亚型：全亚型长度 INSR-B 和外显子 11 跳过了来自 TCGA 肝癌队列以及我们自己的 INSR-A 亚型数据。多个内部患者队列显示正常肝组织主要表达胰岛素受体 B 亚型，而人类 HCC 患者样本除了与胰岛素结合外，还表达更多的 INSR-A。对 IGF2 异常高的亲和力并加速肿瘤细胞标志的发生，例如增殖和我们的数据进一步表明，INSR-B 向 INSR-A 的这种转化是在存在的情况下发生的。这些观察结果与 HCC 特别相关，因为 1) Hif1a 具有研究显示 IGF2 显着升高并与 HCC 进展恶化相关，并且 2) IGF2 已被证实因此，我们发现它改变了 HCC 的剪接模式。肝癌中的 INSR 将消除下游的增殖信号并阻止致瘤过程。为了实现治疗干预，我们建议使用剪接转换寡核苷酸（SSO）技术恢复肝细胞中正常的 INSR 剪接模式在本提案中，我们的目标是产生临床相关的。 HCC 小鼠模型忠实地再现了人类条件下的 INSR 剪接变化。目前的 HCC 小鼠模型不表达 INSR 选择性剪接异构体，因此无法预测因此，迫切需要新的小鼠模型。 HCC 的检测将允许准确测试治疗方式。