Targeting AVIL in Glioblastoma

靶向 AVIL 治疗胶质母细胞瘤

基本信息

批准号：
10554307
负责人：
HUI LI
金额：
$ 48.66万
依托单位：
UNIVERSITY OF VIRGINIA
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-03-01 至 2025-02-28
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10554307
关键词：
AVIL gene Actins Adult Affect Animal Model Animals Astrocytes Automobile Driving Avil Binding Biochemical Biological Assay Biological Markers Biophysics CDK4 gene Cell Culture System Cell Culture Techniques Cell Line Cell Proliferation Cell Survival Cells Childhood Rhabdomyosarcoma Clinical Complex Contact Inhibition Data Development Disease Event F-Actin FOXM1 gene Fibroblasts Gene Fusion Glioblastoma Glioma Goals Human In Vitro Investigation Knock-out Knockout Mice MDM2 gene Malignant - descriptor Malignant Childhood Neoplasm Malignant Neoplasms Malignant neoplasm of brain Measures Mediating Modeling Mus Mutation Oncogenes Oncogenic Operative Surgical Procedures Patients Pharmaceutical Preparations Phenocopy Point Mutation Predisposition Primary Brain Neoplasms Prognosis Proteins Radiation Resistance Role Safety Sampling Small Interfering RNA Source Testing Tetanus Helper Peptide Treatment Efficacy Xenograft Model Xenograft procedure aggressive therapy angiogenesis anti-tumor immune response brain tissue cancer type cell motility cohort efficacy evaluation efficacy testing follow-up in vivo mouse model multimodality neoplastic cell nerve stem cell new therapeutic target novel novel strategies novel therapeutic intervention oncogene addiction overexpression patient derived xenograft model pharmacodynamic biomarker rational design resistance mechanism screening small hairpin RNA small molecule small molecule inhibitor stem stem cells targeted cancer therapy targeted treatment therapeutic biomarker therapeutic target therapeutically effective transcriptome treatment planning tumor tumor microenvironment tumorigenesis

项目摘要

Glioblastoma multiforme (GBM), is the most common and most aggressive malignant primary brain tumor in humans. Despite multimodal aggressive therapy, GBM is fatal—with survival over 3 years being considered long-term. More therapeutic targets and treatment options are clearly needed. In our preliminary study, we identified a novel oncogene, AVIL in GBMs. AVIL gene is overexpressed in all the glioblastoma cell lines, including glioblastoma stem cells, and clinical cases, but is hardly detectable in non-cancer astrocytes, neural stem cells, and brain tissues. Silencing AVIL resulted in complete eradication in GBM cell cultures, but had little effect on the astrocyte control cells. In animal models, silencing AVIL dramatically inhibited in vivo xenografts in mice. Conversely, overexpressing AVIL in cell culture systems promoted tumorigenesis, enabled fibroblasts to escape contact inhibition, and transformed immortalized astrocytes. In patient cohorts, higher expression levels of AVIL in gliomas correlated with worse prognosis. Most importantly, GBM cells are more susceptible to small molecule inhibitors of AVIL. These evidences support the premise that AVIL is an Achilles heel of GBMs, targeting which may be an effective approach for GBMs. In this application, we propose the systematic investigation of AVIL as a novel target against GBMs. Aim 1: Determine the efficacy of targeting AVIL in vitro and test AVIL expression as a biomarker of sensitivity. We have shown the critical role of AVIL in some GBM and GSC cell lines. Here, we will investigate its role in additional glioblastoma lines, GSCs, and short-term explants of GBM patient derived xenograft cultures and potential synergistic effect with CDK4/MDM2. We will test the efficacy of tet-inducible shRNAs targeting AVIL, and small molecule compounds we identified through small molecule screen. In addition, we will determine whether AVIL expression serves as a biomarker for sensitivity to AVIL inhibition. Aim 2: Determine the efficacy of targeting AVIL in animal GBM models. We will use shRNA and small-molecule inhibitors in xenograft models to test both the efficacy and safety of targeting AVIL in vivo. In addition, we will use Avil knockout mouse model by crossing them with glioma mouse models to test whether Avil expression is necessary for glioma tumorigenesis in mouse. Aim 3: Investigate the downstream targets and mechanisms of AVIL inhibition and identify potential pharmacodynamic biomarkers. We have preliminary data suggesting AVIL regulates LIN28B, and FOXM1 may mediate the effect in U87 cells. We will investigate whether LIN28B and FOXM1 are functionally critical targets of AVIL, and whether they are potential pharmacodynamic biomarkers to evaluate treatment efficacy of AVIL-targeting therapy. Aim 4: Investigate potential resistance mechanisms of targeting AVIL. The goal is to anticipate challenges to AVIL-based treatment before they arise in the clinical setting. The proposed study will have a significant impact on the understanding and treatment of GBMs. The findings will pave ways to target AVIL as a novel oncogene, and lead to the development of novel therapeutic approaches for the deadly disease.

多形性胶质母细胞瘤（GBM）是最常见、最具侵袭性的恶性原发性脑肿瘤尽管采取了多种积极治疗，GBM 仍然是致命的——生存期超过 3 年。在我们的初步研究中，我们显然需要更多的治疗目标和治疗方案。鉴定出一种新的致癌基因，GBM 中的 AVIL 基因在所有胶质母细胞瘤细胞系中过度表达，包括胶质母细胞瘤干细胞和临床病例，但在非癌症星形胶质细胞、神经胶质细胞中几乎检测不到沉默 AVIL 会导致 GBM 细胞培养物中的干细胞和脑组织被完全根除。在动物模型中，沉默 AVIL 可显着抑制星形胶质细胞的体内活性。小鼠体内的异种移植物，在细胞培养系统中过度表达 AVIL 促进了肿瘤发生。成纤维细胞逃避接触抑制，并转化永生化星形胶质细胞。胶质瘤中 AVIL 的表达水平与较差的预后相关，最重要的是，GBM 细胞较多。对 AVIL 小分子抑制剂敏感这些证据支持 AVIL 是致命弱点的前提。 GBM 的后跟，针对 GBM 可能是一种有效的方法。对 AVIL 作为 GBM 的新靶点进行系统研究目标 1：确定靶向效果。体外 AVIL 并测试 AVIL 表达作为敏感性的生物标志物我们已经证明了 AVIL 在体外的关键作用。一些 GBM 和 GSC 细胞系在这里，我们将研究其在其他胶质母细胞瘤系、GSC 和细胞系中的作用。 GBM 患者来源的异种移植培养物的短期外植体及其潜在的协同效应我们将测试针对 AVIL 的 tet 诱导型 shRNA 和小分子化合物的功效。我们通过小分子筛选鉴定了另外，我们将确定AVIL表达是否作为。对 AVIL 抑制敏感的生物标志物目标 2：确定针对动物 GBM 的 AVIL 功效。我们将在异种移植模型中使用 shRNA 和小分子抑制剂来测试功效和效果。体内靶向 AVIL 的安全性此外，我们将通过与它们杂交来使用 Avil 敲除小鼠模型。神经胶质瘤小鼠模型来测试 Avil 表达是否是小鼠神经胶质瘤肿瘤发生所必需的。研究 AVIL 抑制的下游靶点和机制并确定潜在的药效学我们有初步数据表明 AVIL 调节 LIN28B，FOXM1 可能介导该作用。我们将研究 LIN28B 和 FOXM1 是否是 AVIL 的功能关键靶标。它们是否是评估 AVIL 靶向治疗效果的潜在药效生物标志物目标 4：研究针对 AVIL 的潜在耐药机制。基于 AVIL 的治疗在临床环境中出现之前所面临的挑战。这些发现对 GBM 的理解和治疗产生了重大影响，将为 AVIL 的治疗目标铺平道路。一种新的致癌基因，并导致开发出针对这种致命疾病的新治疗方法。