Targeting AVIL, a novel oncogene in rhabdomyosarcoma

针对横纹肌肉瘤中的一种新型癌基因 AVIL

基本信息

批准号：
10585061
负责人：
HUI LI
金额：
$ 49.69万
依托单位：
UNIVERSITY OF VIRGINIA
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-01-01 至 2027-12-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10585061
关键词：
20 year old Adolescent and Young Adult Alveolar Alveolar Rhabdomyosarcoma Animals Automobile Driving Avil Biological Assay Biological Availability Biological Markers Breeding Cell Death Cell Line Cell Proliferation Cells Childhood Collection Combined Modality Therapy Cytoskeleton Data Development Disease Drug Kinetics F-Actin FOXM1 gene Fibroblasts Gene Fusion Gene Knock-Out Model Gene Silencing Genes Genetically Engineered Mouse Goals Histology Housekeeping Gene Housing In Vitro Injections Intravenous Investigation Knockout Mice Knowledge Lead Malignant Childhood Neoplasm Malignant Neoplasms Mesenchymal Stem Cells Metabolism Modeling Molecular Mus Muscle Muscle Development Oncogenes Oncogenic Oral Patient-Focused Outcomes Patients Permeability Pharmaceutical Preparations Phenocopy Planet Mars Predisposition Prognosis Property Proteins RNA RNA Interference Rhabdomyosarcoma Role Safety Series Solubility Testing Tetanus Helper Peptide Tissues Transgenic Mice Transgenic Organisms Xenograft Model Xenograft procedure absorption cell motility chemotherapy efficacy evaluation efficacy testing follow-up in vitro testing in vivo inhibitor mouse model mutant nanomolar new therapeutic target novel novel strategies novel therapeutic intervention overexpression patient derived xenograft model pharmacodynamic biomarker sarcoma screening small hairpin RNA small molecule small molecule inhibitor soft tissue targeted treatment therapeutic biomarker therapeutic target therapeutically effective transcriptome tumorigenesis tumorigenic virtual

项目摘要

Rhabdomyosarcoma (RMS) is the most common pediatric soft-tissue cancer. Despite treatment intensification, the outcome for patients with advanced stage RMS has remained dismal. As of now, no targeted therapy is available. Better understanding and treatment are clearly needed. In our preliminary study, we identified a novel oncogene, AVIL in RMS. We found that, 1) AVIL forms a fusion with a house-keeping gene MARS in some RMS; 2) AVIL is overexpressed in the majority of remaining RMS we tested, yet hardly detectable in mesenchymal stem cells or normal muscle tissues; 3) RMS cells are addicted to AVIL dysregulation in that silencing MARS- AVIL (in RMS cells that harbor the fusion) or silencing AVIL (in AVIL overexpressed RMS) nearly eradicated the cells, and dramatically inhibited in vivo xenografts, but had no effect on control cells; 4) conversely, overexpressing AVIL promoted cell proliferation and migration, enabled fibroblasts to form foci, and transformed mesenchymal stem cells; 5) sarcoma patients with increased AVIL expression have worse prognosis; and 6) RMS cells are sensitive to our small molecules inhibiting AVIL. Based on these observations, our long-term hypothesis is that AVIL is an Achilles heel of RMS, and targeting it may be an effective approach for treating the disease. In this application, we propose the systematic investigation of AVIL as a novel target against RMS. Aim 1: Determine the efficacy of targeting AVIL in vitro. We will investigate its role in an expanded list of cell lines and short-term explants of RMS PDX cultures. We will test the efficacy of tet-inducible shRNAs targeting AVIL, and small molecule compounds we identified through small molecule screen. Since last submission, we also generated over 70 novel compounds derivative of the initial hit compounds. We will test their efficacy in vitro. 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 RMS 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 have generated both Avil transgenic and knockout models. We will cross Avil transgenic with various Cre strains to test whether Avil overexpression is sufficient for RMS tumorigenesis. We will also use Avil knockout mouse model by crossing them with RMS mouse models to test whether Avil expression is necessary for RMS tumorigenesis in mouse. Aim 3: Investigate the downstream targets and mechanisms of AVIL inhibition and identify potential pharmacodynamic biomarkers. We will investigate molecular mechanism under which AVIL regulates FOXM1 stability, and determine the domain and exact activity on F-actin that are responsible for its oncogenic activity. Aim 4: Assess AVIL-targeting lead compounds for optimal ADME (absorption, distribution, metabolism, elimination), bioavailability, and pharmacokinetic properties. The proposed study will have a significant impact on the understanding and treatment of RMS. The findings will pave ways to target AVIL as a novel oncogene, and lead to the development of novel therapeutic approaches for the desperate disease.

横纹肌肉瘤（RMS）是最常见的儿科软组织癌症。尽管加强治疗，晚期 RMS 患者的结果仍然令人沮丧。目前尚无靶向治疗可用的。显然需要更好的理解和治疗。在我们的初步研究中，我们确定了一部小说致癌基因，RMS 中的 AVIL。我们发现，1）AVIL与某些RMS中的看家基因MARS形成融合； 2) AVIL 在我们测试的大多数剩余 RMS 中过度表达，但在间充质中几乎检测不到干细胞或正常肌肉组织； 3) RMS细胞对AVIL失调上瘾，因为MARS- AVIL（在含有融合蛋白的 RMS 细胞中）或沉默 AVIL（在 AVIL 过表达的 RMS 中）几乎消除了细胞，并显着抑制体内异种移植物，但对对照细胞没有影响； 4）相反，过表达 AVIL 促进细胞增殖和迁移，使成纤维细胞形成灶，并转化间充质干细胞； 5) AVIL表达升高的肉瘤患者预后较差；和 6) RMS 细胞对我们抑制 AVIL 的小分子敏感。基于这些观察，我们的长期假设 AVIL 是 RMS 的致命弱点，针对它可能是治疗 RMS 的有效方法疾病。在此应用中，我们建议对 AVIL 作为对抗 RMS 的新靶点进行系统研究。目的 1：确定体外靶向 AVIL 的功效。我们将研究它在扩展的细胞系列表中的作用和 RMS PDX 培养物的短期外植体。我们将测试针对 AVIL 的 tet 诱导型 shRNA 的功效，以及我们通过小分子筛选鉴定出的小分子化合物。自上次提交以来，我们还生成了 70 多种新化合物，这些化合物是最初热门化合物的衍生物。我们将在体外测试它们的功效。在此外，我们将确定 AVIL 表达是否可作为对 AVIL 抑制敏感性的生物标志物。目的 2：确定在动物 RMS 模型中靶向 AVIL 的功效。我们将使用 shRNA 和小分子抑制剂在异种移植模型中测试体内靶向 AVIL 的有效性和安全性。此外，我们还有生成了 Avil 转基因模型和基因敲除模型。我们将 Avil 转基因与各种 Cre 菌株杂交测试 Avil 过度表达是否足以促进 RMS 肿瘤发生。我们还将使用 Avil 基因敲除小鼠通过将它们与 RMS 小鼠模型交叉来测试 Avil 表达是否是 RMS 所必需的模型小鼠肿瘤发生。目标3：研究AVIL抑制和抑制的下游靶点和机制识别潜在的药效生物标志物。我们将研究AVIL的分子机制调节 FOXM1 稳定性，并确定 F-肌动蛋白上负责其稳定性的结构域和确切活性致癌活性。目标 4：评估 AVIL 靶向先导化合物以获得最佳 ADME（吸收、分布、代谢、消除）、生物利用度和药代动力学特性。拟议的研究将有一个对 RMS 的认识和治疗产生重大影响。这些发现将为 AVIL 的目标铺平道路新的致癌基因，并导致针对这种绝望疾病的新治疗方法的开发。