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）是最常见的小儿软组织癌。尽管治疗加剧，晚期RMS患者的结果仍然令人沮丧。截至目前，尚无靶向疗法可用的。显然需要更好的理解和治疗。在我们的初步研究中，我们确定了一个小说癌基因，Avil在RMS中。我们发现，1）Avil在某些RMS中与家具的基因形成融合； 2）在我们测试的大多数RMS中，Avil过表达，但在间充质中几乎无法检测到干细胞或正常肌肉组织； 3）RMS细胞沉迷于Avil失调，因为使火星沉默 Avil（在融合融合的RMS中）或沉默的Avil（在Avil中过表达RMS）几乎消除了细胞，并极大地抑制体内异种移植物，但对对照细胞没有影响。 4）相反，过表达的阿维尔促进了细胞的增殖和迁移，使成纤维细胞形成焦点并转化间充质干细胞； 5）肉瘤表达增加的肉瘤患者的预后较差；和6） RMS细胞对我们抑制阿维尔的小分子敏感。基于这些观察，我们的长期假设是Avil是RMS的致命弱点，而目标可能是治疗治疗的有效方法疾病。在此应用中，我们提出了对Avil作为对RMS的新目标进行系统研究。目的 1：确定靶向Avil在体外的功效。我们将研究其在扩展的单元线列表中的作用和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，和小分子化合物，我们通过小分子筛选鉴定出来。自上次提交以来，我们也产生了70多种初始命中化合物的新型化合物衍生物。我们将在体外测试它们的功效。在此外，我们还将确定阿维尔表达是否是对avil抑制敏感性的生物标志物。目的 2：确定靶向Avil在动物RMS模型中的功效。我们将使用shRNA和小分子异种移植模型中的抑制剂测试靶向Avil在体内的功效和安全性。此外，我们还有同时生成了Avil转基因和敲除模型。我们将以各种CRE菌株将Avil转基因交叉测试Avil过表达是否足以用于RMS肿瘤发生。我们还将使用阿维尔淘汰鼠标通过与RMS鼠标模型跨越模型，以测试RMS是否需要Avil表达小鼠的肿瘤发生。目标3：研究阿维尔抑制的下游目标和机制确定潜在的药效生物标志物。我们将研究分子机制调节FOXM1稳定性，并确定f-actin的域和精确活性致癌活性。 AIM 4：评估靶向AVIL铅化合物的最佳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的理解和治疗的重大影响。这些发现将铺平方法将阿维尔作为一个新颖的癌基因，并导致了绝望疾病的新型治疗方法的发展。