Molecular Modeling of Pediatric Skeletal Muscle Tumors

儿童骨骼肌肿瘤的分子模型

基本信息

批准号：
8385551
负责人：
Corinne Mary Linardic
金额：
$ 26.56万
依托单位：
DUKE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-01-01 至 2014-11-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8385551
关键词：
Accounting Address Age Alveolar Apoptosis Appearance Behavior Biochemical Biological Assay Bypass Catalytic Domain Cell Line Cell Proliferation Cells Child Childhood Collection Complement Complementary DNA DNA Tumor Viruses Development Disease Disease model Embryonal Rhabdomyosarcoma Enzymes Event FOXO1A gene GTP-Binding Proteins Growth Histologic Human Immunoblotting Immunodeficient Mouse Investigation Knowledge Light Malignant - descriptor Malignant Childhood Neoplasm Malignant Neoplasms Mediating Microscopic Modeling Molecular Models Monitor Muscle Fibers Mutation Myoblasts Names Oncogenes Oncogenic Outcome PAX3 gene Pathway interactions Patients Phenotype Population Proteins Ras Signaling Pathway Receptor Protein-Tyrosine Kinases Research Rhabdomyosarcoma Role Series Signal Transduction Simian virus 40 Skeletal Muscle Skeletal Muscle Neoplasm Solid Neoplasm Telomerase Testing Tumor Suppressor Proteins Up-Regulation Xenograft procedure angiogenesis base cancer cell cell transformation fetal fusion gene gain of function high risk in vivo insight light microscopy loss of function molecular modeling neoplastic cell new therapeutic target overexpression postnatal human research study sarcoma senescence skeletal muscle differentiation soft tissue standard care transgene expression tumor tumor xenograft tumorigenesis tumorigenic vector

项目摘要

ABSTRACT Rhabdomyosarcoma (RMS) is a heterogeneous collection of cancers demonstrating varying degrees of skeletal muscle differentiation. Although accounting for ~8% of pediatric malignant solid tumors, RMS is the most common soft tissue sarcoma in children younger than 14 years. The two major histologic subtypes of RMS are embryonal (eRMS) and alveolar (aRMS). High risk patients have a 5-year survival of 30%, and outcome is very poor for children whose tumors express the PAX3-FKHR fusion gene; when metastatic, their 5-year survival is <8%. This signature genetic change is found only in aRMS and considered a tumor-specific oncogene, but has no molecularly targeted treatment. To address gaps in knowledge of RMS, we have created a new model for this disease based on the conversion of primary human skeletal muscle cells to their tumorigenic counterpart, using a defined set of genetic changes. Using this model, we found that human skeletal muscle myoblasts may be converted to cells that generate tumors mimicking RMS when tested as xenografts in immunodeficient mice. Having established that primary human cells of skeletal muscle origin can give rise to RMS, we studied the repercussions of expressing PAX3-FKHR in them, and discovered two phenotypes that may underlie its oncogenic behavior. First, when PAX3-FKHR was stably expressed as an early genetic change, it enabled bypass of the senescence checkpoint and served as an initiating oncogenic hit for the development of skeletal muscle tumors. Second, when PAX3-FKHR was stably expressed as a late genetic change, it shortened the latency of in vivo tumor formation from 11 to 2 weeks, possibly through activation of the Ras pathway, since in control experiments PAX3-FKHR could functionally substitute for the RAS oncogene. In this proposal, we wish to understand how PAX3-FKHR enables bypass of the senescence checkpoint, and how it accelerates tumorigenesis in previously transformed cells. To accomplish this, we will (1) examine candidate proteins that are downstream of PAX3-FKHR for their role in overcoming the senescence checkpoint, using both gain-of-function and loss-of-function approaches, and (2) examine the accelerated tumor cells for enhanced self-sufficiency in growth signaling, apoptosis, and/or angiogenesis, and the role of the Ras pathway in this PAX3-FKHR-augmented tumorigenesis. The accomplishment of these aims will provide insight into the genesis of this pediatric malignancy, and provide new therapeutic targets for study. In addition, this genetically defined model will serve as a template for the systematic investigation of other human sarcomas.

抽象的横纹肌肉瘤 (RMS) 是一种异质性癌症集合，表现出不同程度的症状骨骼肌的分化。尽管 RMS 约占儿童恶性实体瘤的 8%，但 14 岁以下儿童中最常见的软组织肉瘤。两种主要的组织学亚型 RMS 包括胚胎 (eRMS) 和肺泡 (aRMS)。高风险患者的 5 年生存率为 30%，并且对于肿瘤表达 PAX3-FKHR 融合基因的儿童，预后非常差；当转移时，他们的 5 年生存率<8%。这种标志性基因变化仅在 aRMS 中发现，并被认为是肿瘤特异性的癌基因，但尚无分子靶向治疗。为了解决 RMS 知识方面的差距，我们有基于原始人类骨骼肌细胞向其细胞的转化，创建了这种疾病的新模型致瘤对应物，使用一组定义的遗传变化。利用这个模型，我们发现人类当进行如下测试时，骨骼肌成肌细胞可能会转化为产生模仿 RMS 肿瘤的细胞免疫缺陷小鼠的异种移植。已确定骨骼肌起源的原始人类细胞可以引起 RMS，我们研究了在其中表达 PAX3-FKHR 的影响，并发现了两个可能是其致癌行为的表型。首先，当 PAX3-FKHR 稳定表达为早期基因变化，它能够绕过衰老检查点并充当起始致癌物骨骼肌肿瘤的发展受到打击。其次，当PAX3-FKHR作为晚期稳定表达时基因改变，将体内肿瘤形成潜伏期从 11 周缩短至 2 周，可能是通过 Ras 途径的激活，因为在对照实验中 PAX3-FKHR 可以在功能上替代 RAS癌基因。在本提案中，我们希望了解 PAX3-FKHR 如何绕过衰老检查点，以及它如何加速先前转化细胞的肿瘤发生。为了实现这一目标，我们将 (1) 检查 PAX3-FKHR 下游的候选蛋白在克服衰老检查点，使用功能获得和功能丧失方法，以及（2）检查加速肿瘤细胞增强生长信号、细胞凋亡和/或血管生成的自给自足，以及 Ras 通路在 PAX3-FKHR 增强的肿瘤发生中的作用。这些成绩的实现目标将深入了解这种儿科恶性肿瘤的起源，并为治疗提供新的治疗靶点学习。此外，这个基因定义的模型将作为系统研究的模板其他人类肉瘤。

项目成果

期刊论文数量（2）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Human rhabdomyosarcoma cell lines for rhabdomyosarcoma research: utility and pitfalls.

用于横纹肌肉瘤研究的人类横纹肌肉瘤细胞系：实用性和陷阱。

DOI：
发表时间：
2013
期刊：
影响因子：
0
作者：
Hinson, Ashley R P;Jones, Rosanne;Crose, Lisa E S;Belyea, Brian C;Barr, Frederic G;Linardic, Corinne M
通讯作者：
Linardic, Corinne M

RAS and ROS in rhabdomyosarcoma.

横纹肌肉瘤中的 RAS 和 ROS。