Oncogenic Drivers of Rhabdomyosarcoma Cell State, Cancer Stem Cells and Metastasis

横纹肌肉瘤细胞状态、癌症干细胞和转移的致癌驱动因素

基本信息

批准号：
10658091
负责人：
David Michael Langenau
金额：
$ 59.87万
依托单位：
MASSACHUSETTS GENERAL HOSPITAL
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：
2023-05-01 至 2028-04-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10658091
关键词：
Adult Antibodies Applications Grants Automobile Driving Bar Codes Biological CD44 gene Cell Differentiation process Cell Line Cell Lineage Cell division Cells Cessation of life Childhood Rhabdomyosarcoma Childhood Soft Tissue Sarcoma Classification Combined Modality Therapy DNA-Binding Proteins Development Diagnosis Differentiation Therapy Disease Engraftment FOXO1A gene Gene Fusion Genes Goals Growth Heterogeneity Human Human Development Image Immune Longitudinal Studies Malignant Neoplasms Mesenchymal Mesenchyme Molecular Mus Muscle Muscle Cells Mutation NOTCH3 gene Neoplasm Metastasis Nonmetastatic Oncogenic Operative Surgical Procedures Optics Outcome PAX3 gene PAX7 gene Pathway interactions Patients Pharmacotherapy Population Process Radiation Radiation therapy Recording of previous events Refractory Regimen Relapse Reporter Reporting Resistance Resolution Rhabdomyosarcoma Role Skeletal Muscle Survival Rate Testing Time United States Work Xenograft procedure Zebrafish cancer stem cell cell killing cell type cellular imaging chemotherapy design effective therapy epithelial to mesenchymal transition experimental study forkhead protein gene network in vivo malignant muscle neoplasm multiple omics myocyte-specific enhancer-binding-factor 2C neoplastic cell new therapeutic target novel osteogenic patient derived xenograft model potential biomarker progenitor programs self-renewal single cell sequencing single-cell RNA sequencing stem cell fate stem cell model stem cell self renewal stem cells therapeutic target time use tool transcription factor translational impact tumor tumor growth tumor heterogeneity

项目摘要

Rhabdomyosarcoma (RMS) is the most common pediatric soft tissue sarcoma in the United States. Fusion- negative (FN) RMS are the most common subtype and are driven by RAS-pathway activation. Despite intensive treatment with radiation, chemotherapy, and surgery, a large fraction of patients develop refractory, metastatic, and relapsed RMS that has survival rates of less than 20%. A major hurdle to the design of new and effective treatments for aggressive RMS can be attributed to our limited understanding of the drivers of cancer stem cell (CSC) self-renewal and metastasis. The long-term goal and overall objective of our studies is to identify CSCs and metastatic cells in FN-RMS and then define molecular pathways that can differentiate these cells into non- proliferative, non-migratory cell types or kill them completely. Our central hypothesis is that FN-RMS CSCs drive tumor growth, therapy-resistance and metastasis. We also hypothesize that the genes and pathways promoting the transition of RMS cells into differentiated non-proliferative, non-metastatic cell types can be therapeutically targeted. The rationale and feasibility of our approach comes from our recent discovery of a novel, molecularly- defined FN-RMS CSC that expresses mesenchymal pathway-enriched genes and shares remarkable similarity to a recently discovered bi-potent, muscle mesenchyme progenitor that can make both muscle and osteogenic cells between 9-14 weeks of human development. This FN-RMS CSC is molecularly-distinct from CSCs reported by others in the field, can be isolated using CD44/CD90 antibodies and FACs, and expresses genes associated with epithelial-to-mesenchymal transition (EMT), which is a major driver of metastasis in other cancers. Aim 1 will identify FN-RMS cell heterogeneity and cell types that drive tumor growth and metastasis using single cell sequencing, lineage and cell fate barcode tracing, and mouse xenograft studies. This work will test the hypothesis that CD44+/CD90+ FN-RMS cells define the CSCs and that these cells are largely quiescent under steady state growth conditions, and yet undergo self-renewal divisions following chemo- and radiation-therapy to drive tumor regrowth and metastasis. Aim 2 will quantify human FN-RMS CSC self-renewal and cell state transitions in vivo at single cell resolution using fluorescent cell state reporters, photoconvertible cell lineage tracing tools, and engraftment into optically-clear immune deficient zebrafish. This aim will test the hypothesis that CSCs undergo asymmetric/symmetric self-renewal divisions following therapy to re-create all the functionally diverse cell types in RMS. Aim 3 will identify the molecular mechanisms driving FN-RMS cell states, testing the hypothesis that DNA binding proteins and transcription factors, including NOTCH3 and MEF2C, are dominant oncogenic drivers of RMS cell fate and independently regulate gene networks that promote CSC and/or differentiated muscle cell states. This work will have a positive translational impact by defining new pathways to kill and/or differentiate FN-RMS CSCs and identifying potential biomarkers of therapy resistance based on retention of CSCs after therapy.

横纹肌肉瘤（RMS）是美国最常见的小儿软组织肉瘤。融合- 负 (FN) RMS 是最常见的亚型，由 RAS 通路激活驱动。尽管密集通过放射、化疗和手术治疗，很大一部分患者会出现难治性、转移性、复发性 RMS 的生存率低于 20%。设计新的、有效的技术的一个主要障碍侵袭性 RMS 的治疗可归因于我们对癌症干细胞驱动因素的了解有限（CSC）自我更新和转移。我们研究的长期目标和总体目标是识别 CSC FN-RMS 中的细胞和转移细胞，然后定义可将这些细胞分化为非转移细胞的分子途径增殖的、非迁移的细胞类型或完全杀死它们。我们的中心假设是 FN-RMS CSC 驱动肿瘤生长、治疗抵抗和转移。我们还假设基因和途径促进 RMS 细胞转变为分化的非增殖性、非转移性细胞类型可以进行治疗有针对性。我们方法的基本原理和可行性来自于我们最近发现的一种新颖的分子- 定义的 FN-RMS CSC 表达富含间充质途径的基因并具有显着的相似性最近发现的双能肌肉间质祖细胞，可以使肌肉和成骨人类发育9-14周之间的细胞。该 FN-RMS CSC 在分子上与报道的 CSC 不同可以使用 CD44/CD90 抗体和 FAC 进行分离，并表达相关基因上皮间质转化（EMT），这是其他癌症转移的主要驱动因素。目标1 将使用单细胞识别 FN-RMS 细胞异质性和驱动肿瘤生长和转移的细胞类型测序、谱系和细胞命运条形码追踪以及小鼠异种移植研究。这项工作将测试假设 CD44+/CD90+ FN-RMS 细胞定义了 CSC，并且这些细胞在很大程度上处于静止状态稳态生长条件，但在化疗和放疗后经历自我更新分裂驱动肿瘤再生和转移。目标 2 将量化人类 FN-RMS CSC 自我更新和细胞状态使用荧光细胞状态报告器、光转换细胞谱系以单细胞分辨率进行体内转变追踪工具，以及植入光学透明的免疫缺陷斑马鱼中。该目标将检验假设 CSC 在治疗后经历不对称/对称的自我更新分裂，以重建所有功能 RMS 中的多种细胞类型。目标 3 将确定驱动 FN-RMS 细胞状态的分子机制，测试假设 DNA 结合蛋白和转录因子（包括 NOTCH3 和 MEF2C）占主导地位 RMS 细胞命运的致癌驱动因素并独立调节促进 CSC 和/或的基因网络分化的肌肉细胞状态。这项工作将通过定义新的途径来产生积极的转化影响杀死和/或区分 FN-RMS CSC，并根据治疗后 CSC 的保留。