Targeting proteoglycan-mediated signaling in Ewing sarcoma

尤文肉瘤中靶向蛋白多糖介导的信号传导

• 批准号: 10591979
• 负责人: Elena Vasileva
• 金额: $ 11.79万
• 依托单位: CHILDREN'S HOSPITAL OF LOS ANGELES
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-07-01 至 2025-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10591979
• 关键词: Address; Advisory Committees; Affect; Apoptosis; Appearance; Bioinformatics; Bone Tissue; CRISPR/Cas technology; Cancer Biology; Cell Differentiation process; Cell Line; Cell Lineage; Cell Proliferation; Cell Survival; Cells; Communication; Communities; Complex; Data; Data Analyses; Dedications; Development; Developmental Biology; Developmental Cell Biology; Dimethyl Sulfoxide; Disease; EWSR1 gene; Effectiveness; Enzymes; Ewings sarcoma; FLI1 gene; Fibroblast Growth Factor Receptors; Fishes; Foundations; Genes; Genetic; Genetic Models; Genomics; Goals; Heparan Sulfate Proteoglycan; Heparanase inhibitors; Home; Human; Image; In Situ Hybridization; In Vitro; Institution; Knock-out; Label; MAPK3 gene; Malignant - descriptor; Malignant Childhood Neoplasm; Malignant Neoplasms; Mammals; Mediating; Mentors; Mentorship; Metabolism; Modeling; Mus; Neoplasm Metastasis; Neural Crest; Normal Cell; Normal tissue morphology; Pathology; Pharmaceutical Preparations; Phase; Population; Positioning Attribute; Predisposition; Proliferating; Proteoglycan; Proteomics; RNA; RNA marker; Recurrent disease; Reporter; Research; Resolution; Resources; Role; Scheme; Signal Transduction; Soft Tissue Neoplasms; Testing; Tissues; Training; Tumorigenicity; Validation; Work; Xenograft Model; Xenograft procedure; Zebrafish; antagonist; behavioral study; biomarker identification; cancer cell; career; cell motility; cell transformation; cell type; comparative; epigenomics; experience; genome editing; high resolution imaging; human disease; imaging approach; in vivo; in vivo Model; inducible Cre; innovation; insight; molecular targeted therapies; mosaic; mouse model; neoplastic cell; novel strategies; overexpression; patient prognosis; professor; small molecule; stem cell biology; tenure track; transcriptomics; tumor; tumor growth; tumor initiation; tumor microenvironment; tumor progression; tumorigenesis; zebrafish genome

Project Summary/Abstract: Ewing sarcoma (ES) is a malignant bone and soft-tissue tumor with extremely poor prognosis for patients with metastatic or relapsed disease. In the absence of representative genetic models, the developmental cell of origin as well as the mechanisms of tumor initiation remain poorly understood. To address those questions, I have developed an innovative zebrafish model of ES by introducing the human EWSR1-FLI1 oncofusion to the zebrafish genome. This model allows studying the behavior of GFP-labeled cancer cells during tumor initiation and progression in a complex developmental context which is not currently possible in mammals. The fish develop tumors positive for known ES markers, recapitulating the main aspects of human disease. Preliminary data reveal dysregulation of heparan sulfate proteoglycan (HSPG) metabolism and associated activation of ERK signaling in ES cells. Targeting HSPGs with the specific antagonist surfen reduces ERK1/2 signaling and decreases tumorigenicity in vitro and in vivo. My preliminary data suggest that dysregulated HSPG turnover can affect normal cell differentiation leading to cell transformation. I hypothesize that such HSPG- mediated dysregulation of signaling between cancer and normal cells can facilitate ES progression. The aims outlined in this proposal will take advantage of the genetic model of ES in combination with innovative single-cell transcriptomic, genetic, and high-resolution imaging approaches to characterize the type of cells giving rise to ES (Aim 1), identify key effectors in cancer and cancer niche cells promoting the cancer progression (Aim 2), and to target HSPG mediated signaling in genetic zebrafish and xenograft mouse models for ES treatment (Aim 3). Aim1 will be completed in the K99 phase and will be co-mentored by Dr. Crump. Aims 2 and 3 will be initiated during the K99 phase under the mentorship of Dr. Amatruda and my advisory committee and then completed during the R00 phase. Completion of these aims will inform what type of cells contribute to ES development. These findings will determine the role of HSPG in tumor initiation and progression building the foundation for a new strategy of targeting the enzymes involved in HSPG metabolism for ES treatment. The proposed project will lay the groundwork for my career goal of obtaining a position as a tenure-track Assistant Professor at a top-tier academic research institution. During the K99 phase, I will receive mentorship in zebrafish and cancer biology from Dr. Amatruda and training in developmental biology from my co-mentor Dr. Crump. Regular interactions with my other advisory committee will allow me to acquire new expertise in single- cell genomics and tumor microenvironment (Dr. Asgharzadeh), ES pathology (Dr. Triche), and data analysis (Dr. Gai). Both CHLA and USC will provide extensive resources for my career and professional development. As CHLA hosts one of the most experienced communities of pediatric cancer biologists, and USC is a home for experts in developmental and stem cell biology there are few better places to conduct this research dedicated to developmental aspects of ES development to acquire the training to achieve my career goals.

项目摘要/摘要：Ewing肉瘤（ES）是一种恶性骨和软组织肿瘤 转移或复发性疾病患者的预后。在没有代表性遗传模型的情况下 起源的发育细胞以及肿瘤起始的机制仍然很少了解。解决 这些问题，我通过介绍人类EWSR1-FLI1开发了ES的创新斑马鱼模型 对斑马鱼基因组的融合。该模型允许研究GFP标记的癌细胞的行为 在哺乳动物中目前无法使用的复杂发育环境中肿瘤的启动和进展。 鱼会形成已知ES标记阳性的肿瘤，从而概括了人类疾病的主要方面。 初步数据揭示了硫酸乙酰肝素蛋白聚糖（HSPG）代谢的失调和相关的 ES细胞中ERK信号的激活。用特定的拮抗剂靶向HSPG降低ERK1/2 信号传导并降低体外和体内肿瘤性。我的初步数据表明HSPG失调 营业额会影响正常的细胞分化，导致细胞转化。我假设这样的hspg- 癌症和正常细胞之间信号传导失调的介导失调可以促进ES进展。 该提案中概述的目的将利用ES的遗传模型与 创新的单细胞转录组，遗传和高分辨率成像方法以表征该类型 产生ES的细胞（AIM 1），鉴定癌症和癌细胞细胞中促进癌症的关键效应子 进展（AIM 2），并靶向遗传斑马鱼和异种移植小鼠模型中HSPG介导的信号传导 用于ES治疗（AIM 3）。 AIM1将在K99阶段完成，并将由Crump博士共同授予。目标 2和3将在Amatruda博士的指导下在K99阶段启动，我的咨询委员会的指导 然后在R00阶段完成。这些目标的完成将告知哪种类型的单元格有助于 ES发展。这些发现将确定HSPG在肿瘤开始和发展中的作用 针对针对HSPG代谢进行ES治疗的酶的新策略的基础。 拟议的项目将为我的职业目标奠定基础，即获得任期的职位 顶级学术研究机构的助理教授。在K99阶段，我将获得指导 Amatruda博士的斑马鱼和癌症生物学以及我的同事博士的发育生物学培训 压碎。与其他咨询委员会的定期互动将使我能够获得单一的新专业知识 细胞基因组学和肿瘤微环境（Asgharzadeh博士），ES病理学（Driche博士）和数据分析（博士 盖）。 CHLA和USC都将为我的职业和专业发展提供广泛的资源。作为 CHLA是小儿癌症生物学家经验最丰富的社区之一，USC是 发育和干细胞生物学方面的专家几乎没有更好的地方进行这项研究。 ES发展的发展方面以获得培训以实现我的职业目标。