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.

尤文肉瘤（Ewing sarcoma，ES）是一种恶性骨和软组织肿瘤， 转移性或复发性疾病患者的预后。在缺乏代表性遗传模型的情况下， 对起源的发育细胞以及肿瘤起始的机制仍然知之甚少。解决 针对这些问题，我通过引入人类EWSR 1-FLI 1，开发了一种创新的斑马鱼ES模型 与斑马鱼基因组的肿瘤融合。该模型允许研究GFP标记的癌细胞在肿瘤生长过程中的行为。 在复杂的发育环境中的肿瘤起始和进展，这在哺乳动物中目前是不可能的。 这些鱼产生的肿瘤对已知的ES标记呈阳性，重现了人类疾病的主要方面。 初步数据显示，硫酸乙酰肝素蛋白聚糖（HSPG）代谢失调， ES细胞中ERK信号的激活。用特异性拮抗剂surfen靶向HSPGs降低ERK 1/2 信号传导并降低体外和体内的致瘤性。我的初步数据表明HSPG失调 更替可以影响正常细胞分化，导致细胞转化。我假设这种HSPG- 癌症和正常细胞之间的信号传导介导的失调可促进ES进展。 本提案中概述的目标将利用ES的遗传模型， 创新的单细胞转录组学，遗传学和高分辨率成像方法来表征类型 的细胞产生ES（目的1），确定在癌症和癌症小生境细胞促进癌症的关键效应 进展（目的2），并在遗传斑马鱼和异种移植小鼠模型中靶向HSPG介导的信号传导 用于ES治疗（目的3）。Aim 1将在K99阶段完成，并将由Crump博士共同指导。旨在 2和3将在Amatruda博士和我的顾问委员会的指导下，在K99阶段启动 然后在R 00阶段完成。这些目标的完成将告知什么类型的细胞有助于 ES开发。这些发现将确定HSPG在肿瘤发生和发展中的作用， 为ES治疗靶向参与HSPG代谢的酶的新策略奠定了基础。 这个项目将为我的职业目标--获得终身教职--打下基础 顶级学术研究机构的助理教授。在K99阶段，我将接受指导 在斑马鱼和癌症生物学从Amatruda博士和培训在发育生物学从我的共同导师博士。 克兰普定期与我的其他咨询委员会的互动将使我获得新的专业知识，在单一的- 细胞基因组学和肿瘤微环境（Asgharzadeh博士），ES病理学（Triche博士）和数据分析（Dr. Gai）。CHLA和南加州大学将为我的职业生涯和专业发展提供广泛的资源。作为 CHLA拥有最有经验的儿科癌症生物学家社区之一，南加州大学是一个家庭， 在发育和干细胞生物学的专家，很少有更好的地方进行这项研究，致力于 ES开发的发展方面，以获得培训，实现我的职业目标。