Function and Targeting of ETV6 in Ewing Sarcoma

ETV6 在尤文肉瘤中的功能和靶向

• 批准号: 10740562
• 负责人: Yuan Gao
• 金额: $ 13.58万
• 依托单位: COLD SPRING HARBOR LABORATORY
• 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/10740562
• 关键词: Acute; Advisory Committees; Affect; Allosteric Regulation; Binding; Biochemical; Bone Tissue; Bone neoplasms; CRISPR screen; Cancer cell line; Cancerous; Cell Death; Cell Differentiation process; Cell Line; Cell physiology; Cells; Chemicals; Child; Chimeric Proteins; Chromosomal translocation; Clustered Regularly Interspaced Short Palindromic Repeats; DNA; DNA Binding; DNA Binding Domain; Dedications; Dependence; Development; Differentiated Gene; Differentiation and Growth; Disease; Dominant-Negative Mutation; Drug Targeting; ETV6 gene; EWS-FLI1 fusion protein; Elements; Engineering; Environment; Ewings sarcoma; Exons; FLI1 gene; Faculty; Family; Family member; Funding Mechanisms; Future; Gene Expression; Generations; Genetic study; Genome; Goals; Growth; In Vitro; Knowledge; Maintenance; Malignant Bone Neoplasm; Malignant Neoplasms; Mediating; Mentors; Mesenchymal Differentiation; Metabolic Control; Methodology; Monitor; Mus; Mutagenesis; Mutation; Normal tissue morphology; Oncogenic; Patients; Pediatric Oncology; Penetration; Peptides; Phase; Phenotype; Phosphatidic Acid; Positioning Attribute; Postdoctoral Fellow; Proteins; Reporter; Research; SAM Domain; Site; Soft Tissue Neoplasms; Specific qualifier value; Specificity; Surface; Surveys; System; Testing; Therapeutic Index; Therapeutic Intervention; Tissues; Training; Transcription Repressor; Transcriptional Regulation; Variant; Work; Xenograft Model; anti-cancer; base editing; cancer cell; cancer type; drug development; effective therapy; fitness; functional genomics; in vivo; meetings; mortality; novel; programs; screening; small molecule; structural biology; symposium; targeted treatment; therapeutic target; therapy development; transcription factor; transcriptomics; transition mutation; tumor; tumorigenesis; young adult

PROJECT SUMMARY/ABSTRACT Ewing sarcoma is a highly aggressive bone and soft tissue tumor mainly affecting children and young adults, which still lacks effective therapy. Due to its low mutational burden, there are very few acquired vulnerabilities in Ewing sarcoma cells. During my postdoctoral work, I discovered that transcription factor ETV6 is among the very few top Ewing sarcoma-biased dependencies. Remarkably, although ubiquitously expressed, ETV6 is not essential for cancer cells outside Ewing sarcoma, based on CRISPR screening efforts from us and others. Genetic studies in mice also revealed that ETV6 is dispensable in the majority of tissues. Hence, perturbing ETV6 function in Ewing sarcoma will have a wide therapeutic index. In the proposed project, I aim to further study the detailed mechanisms of how ETV6 maintains the cancer cell state, and leverage this new knowledge to develop therapies for Ewing sarcoma patients with exceptional potency and specificity. To do so, I will characterize the mesenchymal differentiation phenotype of ETV6-deficient Ewing sarcoma cells using high- throughput functional genomics and single cell transcriptomics. Integration of these `-omic' approaches allow me to obtain a deep biochemical understanding of the cancer maintenance function of ETV6 (Aim 1). Moreover, I showed that expression of the Sterile Alpha Motif (SAM) domain of ETV6, which is responsible for its self- oligomerization, has a dominant-negative effect to endogenous ETV6, and inhibits sarcomagenesis in vivo. Therefore, I propose to optimize this SAM peptide to increase its potency and further engineer it for exogenous delivery (Aim 2). Successful generation of a potent, and tumor penetrating ETV6 blocker will benefit therapy development. Finally, as transcription factors, like ETV6, have proven to be challenging targets for drug development, I profiled for endogenous metabolites that can bind ETV6 to unveil druggable pockets, and identified its association with phosphatidic acid. I will further explore the regulatory effects of phosphatidic acid binding to ETV6 in Ewing sarcoma (Aim 3). Results from these studies will guide small molecule development, and more importantly, will also reflect a novel mechanism for metabolic control of gene expression through direct allosteric regulation of transcription factors. During the mentored K99 phase, I will work closely with my mentor Dr. Christopher Vakoc and co-mentor Dr. Carolyn Fein Levy, and collaborators Drs. Stegmaier, Kentsis, Shi and Furukawa, recognized experts in pediatric oncology, peptide therapy, screening methodology and structural biology respectively. I have also established an exceptional advisory committee at CSHL, constituted by Drs. Joshua-Tor and Beyaz, who will monitor and support my transition to independence. In addition, CSHL will provide me an outstanding scientific environment for my research and training, being a conference hub for world-renowned meetings and courses. My objective is to obtain a faculty position to develop an impactful research program, where the K99/R00 funding mechanism will serve as an essential step in my transition to independence.

项目总结/摘要 尤文肉瘤是一种高度侵袭性的骨和软组织肿瘤，主要累及儿童和年轻人， 仍然缺乏有效的治疗方法。由于其低突变负担， 尤文肉瘤细胞。在我的博士后工作中，我发现转录因子ETV 6是 很少有尤因肉瘤的依赖性。值得注意的是，尽管ETV 6广泛表达， 根据我们和其他人的CRISPR筛选工作，它对尤文肉瘤以外的癌细胞至关重要。 在小鼠中的遗传研究也揭示了ETV 6在大多数组织中是可降解的。因此， ETV 6在尤文肉瘤中的功能将具有广泛的治疗指数。在拟议的项目中，我的目标是进一步 研究ETV 6如何维持癌细胞状态的详细机制，并利用这些新知识 为尤文肉瘤患者开发具有特殊效力和特异性的疗法。为此，我将 使用高密度聚乙烯凝胶电泳表征ETV 6缺陷型尤文肉瘤细胞的间充质分化表型， 通量功能基因组学和单细胞转录组学。这些“组学”方法的整合使我能够 以获得对ETV 6的癌症维持功能的深入生物化学理解（目的1）。而且我 表明ETV 6的不育α基序（SAM）结构域的表达，该结构域负责其自身的表达。 寡聚化，对内源性ETV 6具有显性负效应，并抑制体内肉瘤发生。 因此，我建议优化这种SAM肽，以提高其效力，并进一步工程化其用于外源性 交付（目标2）。成功产生一种有效的、具有肿瘤穿透性的ETV 6阻滞剂将有利于治疗 发展最后，由于转录因子，如ETV 6，已被证明是药物治疗的具有挑战性的靶点， 在开发过程中，我分析了可以结合ETV 6以揭示可药用口袋的内源性代谢物， 鉴定了它与磷脂酸的关系。我将进一步探讨磷脂酸的调节作用 结合尤文肉瘤中的ETV 6（Aim 3）。这些研究的结果将指导小分子的开发， 更重要的是，它还将反映一种新的机制，通过直接代谢控制基因表达， 转录因子的变构调节。 在辅导K99阶段，我将与我的导师克里斯托弗Vakoc博士和共同导师博士密切合作。 Carolyn Fein Levy和合作者Stegmaier博士，Phassis博士，Shi博士和Furukawa博士，公认的儿科专家， 肿瘤学、肽治疗、筛选方法学和结构生物学。我还建立了 CSHL的一个特别咨询委员会，由Joshua-Tor博士和Beyaz博士组成，他们将监测和 支持我向独立过渡此外，CSHL将为我提供一个出色的科学环境 作为世界知名会议和课程的会议中心。我的目标 是获得一个教师职位，以制定一个有影响力的研究计划，其中K99/R 00资助机制 将成为我走向独立的重要一步