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.

项目摘要/摘要 尤文肉瘤是一种高度侵袭性的骨和软组织肿瘤，主要影响儿童和年轻人， 仍然缺乏有效的治疗方法。由于它的变异负担很低，因此在 尤文肉瘤细胞。在我的博士后工作中，我发现转录因子ETV6是 极少有尤因肉瘤偏向的依赖。值得注意的是，尽管ETV6表达无处不在，但它并没有 尤文肉瘤以外的癌细胞的必需品，基于我们和其他人的CRISPR筛查努力。 对小鼠的遗传学研究也表明，ETV6在大多数组织中都是必不可少的。因此，心烦意乱 ETV6在尤文肉瘤中的作用将具有广泛的治疗指标。在拟议的项目中，我的目标是进一步 研究ETV6如何维持癌细胞状态的详细机制，并利用这一新知识 开发对尤文肉瘤患者具有非凡效力和特异性的治疗方法。为此，我将 ETV6缺陷型尤文肉瘤细胞间充质分化表型的研究 吞吐量功能基因组学和单细胞转录组学。整合这些“经济学”方法可以让我 深入了解ETV6的癌症维持功能(目标1)。此外，我 研究表明，ETV6的不育α基序(SAM)结构域的表达与其自身的 寡聚，对内源性ETV6有显性-负性作用，抑制体内肉瘤形成。 因此，我建议对该SAM多肽进行优化，以提高其效力，并进一步将其用于外源 交付(目标2)。成功研制出一种有效的、可穿透肿瘤的ETV6阻滞剂将有利于治疗 发展。最后，由于转录因子，如ETV6，已被证明是具有挑战性的药物靶点 开发时，我分析了可以结合ETV6的内源性代谢物，以揭示可用药口袋，以及 确定了它与磷脂酸的联系。我将进一步探讨磷脂酸的调节作用。 尤文肉瘤与ETV6的结合(AIM 3)。这些研究的结果将指导小分子的开发， 更重要的是，这也将反映一种新的代谢控制基因表达的机制 转录因子的变构调节。 在指导K99阶段，我将与我的导师Christopher Vakoc博士和共同导师Dr。 Carolyn Fein Levy及其合作者Stegmaier、Kentsis、Shih和Furukawa博士是公认的儿科专家 肿瘤学、多肽疗法、筛查方法学和结构生物学。我还确立了 CSHL的一个特殊咨询委员会，由Joshua-Tor博士和Beyaz博士组成，他们将监督和 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 对于我的研究和培训，成为世界知名会议和课程的会议中心。我的目标 是获得一个教职员工的职位来开发一个有影响力的研究计划，其中K99/R00的资助机制 将成为我向独立过渡的重要一步。