Defining and targeting the molecular vulnerabilities of the PAX3-FOXO1 protein in rhabdomyosarcoma

横纹肌肉瘤中 PAX3-FOXO1 蛋白的分子脆弱性的定义和靶向

• 批准号: 10221081
• 负责人: CHRISTOPHER M COUNTER
• 金额: $ 3.81万
• 依托单位: DUKE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-08-11 至 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10221081
• 关键词: Affect; Alveolar Rhabdomyosarcoma; Animal Model; Automobile Driving; Belief; Binding; Biological; Biological Assay; Biology; CRISPR screen; Cancer Biology; Cell Line; Cell model; Chemicals; Child; Chimeric Proteins; Chromatin; Clinical; Code; Collaborations; Data; Data Analyses; Dependence; Development; Disease; Elements; Environment; FOXO1A gene; Fusion Oncogene Proteins; Gene Expression; Gene Proteins; Genes; Genetic; Genetic Screening; Genetic Transcription; Genetically Engineered Mouse; Genomics; Glean; Goals; Human; In Vitro; Institution; Knowledge; Label; Lead; Leadership; Malignant Childhood Neoplasm; Mediating; Molecular Genetics; Molecular Target; Mutagenesis; Oncogenic; Oncoproteins; PAX3 gene; Pharmaceutical Chemistry; Pharmacology; Phenotype; Preparation; Protac; Proteins; Proteomics; Regulatory Element; Research; Research Personnel; Resolution; Resources; Rhabdomyosarcoma; Specialist; Speed; Structure; Structure-Activity Relationship; Techniques; Therapeutic; Therapeutic Intervention; Untranslated RNA; Validation; cellular engineering; clinical care; combinatorial; druggable target; epigenomics; genome-wide; improved; in vivo; innovation; insight; mouse model; multidisciplinary; neoplastic cell; novel; pre-clinical; public health relevance; small molecule; stem; structural biology; targeted agent; tool; transcription factor; transcriptomics; treatment strategy; tumor; tumorigenesis; tumorigenic

ABSTRACT – Overall (revised) Fusion-positive alveolar rhabdomyosarcoma (ARMS) remains one of the most fatal but least understood cancers of childhood. The driving oncoprotein in ARMS is the PAX3-FOXO1 fusion protein, a chimeric transcription factor that hijacks normal gene expression and chromatin state. Five-year survival for children with PAX3-FOXO1- positive ARMS is ~30% for all-comers, and <10% when metastatic. Despite the discovery of PAX3-FOXO1 in 1993, treatment strategies for affected children remain unchanged. This deficiency stems equally from a lack of understanding of the basic biology of the disease and an inability to directly target the fusion protein. No systematic or comprehensive approach has been undertaken to identify the proteins and regulatory elements required to support PAX3-FOXO1-mediated tumorigenesis. As a result, the field has been limited to a patchwork of data with no unified scientific strategy. To overcome this, this FusOnC2 Center has an innovative team and dynamic environment in which data interpretation is informed by complementary technological approaches and by biological and clinical knowledge. This comprehensive approach will transform understanding of PAX3-FOXO1-mediated oncogenesis and create opportunities for therapeutic intervention. The Center’s overarching goal is to advance the therapeutic tractability of the PAX3-FOXO1 fusion protein in ARMS by comprehensively identifying the druggable co-regulators, modulators, and intrinsic activities of PAX3-FOXO1. To accomplish this goal, the Center includes two complementary Projects, each led by expert RMS biologists paired with specialists in pioneering experimental approaches undertaking the most cutting-edge research in cancer biology, genomics, proteomics, structural biology, and medicinal chemistry. The Projects will be supported by RMS investigators within each project who will provide curated RMS cell lines, unique human primary RMS tumor cells, and murine models to enable rapid in vitro and in vivo validation and cross-prioritization of targets. An Administrative Core will integrate and coordinate the Center components, providing leadership and oversight, and promoting cross-pollination of ideas and resources. The Overall Specific Aims are to: (1) define and target the PAX3-FOXO1 interactome; and (2) perform chemical probe discovery for PAX3-FOXO1 to create additional tools for investigating the fundamental biology and tractability of PAX3-FOXO1 and fusion-positive RMS. Approaches used include proximity labeling, saturation mutagenesis, single and combinatorial CRISPR screens, high-throughput phenotypic assays, and mechanistically unbiased approaches to chemical probe discovery using novel high-throughput binding assays. We will use information gleaned to prioritize targets and agents for validation and to inform compound optimization and PROTAC preparation. This Center’s strengths and resources will synergize with the FusOnC2 Consortium, speeding development of knowledge generalizable to the biology of multiple fusion oncoproteins in childhood cancers, accelerating advances in clinical care.

摘要 - 总体（修订） 融合阳性肺泡横纹肌肉瘤（ARM）仍然是最致命但最不知情的童年癌症之一。手臂中驱动的癌蛋白是PAX3-福克斯1融合蛋白，这是一种嵌入正常基因表达和染色质状态的嵌合转录因子。 PAX3-FoxO1阳性臂的儿童的五年生存率约为30％，转移性时为<10％。尽管在1993年发现了PAX3-FoxO1，但受影响儿童的治疗策略仍然没有变化。这种缺乏阶段同样缺乏对疾病的基本生物学以及无法直接靶向融合蛋白的理解。没有采用系统或全面的方法来识别支持PAX3-福克斯介导的肿瘤发生所需的蛋白质和调节因素。结果，该领域仅限于没有统一的科学策略的数据拼凑而成。为了克服这一点，这个Fusonc2中心拥有一个创新的团队和动态环境，其中数据解释通过完整的技术方法以及生物学和临床知识来告知。这种全面的方法将改变对PAX3-FOXO1介导的肿瘤发生的理解，并为治疗干预创造机会。该中心的总体目标是通过全面识别PAX3-FoxO1的可药物共同调节剂，调节剂和内在活性来提高ARM中PAX3-FoxO1融合蛋白的治疗障碍性。为了实现这一目标，该中心包括两个完整的项目，每个项目都由专家RMS生物学家领导，并与专家一起采用了开创性的实验方法，从事癌症生物学，基因组学，蛋白质组学，结构生物学和医学化学方面最尖端的研究。这些项目将得到每个项目中的RMS研究人员的支持，他们将提供策划的RMS细胞系，独特的人类原发性RMS肿瘤细胞以及鼠模型，以实现靶标的快速体外和体内验证和交叉优先级。行政核心将整合和协调中心组件，提供领导和监督，并促进思想和资源的交叉授粉。总体具体目的是：（1）定义和靶向PAX3-FoxO1相互作用组； （2）对Pax3-FoxO1进行化学证明，以创建用于研究PAX3-FoxO1和融合阳性RMS的基本生物学和障碍的其他工具。所使用的方法包括接近性标记，饱和诱变，单一和组合CRISPR筛选，高通量表型测定以及使用新型的高通量结合测定法进行化学探针发现的机械无偏见方法。我们将使用收集的信息来确定目标和代理以验证并为复合优化和Protac准备提供信息。该中心的优势和资源将与Fusonc2联盟协同作用，从而加快了可推广到儿童癌中多种融合癌蛋白生物学的知识发展，从而加速了临床护理的进步。