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: 10221086
• 负责人: CHRISTOPHER M COUNTER
• 金额: $ 8.05万
• 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: 2020-08-12 至 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10221086
• 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（ARMS）仍然是儿童时期最致命但最不为人所知的癌症之一。 ARMS 中的驱动癌蛋白是 PAX3-FOXO1 融合蛋白，这是一种嵌合转录因子，可以劫持正常基因表达和染色质状态。所有 PAX3-FOXO1 阳性 ARMS 儿童的五年生存率约为 30%，转移时<10%。尽管 PAX3-FOXO1 于 1993 年被发现，但针对受影响儿童的治疗策略仍未改变。这种缺陷同样源于对该疾病的基本生物学缺乏了解以及无法直接靶向融合蛋白。目前尚未采取系统或全面的方法来鉴定支持 PAX3-FOXO1 介导的肿瘤发生所需的蛋白质和调控元件。结果，该领域仅限于拼凑的数据，没有统一的科学策略。为了克服这一问题，该 FusOnC2 中心拥有一支创新团队和动态环境，通过互补的技术方法以及生物学和临床知识来解释数据。这种综合方法将改变对 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鉴定 PAX3-FOXO1 的可药物协同调节剂、调节剂和内在活性，提高 ARMS 中 PAX3-FOXO1 融合蛋白的治疗易处理性。为了实现这一目标，该中心包括两个互补的项目，每个项目均由 RMS 生物学家专家领导，并与开拓实验方法的专家合作，开展癌症生物学、基因组学、蛋白质组学、结构生物学和药物化学领域最前沿的研究。这些项目将得到每个项目内 RMS 研究人员的支持，他们将提供精心策划的 RMS 细胞系、独特的人类原代 RMS 肿瘤细胞和小鼠模型，以实现快速的体外和体内验证以及目标的交叉优先级排序。行政核心将整合和协调中心各组成部分，提供领导和监督，并促进思想和资源的交叉授粉。总体具体目标是： (1) 定义并瞄准 PAX3-FOXO1 相互作用组； (2) 对 PAX3-FOXO1 进行化学探针发现，以创建其他工具来研究 PAX3-FOXO1 和融合阳性 RMS 的基础生物学和易处理性。使用的方法包括邻近标记、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高通量结合测定的机械无偏化学探针发现方法。我们将利用收集到的信息来确定目标和药物的优先顺序以进行验证，并为化合物优化和 PROTAC 制备提供信息。该中心的优势和资源将与 FusOnC2 联盟发挥协同作用，加快儿童癌症中多种融合癌蛋白生物学知识的发展，加速临床护理的进步。