Development of high-throughput cellular models for ASXL1-related diseases

项目摘要 我们广泛的生物学目标是开发一种基于细胞的模型，可用于研究分子发病机制 ASXL 1（Additional Sex Combs Like 1）突变引起的药物反应。额外的性喜欢（ASXL） 基因是一个基因家族的一部分，这些基因最初被鉴定为在细胞命运决定中起作用， 胚胎发育从头开始，截短突变ASXL 1导致儿科综合征，Bohring-Opitz 综合征（OMIM#605039）。ASXL 1突变也是急性髓细胞白血病的驱动突变。尽管 ASXL 1在多种疾病发病机制中的明确作用，其分子功能仍不清楚， 靶向药物已被批准用于患者。这一建议建立在我们的工作基础之上， 推定的表观遗传、RNA和蛋白质生物标志物一致地被ASXL 1中的致病性突变改变。我们 将使用基因组编辑方法将高度敏感的内源性标签引入我们的蛋白质中， 兴趣我们将专注于将这些标签引入到诱导多能干细胞系中，因为这些细胞系能够 在多种细胞类型中分化，这些细胞类型反映了人类疾病中受影响的关键器官系统。这些 蛋白质标签将使我们能够研究ASXL 1的分子和细胞功能，并为未来开发细胞模型 药物筛查该提案将满足罕见疾病的关键需求：开发基于细胞的模型系统， 确定可以快速识别ASXL 1突变病变细胞中观察到的致病分子变化的药物。

PROJECT ABSTRACT Our broad biological goal is to develop a cell-based model that can be used to study the molecular pathogenesis and drug responses due to mutations in ASXL1 (Additional Sex Combs Like 1). Additional Sex Like (ASXL) genes are part of a family of genes that were originally identified as having a role in cell-fate determination in the developing embryo. De novo, truncating mutations ASXL1 cause the pediatric syndrome, Bohring-Opitz Syndrome (OMIM#605039). Mutations in ASXL1 are also driver mutations in acute myeloid leukemia. Despite the clear role in multiple disease pathogenesis, the molecular function of ASXL1 remain unknown and no targeted drugs have been approved for use in patients. This proposal builds on our work which has identified putative epigenetic, RNA and protein biomarkers consistently altered by pathogenic mutations in ASXL1. We will use genome-editing approaches to introduce highly sensitive and endogenous tags to our proteins-of- interest. We will focus on introducing these tags into induced pluripotent stem cell lines as these are able to be differentiated in multiple cell-types that are reflective of key organ systems affected in human disease. These protein tags will allow us to study ASXL1-molecular and cellular function and to develop cell models for future drug screens. This proposal will meet a critical need in rare disease: to develop a cell-based model system to identify drugs that can rapidly identify pathogenic molecular changes observed in ASXL1-mutated diseased cells.