Targeting ligand-independent CSF3R dimerization in chronic neutrophilic leukemia

慢性中性粒细胞白血病中配体独立的 CSF3R 二聚化

• 批准号: 10011556
• 负责人: Michael Hollander
• 金额: $ 3.77万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-09-30 至 2021-09-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10011556
• 关键词: Affinity; Antibodies; Antineoplastic Agents; Binding; Biochemical; Biochemistry; Biological; Biology; CSF3 gene; Carbohydrates; Cell Survival; Cells; Chronic Neutrophilic Leukemia; Collaborations; Colony-Stimulating Factor Receptors; Complex; Crystallization; Crystallography; Dimerization; Directed Molecular Evolution; Disease; Dissociation; Electron Transport; Engineering; Epitopes; Family; Glycobiology; Goals; Granulocyte Colony-Stimulating Factor Receptors; Growth; In Vitro; Interleukin 6 Receptor; Isoleucine; Label; Learning; Length; Libraries; Ligands; Link; Malignant Neoplasms; Mass Spectrum Analysis; Metabolic; Modeling; Molecular; Mutate; Mutation; Outcome; Polysaccharides; Post-Translational Protein Processing; Protein Engineering; Proteins; Proteomics; Publishing; Rare Diseases; Research; Research Personnel; Role; Sampling; Serine; Signal Transduction; Site; Specificity; Structure; Testing; Therapeutic; Threonine; Training; Universities; Variant; Work; Yeasts; antibody engineering; base; cancer cell; cancer type; career; design; dimer; driver mutation; drug development; drug discovery; extracellular; glycosylation; in vitro Assay; in vivo; inhibitor/antagonist; mouse model; mutant; neoplastic cell; neutrophil; novel; novel therapeutics; receptor; skill acquisition; skills; small molecule; sugar; targeted treatment; therapeutic target; tool; translational cancer research

The modification of proteins with carbohydrates, called glycosylation, is commonly dysregulated in cancer. One example is chronic neutrophilic leukemia (CNL), a rare disease characterized by the uncontrolled growth of neutrophils. Over 80% of published CNL cases result from a mutation in the colony-stimulating factor 3 receptor (CSF3R). This mutation results in decreased O-linked glycosylation as well as increased ligand-independent dimerization. However, the relationship between these two findings is not known and there are no therapies that target the mutated epitope. The central hypothesis of this project is the loss of glycosylation decreases steric hindrance for CSF3R dimerization and reveals a cancer-specific epitope for targeted therapy. In Aim 1, mass spectrometry will identify the glycans that are present on the wild-type protein but missing in the mutated variant. Additionally, crystal structures of mutant CSF3R will reveal how the loss of glycosylation remodels the receptor interface and promotes ligand-independent dimerization. In Aim 2, screens of yeast-displayed proteins will identify candidates which bind to mutated, but not wild-type CSF3R. Biologics specific to the mutated protein will be validated for labeling cells and blocking dimerization in vitro, and selectively eliminating cancer cells in a mouse model of CNL. Combined, this work will demonstrate how understanding the structural effects of glycosylation facilitates drug discovery. This work is co-sponsored at Stanford University by Drs. Jennifer Cochran and Carolyn Bertozzi, leaders in protein engineering and glycobiology, respectively. This project is also supported through a collaboration with Dr. Julia Maxson, who first discovered the CSF3R mutation. This doctoral work will provide essential training for a research career, bridging the gap between molecular mechanisms of cancer and drug development.

蛋白质与碳水化合物的修饰，称为糖基化，通常在癌症中失调。一 例如慢性嗜中性粒细胞白血病（CNL），这是一种罕见的疾病，其特征是不受控制的生长， 中性粒细胞超过80%的已发表的CNL病例是由集落刺激因子3受体突变引起的 （CSF3R）。该突变导致O-连接糖基化降低以及配体非依赖性增加。 二聚化。然而，这两个发现之间的关系尚不清楚，也没有治疗方法， 靶向突变的表位。该项目的中心假设是糖基化的丧失降低了空间位阻。 CSF 3R二聚化的阻碍，并揭示了用于靶向治疗的癌症特异性表位。在目标1中，质量 光谱法将鉴定存在于野生型蛋白质上但在突变变体中缺失的聚糖。 此外，突变CSF 3R的晶体结构将揭示糖基化的丧失如何重塑受体 界面并促进配体非依赖性二聚化。在目标2中，酵母展示蛋白的筛选将 鉴定与突变的但不与野生型CSF 3R结合的候选物。针对突变蛋白质的生物制剂 在体外标记细胞和阻断二聚化，并选择性地消除癌细胞， CNL小鼠模型。结合起来，这项工作将展示如何理解的结构影响， 糖基化促进药物发现。这项工作是共同赞助的斯坦福大学博士詹妮弗 Cochran和Carolyn Bertozzi分别是蛋白质工程和糖生物学的领导者。该项目也是 通过与Julia Maxson博士的合作，他首先发现了CSF 3R突变。这个博士 这项工作将为研究生涯提供必要的培训，弥合分子机制之间的差距， 癌症和药物开发。