A synthetic biology approach to targeting erythropoietin-based therapeutics

一种基于促红细胞生成素的靶向治疗的合成生物学方法

• 批准号: 8649234
• 负责人: Devin R. Burrill
• 金额: $ 4.99万
• 依托单位: HARVARD MEDICAL SCHOOL
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-02-01 至 2017-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8649234
• 关键词: Accounting; Acute Erythroblastic Leukemia; Address; Affect; Affinity; Anemia; Animals; Anti-Inflammatory Agents; Anti-inflammatory; Autoimmune Diseases; Behavior; Binding; Biological; Biological Assay; Blood Platelets; Cell Line; Cell Proliferation; Cell Surface Receptors; Cell surface; Cells; Chimeric Proteins; Clinical; Complex; Disease; Drug Kinetics; Elements; Engineering; Erythroid; Erythropoiesis; Erythropoietin; Erythropoietin Receptor; Gene Activation; Glycophorin A; Goals; Hematology; IACUC; In Vitro; Inflammatory Response; Interleukin 2 Receptor; Kinetics; Length; Ligands; Measurement; Mediating; Methods; Mus; Mutate; Pathway interactions; Patients; Pharmaceutical Preparations; Pharmacodynamics; Procedures; Property; Protein Binding; Protein Engineering; Proteins; Protocols documentation; Quality of life; Role; Signal Transduction; Space Perception; Specificity; Structure; Surface; System; Testing; Therapeutic; Thrombosis; Translating; Work; Writing; base; cell type; cytokine; design; flexibility; improved; in vitro activity; in vivo; macrophage; outcome forecast; progenitor; public health relevance; receptor; synthetic biology; tumor growth

DESCRIPTION (provided by applicant):The goal of this proposal is to test whether principles of protein interaction that have been inferred from natural systems can be used in protein engineering to design useful erythropoietin (EPO)-based molecules. Due to its stimulation of erythropoiesis and anti-inflammatory pathways, EPO is an attractive treatment option for anemia and autoimmune disease. However, its clinical use is inhibited by its off-target affects, including thrombosis and stimulation of tumor growth. To address the need for EPO-based drugs with greater specificity for distinct cell types, a synthetic biology approach can be found in how cells use multi-component receptor complexes to specifically target cytokines. For example, the interleukin 2 receptor is composed of three subunits, one of which acts as a high-affinity targeting domain for the other two to create an overall high-affinity receptor [50]. I hypothesize that this natural mechanism for precise protein targeting can be translated to the engineering of synthetic EPO-based therapeutics with improved targeting of erythroid progenitors or macrophages, for the treatment of anemia or autoimmune disease, respectively. I will build chimeric proteins composed of a mutated form of EPO with reduced affinity for its receptor, a targeting element that binds strongly to the target cell, and a linker that allows simultaneous binding. I hypothesize that by combining quantitative information about binding kinetics with structural information about protein ligands and cell surface receptors, the resulting proteins ("chimeric activators") should activate signal transduction only in cells with receptors for both elements. The targeting element should mediate initial binding, and the mutated EPO should subsequently bind due to its high local concentration at the cell surface, despite its decreased binding affinity. This work will provide valuable information about the quantitative aspects of designing targeted protein-based drugs, the diverse biological roles of EPO and its value as a pleiotropic therapeutic, and may also enable new methods for engineering cell type-specific therapies. Overall, the findings will have implications for improving the quality of life and diseae prognosis in patients suffering from anemia or autoimmune disease.

描述（由申请人提供）：该提案的目的是测试是否可以在蛋白质工程中使用自然系统推断的蛋白质相互作用原理来设计有用的促红细胞生成素（EPO）基于基于的分子。由于刺激了红细胞生成和抗炎途径，EPO是贫血和自身免疫性疾病的有吸引力的治疗选择。但是，它的临床用途受到其脱靶影响的抑制 血栓形成和肿瘤生长的刺激。为了满足对不同细胞类型具有更特异性的基于EPO的药物的需求，可以在细胞中找到一种合成生物学方法 使用多组分受体复合物特定靶向细胞因子。例如，白细胞介素2受体由三个亚基组成，其中一个是另外两个亚基的高亲和力靶向结构域，以创建一个总体高亲和力受体[50]。我假设这种用于精确蛋白质靶向的自然机制可以转化为基于合成的EPO疗法的工程，并改善了靶向红细胞祖细胞或巨噬细胞的靶向，以治疗贫血或自身免疫性疾病。我将构建由突变形式的EPO组成的嵌合蛋白，其对其受体的亲和力降低，其靶向元件与靶细胞强烈结合，以及允许同时结合的接头。我假设，通过将有关结合动力学的定量信息与有关蛋白质配体和细胞表面受体的结构信息相结合，所得蛋白（“嵌合激活剂”）应仅在具有两个元素的受体中激活细胞中的信号转导。靶向元件应介导初始结合，尽管其在细胞表面的局部浓度较高，但突变的EPO应随后结合，尽管其结合亲和力降低。这项工作将提供有关设计靶向蛋白质药物的定量方面，EPO的多种生物学作用及其作为多效性治疗的价值的有价值的信息，还可能启动用于工程细胞类型特异性疗法的新方法。总体而言，这些发现将对患有贫血或自身免疫性疾病的患者的生活质量和疾病预后有影响。