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）：该提案的目标是测试从自然系统推断出的蛋白质相互作用原理是否可以用于蛋白质工程，以设计有用的基于促红细胞生成素（EPO）的分子。由于其刺激红细胞生成和抗炎途径，EPO 是治疗贫血和自身免疫性疾病的一种有吸引力的选择。然而，其临床应用受到其脱靶影响的抑制，包括 血栓形成和刺激肿瘤生长。为了满足对不同细胞类型具有更大特异性的基于 EPO 的药物的需求，可以找到一种合成生物学方法来研究细胞如何 使用多组分受体复合物特异性靶向细胞因子。例如，白细胞介素2受体由三个亚基组成，其中一个亚基充当另外两个亚基的高亲和力靶向结构域，以创建整体高亲和力受体[50]。我假设这种精确蛋白质靶向的天然机制可以转化为基于 EPO 的合成疗法的工程设计，改善对红系祖细胞或巨噬细胞的靶向，分别用于治疗贫血或自身免疫性疾病。我将构建嵌合蛋白，其中包含与其受体亲和力降低的 EPO 突变形式、与靶细胞牢固结合的靶向元件以及允许同时结合的接头。我假设，通过将结合动力学的定量信息与蛋白质配体和细胞表面受体的结构信息相结合，所得蛋白质（“嵌合激活剂”）应该仅在具有这两种元件受体的细胞中激活信号转导。靶向元件应介导初始结合，并且突变的 EPO 应随后结合，因为其在细胞表面的局部浓度较高，尽管其结合亲和力降低。这项工作将为设计基于蛋白质的靶向药物的定量方面、EPO 的多种生物学作用及其作为多效性治疗的价值提供有价值的信息，并且还可能为设计细胞类型特异性疗法提供新方法。总体而言，这些发现将对改善贫血或自身免疫性疾病患者的生活质量和疾病预后产生影响。