Thiol isomerases and ERO1α in sickle cell vaso-occlusion

镰状细胞血管闭塞中的硫醇异构酶和 ERO1α

• 批准号: 9977272
• 负责人: Jaehyung Cho
• 金额: $ 18.05万
• 依托单位: UNIVERSITY OF ILLINOIS AT CHICAGO
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-07-15 至 2020-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9977272
• 关键词: Acute; Adhesions; Adhesives; Affect; American; Animals; Attenuated; Binding; Binding Proteins; Biochemical; Biological; Biological Markers; Blocking Antibodies; Blood Cells; Blood Platelets; Blood specimen; Cause of Death; Cell Adhesion; Cell Aggregation; Cell Communication; Cell surface; Cells; Clinical Research; Complex; Complication; Data; ERp57; Endoplasmic Reticulum; Endothelial Cells; Estrogen receptor positive; Event; Extracellular Protein; Functional disorder; Goals; Hypoxia; Imaging Techniques; Inflammatory; Integrins; Isomerase; Ligand Binding; Ligands; Light; Macrophage-1 Antigen; Mediating; Microscopic; Molecular; Mus; Neutrophil Infiltration; Oxidases; Oxidation-Reduction; Oxides; Pain; Pathologic; Patients; Peptides; Peripheral; Plasma; Play; Protein Disulfide Isomerase; Proteins; Pulmonary vessels; Recurrence; Regulation; Role; Series; Sickle Cell; Sickle Cell Anemia; Signal Transduction; Structure; Sulfhydryl Compounds; Supporting Cell; Surface; Testing; Vascular Endothelium; Work; acute chest syndrome; base; design; disulfide bond; effective therapy; extracellular; improved; in vivo imaging; insight; mouse model; neutrophil; novel; novel therapeutics; preclinical study; real-time images; receptor; vascular inflammation

Project Summary Preclinical and clinical studies have provided compelling evidence that adherent neutrophils on activated vascular endothelium contribute to cell-cell aggregation and vaso-occlusion (VOC) in sickle cell disease (SCD). VOC positively correlates with severe pain crisis and acute chest syndrome, the common complication and cause of death in SCD patients. Currently, the effective treatment options for VOC remain limited. Thus, a better understanding of the regulatory mechanisms of neutrophil adhesive function will help develop novel therapies for treating VOC in SCD. We previously discovered that neutrophil surface-bound protein disulfide isomerase (PDI) is crucial for the ligand-binding activity of αMβ2 integrin and neutrophil-endothelial cell interactions during vascular inflammation. These findings have raised several fundamental questions regarding how the activity of cell surface-bound PDI is controlled and whether other thiol isomerases also regulate the function of neutrophil surface receptors. Our preliminary data demonstrate that endoplasmic reticulum protein 57 (ERp57, a thiol isomerase with structural similarity with PDI) and ER oxidoreductin 1α (ERO1α, a key oxidase of thiol isomerases in the ER) are detected on the surface of activated neutrophils and play overlapping and distinct roles in regulating neutrophil adhesive function under inflammatory conditions. Using biochemical, cellular, and animal studies with novel mouse models, blocking antibodies and cell-impermeable peptides, we will test the hypothesis that extracellular ERp57 and PDI in concert with ERO1α target different surface receptors and regulate the ligand-binding function, contributing to intravascular cell-cell interactions and VOC in SCD. In Aim 1, using a series of biochemical and cell biological studies, we will determine the molecular basis of neutrophil thiol isomerases and ERO1α in regulating neutrophil adhesive function. In Aim 2, using in vivo imaging techniques, we will test whether inhibition of extracellular ERp57, PDI and ERO1α attenuates intravascular cell-cell aggregation and VOC in peripheral and pulmonary vessels of SCD mice. In Aim 3, using blood samples from SCD patients under different VOC conditions, we will test whether extracellular thiol isomerases and ERO1α contribute to cell-cell aggregation and correlate with acute VOC conditions in SCD patients. Our studies will provide insights into novel molecular mechanisms mediating intravascular cell-cell interactions, which can be used to design safe and effective strategies to treat VOC-mediated conditions in SCD.

项目摘要 临床前和临床研究已经提供了令人信服的证据，表明粘附的中性粒细胞活化 血管内皮有助于镰状细胞病（SCD）中的细胞-细胞聚集和血管闭塞（VOC）。 VOC与疼痛危象、急性胸部综合征、常见并发症及病因呈正相关 SCD患者的死亡率。目前，VOC的有效治疗方案仍然有限。一个更好的 了解中性粒细胞粘附功能的调节机制将有助于开发新的治疗方法 用于处理SCD中的VOC。我们先前发现中性粒细胞表面结合蛋白二硫键异构酶 (PDI)对于αMβ2整联蛋白的配体结合活性和嗜中性粒细胞-内皮细胞相互作用至关重要， 血管炎症这些发现提出了几个基本问题，即 细胞表面结合的PDI是受控的，以及其他巯基异构酶是否也调节中性粒细胞的功能。 表面受体我们的初步数据表明，内质网蛋白57（ERp 57，一个巯基 与PDI结构相似异构酶）和ER氧化还原蛋白1α（ERO 1 α，巯基异构酶的关键氧化酶 在ER中）在活化的中性粒细胞表面上检测到，并在 在炎症条件下调节中性粒细胞粘附功能。利用生物化学，细胞，和动物 通过使用新型小鼠模型、阻断抗体和细胞不可渗透肽的研究，我们将检验这一假设 细胞外ERp 57和PDI与ERO 1 α协同作用，靶向不同的表面受体，并调节细胞内ERp 57和PDI的表达。 配体结合功能，有助于血管内细胞-细胞相互作用和SCD中的VOC。在目标1中，使用 通过一系列的生物化学和细胞生物学研究，我们将确定中性粒细胞巯基的分子基础 异构酶和ERO 1 α调节中性粒细胞粘附功能。在目标2中，使用体内成像技术， 我们将测试细胞外ERp 57、PDI和ERO 1 α的抑制是否会减弱血管内细胞-细胞间的相互作用， 聚集和VOC的SCD小鼠外周和肺血管。在目标3中，使用来自 SCD患者在不同VOC条件下，我们将检测细胞外巯基异构酶和ERO 1 α 有助于细胞-细胞聚集并与SCD患者的急性VOC状况相关。 我们的研究将提供新的分子机制介导的血管内细胞-细胞 相互作用，可用于设计安全有效的策略来治疗SCD中VOC介导的病症。