Nonreceptor tyrosine kinases in Systemic Lupus Erythematosus

系统性红斑狼疮中的非受体酪氨酸激酶

基本信息

批准号：
10610475
负责人：
W Todd MILLER
金额：
$ 31.56万
依托单位：
STATE UNIVERSITY NEW YORK STONY BROOK
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-05-25 至 2026-04-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10610475
关键词：
Active Sites Apoptosis Apoptotic Autoimmune Diseases BCAR1 gene Binding Binding Sites Biological Assay Cell Surface Receptors Cell model Cells Collaborations Coupled Coupling Crystallization Disease Environmental Risk Factor Enzyme Kinetics Enzymes Eukaryotic Cell Excision Fluorescence Genetic Growth Human Inflammatory Insecta Laboratories Link Lupus Macrophage Malignant Neoplasms Mammalian Cell Mammary Neoplasms Measures Mutation Normal Cell PTK6 gene Pathology Pathway interactions Patients Phagocytes Phagocytosis Phosphatidylserines Phosphotransferases Physiological Physiology Protein Tyrosine Kinase Proteins Receptor Protein-Tyrosine Kinases Regulation Role Series Signal Pathway Signal Transduction Structure Substrate Specificity Systemic Lupus Erythematosus Testing Work cohort experimental study human disease induced pluripotent stem cell insight loss of function mutation mutant new therapeutic target novel novel therapeutic intervention three dimensional structure

项目摘要

Tyrosine kinases are important regulators of growth, differentiation, and apoptosis in eukaryotic cells. Inappropriate tyrosine kinase signaling is observed in many cancers and inflammatory diseases. This project focuses on the structure, activity, and regulation of the Ack1 and Brk nonreceptor tyrosine kinases. In collaboration with Dr. Frederic Geissmann and colleagues, we have discovered a series of loss-of- function mutations in Ack1 and Brk in a cohort of patients with severe Systemic Lupus Erythematosus (SLE). These are the first tyrosine kinase mutations found to be linked to this disease. Our preliminary work shows that the mutations drastically decrease (or completely block) kinase activity, downstream signaling, and phagocytosis of apoptotic cells. We propose to conduct functional studies of the wild-type and mutant forms of Ack1 and Brk in mammalian cells, including experiments in induced pluripotent stem cell (iPSC)- derived macrophages from the lupus patients and their healthy relatives. We will carry out global screens of substrate specificity to determine whether the mutations “rewire” cellular signaling networks. We will also carry out mechanistic and structural studies on the purified mutant kinases, and test how they are coupled to the cell surface receptor MerTK. The overall hypothesis is that Ack1 and Brk link recognition of phosphatidylserine on apoptotic cells to phagocytic engulfment. Regulation or tuning of this signaling pathway could provide a new strategy for therapeutic approaches in autoimmune diseases.

酪氨酸激酶是真核细胞生长、分化和凋亡的重要调节因子。在许多癌症和炎症性疾病中观察到不适当的酪氨酸激酶信号传导。该项目重点研究 Ack1 和 Brk 非受体酪氨酸激酶的结构、活性和调节。通过与 Frederic Geissmann 博士及其同事的合作，我们发现了一系列丢失- 一组严重系统性红斑狼疮患者中 Ack1 和 Brk 的功能突变（SLE）。这是我们的初步工作发现的第一个酪氨酸激酶突变。显示突变显着降低（或完全阻断）激酶活性、下游信号传导、我们建议对野生型和突变型细胞进行功能研究。哺乳动物细胞中 Ack1 和 Brk 的形式，包括诱导多能干细胞 (iPSC) 的实验 - 我们将对来自狼疮患者及其健康亲属的巨噬细胞进行全球筛查。我们还将利用底物特异性来确定突变是否“重新连接”细胞信号网络。对纯化的突变激酶进行机制和结构研究，并测试它们如何耦合总体假设是 Ack1 和 Brk 连接识别。凋亡细胞上的磷脂酰丝氨酸对吞噬细胞的吞噬进行调节或调节。途径可以为自身免疫性疾病的治疗方法提供新的策略。