Role of Nucleoside-Diphosphate Kinase Signaling in Atrial Fibrillation

核苷二磷酸激酶信号传导在心房颤动中的作用

• 批准号: 10594130
• 负责人: Xander H.T. Wehrens
• 金额: $ 55.03万
• 依托单位: BAYLOR COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-02-06 至 2027-01-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10594130
• 关键词: Adaptor Signaling Protein; Animals; Ankyrin Repeat; Arrhythmia; Atrial Fibrillation; Binding; Biopsy; Ca(2+)-Calmodulin Dependent Protein Kinase; Calcium Signaling; Canis familiaris; Cell model; Chronic; Complex; Cyclic AMP; Cyclic AMP-Dependent Protein Kinases; Data; Dependovirus; Development; Event; Fluorescence Resonance Energy Transfer; G-Protein Signaling Pathway; G-Protein-Coupled Receptors; GTP-Binding Proteins; Genetic; Goals; Guanosine Triphosphate; Heart; Heart Atrium; Human; Knock-in Mouse; Maintenance; Mediating; Molecular; Morbidity - disease rate; Mus; Muscle Cells; Nucleoside diphosphate kinase B; Nucleoside-Diphosphate Kinase; Pathogenesis; Pathogenicity; Pathway interactions; Patients; Phosphoric Monoester Hydrolases; Phosphorylation; Phosphotransferases; Play; Predisposition; Prevention; Production; Protein Isoforms; Proteomics; Publishing; Role; RyR2; Ryanodine Receptor Calcium Release Channel; Sarcoplasmic Reticulum; Second Messenger Systems; Signal Transduction; Tertiary Protein Structure; Testing; Therapeutic; Validation; Work; canine model; designer receptors exclusively activated by designer drugs; gene therapy; improved; knock-down; mortality; mouse model; new therapeutic target; novel; novel therapeutic intervention; overexpression; preclinical study; prevent; receptor; regional difference; sensor

PROJECT SUMMARY / ABSTRACT Atrial fibrillation (AF), the most common arrhythmia, is associated with high morbidity and mortality, but remains difficult to treat due to an incomplete understanding of underlying mechanisms. Emerging evidence suggests that nucleoside diphosphate kinases (NDPKs) play an important role in the heart by being able to elevate cAMP levels in a G-protein receptor-independent manner. Our pilot data suggest that increased levels of NDPK-B and NDPK-C isoforms in patients with persistent (chronic) AF are associated with elevated cAMP levels, abnormal sarcoplasmic reticulum Ca2+ releases, ectopic (triggered) activity and inducible AF. The long- term goal of this project is to dissect the molecular and cellular basis of NDPK-dependent arrhythmia mechanisms in AF. The central hypothesis is that enhanced NDPK-B and -C levels promote AF by enhancing cAMP levels in the RyR2 microdomain, resulting in aberrant intracellular Ca2+ signaling that creates a substrate for AF initiation, maintenance, and progression. Three specific aims will be pursued: 1) Determine whether increased NDPK expression is both necessary and sufficient to promote spontaneous AF, 2) Assess whether elevated NDPK levels cause cAMP-dependent SR Ca2+ leak via RyR2, and 3) Determine the role of Ankrd1 within the RyR2-NDPK signaling complex. These studies will be performed in atrial myocytes from patients with AF, a dog model of AF, and various atrial-selective genetic mouse models of AF. Novel targeted cAMP FRET sensors and atrial-selective adeno-associated virus (AAV9)-mediated gene therapy will be utilized to resolve the pro-arrhythmic roles of NDPK-B/C and cAMP within specific cellular microdomains. These studies are expected to establish whether and how NDPK isoforms contributes to AF development and serve as a platform for the validation of NDPKs as novel druggable target for the prevention or treatment of AF.

项目概要/摘要 心房颤动 (AF) 是最常见的心律失常，与高发病率和死亡率相关，但 由于对潜在机制的不完全了解，治疗仍然很困难。新出现的证据 表明核苷二磷酸激酶（NDPK）能够在心脏中发挥重要作用 以不依赖于 G 蛋白受体的方式提高 cAMP 水平。我们的试点数据表明，水平增加 持续性（慢性）房颤患者的 NDPK-B 和 NDPK-C 亚型与 cAMP 升高相关 水平、异常肌浆网 Ca2+ 释放、异位（触发）活动和诱导性 AF。长- 该项目的长期目标是剖析 NDPK 依赖性心律失常的分子和细胞基础 AF 中的机制。核心假设是，NDPK-B 和 -C 水平的提高通过增强 NDPK-B 和 -C 水平来促进 AF RyR2 微域中的 cAMP 水平，导致异常的细胞内 Ca2+ 信号传导，从而产生底物 AF 的发生、维持和进展。将追求三个具体目标： 1) 确定是否 NDPK 表达增加对于促进自发性 AF 是必要且充分的，2) 评估是否 NDPK 水平升高导致 cAMP 依赖性 SR Ca2+ 通过 RyR2 泄漏，以及 3) 确定 Ankrd1 的作用 RyR2-NDPK 信号复合物内。这些研究将在患者的心房肌细胞中进行 AF、狗的 AF 模型以及各种心房选择性 AF 遗传小鼠模型。新型靶向 cAMP FRET 传感器和心房选择性腺相关病毒 (AAV9) 介导的基因治疗将用于 解决特定细胞微域内 NDPK-B/C 和 cAMP 的促心律失常作用。这些研究 预计将确定 NDPK 亚型是否以及如何促进 AF 发展并作为 验证 NDPK 作为预防或治疗 AF 的新型药物靶点的平台。