NF-kappaB signaling in dystrophic cardiomyopathy

营养不良性心肌病中的 NF-κB 信号传导

• 批准号: 8003481
• 负责人: Jennifer M Peterson
• 金额: $ 5.22万
• 依托单位: OHIO STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-01-01 至 2013-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8003481
• 关键词: Ablation; Affect; Becker Muscular Dystrophy; Biological Assay; Cardiac Myocytes; Cardiomyopathies; Cessation of life; Childhood; Development; Disease; Disease Progression; Duchenne muscular dystrophy; Dystrophin; EMSA; Ensure; Gene Expression; Genes; Goals; Heart; Heart Diseases; Inflammatory; Modeling; Mus; Muscular Dystrophies; Myocardium; NF-kappa B; Nuclear; Nuclear Protein; Nuclear Proteins; Pathway interactions; Patients; Phosphotransferases; Public Health; Quality of life; Reporter; Reverse Transcriptase Polymerase Chain Reaction; Role; Signal Pathway; Signal Transduction; Skeletal Muscle; Time; Western Blotting; Work; cell type; effective therapy; functional group; genome-wide; improved; insight; mdx mouse; public health relevance; skeletal; transcription factor

DESCRIPTION (provided by applicant): Duchenne muscular dystrophy (DMD) is a deadly childhood disease that affects both skeletal and cardiac muscles and for which there is no effective treatment or cure. The majority of patients with DMD and those with a milder form of the disease, Becker muscular dystrophy (BMD) develop progressive cardiomyopathy that eventually leads to death in 20 and 50% of patients, respectively. We propose a mechanism of dystrophic cardiomyopathy disease progression that is dependent on signaling through the transcription factor nuclear factor-kappaB (NF-?B). Our previous work has demonstrated the requirement for NF-?B signaling in dystrophic skeletal muscles, however the implications of NF-?B signaling in dystrophic cardiac muscle remain unknown. In the proposed studies, Aim 1 will evaluate the relevance of NF-?B in dystrophic cardiomyopathy. Dystrophin- deficient mice (mdx) will be used as a model to determine how early NF-?B becomes activated, the specific NF-?B signaling pathway involved (classical vs. alternative), and the identity of cell types expressing NF-?B in dystrophic hearts. EMSA analysis, NF-?B reporter mice, immunostaining, western blotting, kinase assays, and FACS analysis will be used to evaluate Aim 1. Aim 2 will determine the requirement for NF-?B signaling in disease development. Mdx mice deficient for NF-?B signaling in all cell types and specifically in cardiomyocytes will be evaluated for histological, functional, and gene expression differences compared to mdx mice without NF-?B ablation. Aim 3 will investigate the mechanism by which NF-?B promotes dystrophic cardiomyopathy. Real time RT-PCR will be performed on cardiomyopathy-related genes that are known targets of NF-?B. Genes will be placed in functional groups (eg: fibrotic, inflammatory, survival) to determine if NF-?B is mechanistically promoting cardiomyopathy by targeting a specific group of genes. Additionally, a genome- wide search for all NF-?B targets will be performed using ChIP-sequencing to ensure an unbiased approach for evaluating NF-?B signaling targets is undertaken. The completion of these aims will provide significant insight into the role of the NF-?B signaling pathway in cardiomyopathy, as well as the relevance for targeting this signaling modulator for the treatment of DMD and BMD. PUBLIC HEALTH RELEVANCE: The overall goal of this project is to understand how the protein nuclear factor-kappaB (NF-:?B) is contributing to heart disease in Duchenne and Becker muscular dystrophy. Understanding the role of NF-?B signaling in the heart is critical for determining if we can target this pathway for therapy to slow disease progression and improve quality of life for patients.

描述（由申请人提供）：Duchenne肌肉营养不良（DMD）是一种致命的儿童疾病，会影响骨骼和心脏肌肉，并且没有有效的治疗或治愈。大多数DMD患者和患有疾病较轻的患者，贝克尔肌肉营养不良（BMD）患有进行性心肌病，最终分别导致20％和50％的患者死亡。我们提出了一种营养不良心肌病疾病进展的机制，该机制取决于通过转录因子核因子-Kappab（NF-？b）信号传导。我们以前的工作表明了营养不良骨骼肌中NF- b信号传导的要求，但是NF-？B信号在营养不良心脏肌肉中的含义仍然未知。在拟议的研究中，AIM 1将评估NF-？B在营养不良的心肌病中的相关性。肌营养不良缺乏小鼠（MDX）将用作模型，以确定NF-的早期激活，特定的NF-？B信号传导途径（经典与替代方案）以及在营养不良心中表达NF-的细胞类型的身份。 EMSA分析，NF-？B报告小鼠，免疫染色，蛋白质印迹，激酶测定和FACS分析将用于评估目标1。AIM2将确定疾病发育中NF-？B信号传导的要求。与没有NF-？B消融的MDX小鼠相比，将评估所有细胞类型中缺乏NF-？B信号传导的MDX小鼠。 AIM 3将研究NF-？B促进营养不良心肌病的机制。实时RT-PCR将对与NF-？b的已知靶标的心肌病相关基因进行。基因将放置在官能团（例如：纤维化，炎症，生存）中，以确定NF-？B是否通过靶向特定的基因来促进心肌病。此外，将使用芯片序列进行基因组进行广泛的搜索，以确保采用无偏见的方法来评估NF-？B信号传导目标。这些目标的完成将为NF- b信号通路在心肌病中的作用以及针对该信号调节器以治疗DMD和BMD的相关性。 公共卫生相关性：该项目的总体目标是了解蛋白质核因子-Kappab（NF - ：？b）如何在Duchenne和Becker肌肉营养不良中促进心脏病。了解NF-？B信号在心脏中的作用对于确定我们是否可以针对这种治疗途径来减缓疾病进展并改善患者的生活质量。