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: 8402605
• 负责人: Jennifer M Peterson
• 金额: $ 5.77万
• 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/8402605
• 关键词: Ablation; Adverse effects; Affect; Attenuated; Becker Muscular Dystrophy; Biological Assay; Cachexia; Cardiac; Cardiac Myocytes; Cardiomyopathies; Cause of Death; Cessation of life; ChIP-seq; Childhood; Data; Death Rate; Development; Disease; Disease Progression; Duchenne muscular dystrophy; Dystrophin; EMSA; Ensure; Female; Fibrosis; Gene Expression; Gene Targeting; Genes; Goals; Heart; Heart Diseases; Inflammation; Inflammatory; Malignant Neoplasms; Modeling; Mus; Muscular Dystrophies; Myocardium; NF-kappa B; Natural regeneration; Nuclear; Nuclear Protein; Pathology; Pathway interactions; Patients; Phosphotransferases; Quality of life; Regulation; Reporter; Reporting; Reverse Transcriptase Polymerase Chain Reaction; Rhabdomyosarcoma; Role; Signal Pathway; Signal Transduction; Skeletal Muscle; Steroids; Technology; Testing; Time; Tissues; Western Blotting; Work; base; cell type; effective therapy; functional group; gene therapy; genome-wide; improved; insight; mdx mouse; p65; public health relevance; skeletal; tool; transcription factor; wasting

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.

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。