Alternative protein isoforms in ventricular remodeling

心室重构中的替代蛋白质亚型

基本信息

批准号：
10391342
负责人：
Maggie Lam
金额：
$ 38.91万
依托单位：
UNIVERSITY OF COLORADO DENVER
依托单位国家：
美国
项目类别：
财政年份：
2018
资助国家：
美国
起止时间：
2018-06-01 至 2023-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10391342
关键词：
Adult Affinity Chromatography Aging Alternative Splicing Amino Acid Sequence Biological Assay Biological Phenomena Cardiac Cardiovascular Diseases Cardiovascular system Cell Death Cell Death Process Code Coupled Data Detection Development Disease Disease Progression Event Exons Foundations Genes Genetic Genome Goals Guide RNA Health Heart Heart Diseases Heart Hypertrophy Human Hypertrophy In Vitro Isoproterenol Knowledge Level of Evidence Malignant Neoplasms Mass Spectrum Analysis Messenger RNA Metabolic Metabolism Modeling Molecular Muscle Cells Myocardial Infarction Myocardial Ischemia Myocardium Neonatal Oxygen Pathogenesis Pathology Pattern Perinatal Phenotype Positioning Attribute Protein Isoforms Proteins Proteome Proteomics RNA Splicing Rattus Regulation Research Resolution Role Sampling Science Severities Techniques Technology Transcript Translating Ventricular Ventricular Remodeling crosslink differential expression fetal heart function in silico ischemic injury knock-down molecular sequence database mouse model multiple omics novel strategies overexpression programs protein function protein protein interaction ribosome profiling screening skeletal therapeutic target tool transcriptome transcriptome sequencing transcriptomics virtual

项目摘要

PROJECT SUMMARY Many cardiac genes undergo alternative splicing to produce cardiac vs. skeletal, and adult vs. fetal isoforms. Differential regulation of alternative splicing patterns is a central component of the altered genetic program during cardiac remodeling after ischemic injuries. Alternative splicing has been shown to causally modulate the severity of cardiac remodeling, and thus represents a potential therapeutic target to halt disease progression. Nevertheless, although a large number of alternative isoform transcripts have now been discovered in the cardiac genome, current knowledge on the protein coding potential and hence molecular functions of these alternative isoform is poor. The majority of discovered isoforms were yet to be detected at the protein level -- therefore the effects of alternative isoforms on protein abundance, motif features, and protein-protein interactions are virtually unknown. This knowledge gap obstructs a fuller understanding on the impact of alternative splicing on the myocardium in health and disease. Proteomics technologies have advanced in recent years and are now well positioned to transform cardiovascular sciences, but the detection of isoform proteins remains a substantial challenge. We and others have recently shown that transcriptomics (RNA-seq) and proteomics (mass spectrometry) data may be combined to probe more deeply into the cardiac proteome. In this project, we will combine the respective strengths of RNA-seq (modeling transcript isoforms) and proteomics (providing protein-level evidence) to examine cardiac alternative splicing. Specifically, we will apply this RNA-seq-guided- proteomics approach to: (i) define the landscape of alternative protein isoforms in fetal and adult hearts; (ii) identify disease signature isoforms in remodeling hearts using mouse models of myocardial infarction and isoproterenol stimulation; (iii) examine the functional consequence of protein isoforms by identifying isoform- specific protein-protein interactions and modulators of isoform expression in disease; and (iv) elucidate the impact of isoform-specific overexpression/knockdown on myocyte functional phenotypes. We anticipate the completion of the proposed studies will open new avenues into understanding the role of alternative splicing in heart diseases.

项目摘要许多心脏基因经历替代剪接以产生心脏与骨骼，以及成人与胎儿同工型。替代剪接模式的差异调节是改变遗传程序的核心组成部分缺血性损伤后心脏重塑期间。替代剪接已显示为因果调节心脏重塑的严重程度，因此代表了停止疾病的潜在治疗靶点进展。然而，尽管现在已经有大量替代同工型转录本在心脏基因组中发现，有关蛋白质编码潜力的当前知识，因此是分子的这些替代同工型的功能很差。大多数发现的同工型尚未检测到蛋白质水平 - 因此，替代同工型对蛋白质丰度，基序特征和蛋白质 - 蛋白质相互作用实际上是未知的。这种知识差距阻碍了对替代剪接对健康和疾病心肌的影响。蛋白质组学技术近年来已经进步，现在已经有好处转变心血管科学，但同工蛋白的检测仍然是一个重大挑战。我们和其他人最近表明，转录组学（RNA-SEQ）和蛋白质组学（质谱法）数据可能将其组合成更深入的心脏蛋白质组。在这个项目中，我们将结合 RNA-seq（建模成绩单同工型）和蛋白质组学（提供蛋白质水平）的各自强度证据）检查心脏替代剪接。具体而言，我们将应用此RNA-Seq引导 - 蛋白质组学方法：（i）定义胎儿和成人心脏中替代蛋白质同工型的景观；（ii）使用心肌梗死的小鼠模型和异丙肾上腺素刺激；（iii）通过鉴定同工型来检查蛋白质同工型的功能后果疾病中同工型表达的特定蛋白质蛋白相互作用和调节剂；（iv）阐明同工型特异性过表达/敲低对心肌功能表型的影响。我们期待的拟议研究的完成将为理解替代剪接的作用开放新的途径在心脏病中。