Alternative protein isoforms in ventricular remodeling

心室重构中的替代蛋白质亚型

• 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
• 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

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.

项目总结 许多心脏基因经历选择性剪接，以产生心脏与骨骼、成人与胎儿的异构体。 选择性剪接模式的差异调控是改变的遗传程序的中心组成部分 在缺血损伤后心脏重塑过程中。另一种剪接已被证明可以因果调节 心脏重构的严重性，因此代表了阻止疾病的潜在治疗靶点 进步。然而，尽管现在已经有大量可供选择的异构体转录本 在心脏基因组中发现的，关于蛋白质编码潜力的最新知识，因此是分子 这些替代亚型的功能较差。大多数已发现的异构体仍未在 蛋白质水平--因此，不同的亚型对蛋白质丰度、基序特征和 蛋白质之间的相互作用几乎是未知的。这一知识鸿沟阻碍了对……的更充分理解 选择性剪接对健康和疾病患者心肌的影响。 蛋白质组学技术近年来取得了进步，现在已经做好了转变的准备 心血管科学，但异型蛋白的检测仍然是一个巨大的挑战。我们和 其他人最近表明，转录组学(rna-seq)和蛋白质组学(质谱学)数据可能 结合在一起，以更深入地探索心脏蛋白质组。在本项目中，我们将结合 RNA-seq(模拟转录异构体)和蛋白质组学(提供蛋白质水平)各自的优势 证据)，以检查心脏选择性剪接。具体地说，我们将应用这个RNA-seq引导- 蛋白质组学方法：(I)确定胎儿和成人心脏中可供选择的蛋白质亚型的图景；(Ii) 利用小鼠心肌梗死和心肌梗死模型确定重塑心脏的疾病特征亚型 异丙肾上腺素刺激；(Iii)通过鉴定异构体来检测蛋白质异构体的功能后果。 疾病中特定的蛋白质-蛋白质相互作用和异构体表达的调节因子；以及(Iv)阐明 异构体特异性过表达/敲除对心肌细胞功能表型的影响。我们期待着 拟议研究的完成将为理解替代剪接的作用开辟新的途径 在心脏病方面。