Discovery and Characterization of sORFs encoding mitochondrial peptides with a Role in Heart Failure

编码在心力衰竭中发挥作用的线粒体肽的 sORF 的发现和表征

• 批准号: 9977248
• 负责人: Emma K Murray
• 金额: $ 2.54万
• 依托单位: TEMPLE UNIV OF THE COMMONWEALTH
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-08-01 至 2021-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9977248
• 关键词: Algorithms; Area; Base Pairing; Base Sequence; Biological Process; Calcium; Cardiac; Cardiac Myocytes; Cardiovascular Diseases; Cardiovascular system; Cause of Death; Cell Death; Cessation of life; Clustered Regularly Interspaced Short Palindromic Repeats; Code; Computer Analysis; Coronary artery; Cost of Illness; Custom; DNA Repair; Data; Data Set; Databases; Development; Diagnosis; Diagnostic; Disease; Disease Progression; Economic Burden; Ensure; Exclusion; Future; Generations; Genes; Genetic; Genome; Genomics; Health Care Costs; Heart Injuries; Heart failure; Hela Cells; Human; Human Genome; Individual; Lead; Left; Life Expectancy; Ligation; Membrane Potentials; Mentors; Messenger RNA; Metabolic; Metabolic dysfunction; Methodology; Methods; Mitochondria; Modeling; Mus; Myocardial Infarction; Myocardial dysfunction; N-terminal; Neonatal; Open Reading Frames; Paper; Pathway interactions; Peptides; Physiological; Physiology; Play; Probability; Protein Sequence Analysis; Proteins; Quality of life; Role; Sampling; Scanning; Science; Sequence Analysis; Signal Transduction; Spectrum Analysis; Statistical Data Interpretation; Surgical Models; Technology; Tertiary Protein Structure; Therapeutic; Time; Training; Transcript; Translating; Validation; Ventricular; base; biophysical properties; cohort; collaborative environment; constriction; cost; deep sequencing; differential expression; genome-wide; health economics; improved; in silico; in vivo; loss of function; mRNA Expression; mitochondrial dysfunction; novel; novel therapeutics; overexpression; prediction algorithm; pressure; prevent; programs; small hairpin RNA; statistics; success; transcriptome sequencing

Project Summary The advent of genomic sequencing revolutionized the field of science, revealing the human genome sequence to be ~3 billion base pairs. The result of this venture was the identification of ~20,000 protein-coding genes, which compromise only ~1% of the genome. While ~10% of the genome is considered ‘silent’, the remaining ~89% is transcribed, but thought to be untranslated. Previous annotation methods resulted in the exclusion of transcripts under 300bp (100aa) due to the high rate of gene misidentification. Recent papers have discovered small open reading frames (sORFs) that encode peptides under 100aa with distinct biological functions. We hypothesize that there are numerous peptides with unknown biological functions that are prominent in cardiac physiology and disease. To identify novel sORFs we combined in vivo and in silico approaches, including a statistical prediction algorithm sORfinder, to compile a database of putative sORFs. Given the prominent role of mitochondrial dysfunction in heart failure (HF) we included mitochondrial-targeting prediction for all sORFs by employing n-terminal protein sequence analysis using the computational programs MitoFates and MitoProt. In addition, we have incorporated mRNA deep sequencing left ventricular samples of mice subjected to transaortic constriction (pressure-overload HF) or permanent ligation of the left coronary artery (myocardial infarction) at multiple stages of disease progression to identify differentially expressed sORFs in HF. To prioritize our search, we are ranking various components for unbiased target selection and experimental confirmation. We envision this novel database as having great importance within and beyond the cardiovascular field for identifying novel genes with therapeutic potential.

项目摘要 基因组测序的出现彻底改变了科学领域，揭示了人类基因组序列 大约30亿个碱基对。这项研究的结果是鉴定了约20，000个蛋白质编码基因， 仅占基因组的1%虽然约10%的基因组被认为是“沉默的”，其余的基因组被认为是“沉默的”。 约89%是转录的，但被认为是未翻译的。以前的注释方法导致排除 由于基因错误鉴定率高，转录本长度在300 bp（100 aa）以下。最近的论文发现 小的开放阅读框（sORF），编码100个氨基酸以下具有不同生物学功能的肽。我们 假设有许多具有未知生物学功能的肽在心脏中是突出的， 生理学和疾病。为了鉴定新的sORF，我们结合了体内和计算机方法，包括 统计预测算法sORfinder，以编译推定的sORF的数据库。鉴于其在 心力衰竭（HF）中线粒体功能障碍的研究中，我们纳入了所有sORF的靶向预测 通过使用计算程序MitoFates和MitoProt进行N-末端蛋白质序列分析。 此外，我们还整合了mRNA深度测序小鼠左心室样本， 经主动脉缩窄（压力超负荷HF）或永久性结扎左冠状动脉（心肌 在疾病进展的多个阶段中，对心肌梗死）进行分析，以鉴定HF中差异表达的sORF。到 优先考虑我们的搜索，我们正在对各种组件进行无偏见的目标选择和实验 确认.我们设想这个新的数据库具有非常重要的内部和外部的 心血管领域用于鉴定具有治疗潜力的新基因。