Relationship of candidate circulating proteoforms with aging phenotypes.

候选循环蛋白型与衰老表型的关系。

• 批准号: 9248089
• 负责人: RICHARD D SEMBA
• 金额: $ 21.06万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-06-01 至 2017-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9248089
• 关键词: 21 year old; Activities of Daily Living; Address; Adult; Affect; Age; Aging; Animal Model; Animals; Attention; Baltimore; Binding; Biological; Biological Assay; Blood; C-terminal; Carrier Proteins; Cleaved cell; Cognition; Conflict (Psychology); Development; Eotaxin; Epitopes; Follistatin; Follistatin-Like Protein 3; GDF11 gene; GDF8 gene; Genes; Health; Heart Hypertrophy; Human; Immunoassay; Insulin Resistance; Liquid Chromatography; Longitudinal Studies; Lower Extremity; Measures; Memory; Molecular; Monitor; Muscle; Oxytocin; Peptides; Phenotype; Physical Performance; Plasma; Post-Translational Protein Processing; Protein Isoforms; Proteins; Public Health; Reaction; Reagent; Rejuvenation; Reporting; Research; Research Design; Role; Skeletal Muscle; Stable Isotope Labeling; Technology; Testing; Validation; Walking; age related; aged; cohort; disability; growth-differentiation factor 8; inhibitor/antagonist; innovation; muscle form; muscle strength; neurophysins; novel; polypeptide; sarcopenia; secondary outcome; skeletal muscle growth; tandem mass spectrometry; therapeutic target; walking speed

 DESCRIPTION (provided by applicant): Recent animal studies have identified several polypeptides or proteins that can reverse or accelerate aging phenotypes. These candidates include growth/differentiation factor 11 (GDF11), growth/differentiation factor 8 (GDF8), oxytocin, eotaxin, and inhibitors of GDF11 and GDF8: GDF11 and GDF8 propeptides, follistatin, follistatin-related protein 3 (FSTR3), WFIKKN1, and WFIKKN2. These candidate polypeptides or proteins have been difficult to study in the blood using conventional immunoassays, since some of the peptides or proteins exist in multiple isoforms, undergo cleavage or terminal degradation, or have high sequence identity with each other. Whether these proteoforms (defined as the different molecular forms in which the protein product of a single gene can be found, such as isoforms, cleavage, and degradation products) have a similar relationship to aging phenotypes in humans is not known. We will use a novel multiplexed selected reaction monitoring (SRM) assay and liquid chromatography-tandem mass spectrometry (LC-MS/MS) to measure fifteen plasma proteoforms representing eight important proteins in rejuvenation research. We will test the hypothesis that plasma concentrations of these candidate proteoforms change with aging, and, beyond chronological age, predict the development of specific aging phenotypes in adults. The specific aims are: (1) to finalize development of the SRM assay using LC-MS/MS for absolute quantification of plasma proteoforms: GDF11 (propeptide, mature protein), GDF8 (propeptide, mature protein), follistatin (2 isoforms, 1 cleaved form), FSTR3, WFIKKN1, WFIKKN2, eotaxin, oxytocin (nonapeptide, 2 carboxyl-extended forms) and its carrier protein, neurophysin-1, (2) to characterize the relationship of circulating candidate proteoforms with aging phenotypes in the Baltimore Longitudinal Study of Aging and in a replication cohort, the InCHIANTI Study. Aging phenotypes include prevalent and incident sarcopenia, lower extremity physical performance, disability, cognition, memory, cardiac hypertrophy, and insulin resistance. By the end of the project, we should be able to verify or refute the role of these candidate proteoforms in phenotypes of human aging.

 描述（由申请人提供）：最近的动物研究已经鉴定了几种可以逆转或加速衰老表型的多肽或蛋白质。这些候选物包括生长/分化因子11（GDF 11）、生长/分化因子8（GDF 8）、催产素、嗜酸性粒细胞趋化因子以及GDF 11和GDF 8的抑制剂：GDF 11和GDF 8前肽、卵泡抑素、卵泡抑素相关蛋白3（FSTR 3）、WFIKKN 1和WFIKKN 2。这些候选多肽或蛋白质难以使用常规免疫测定法在血液中研究，因为一些肽或蛋白质以多种同种型存在，经历切割或末端降解，或彼此具有高序列同一性。这些蛋白质型（定义为不同的分子形式，其中可以发现单个基因的蛋白质产物，如同种型，切割和降解产物）是否与人类衰老表型有相似的关系尚不清楚。我们将使用一种新的多路选择反应监测（SRM）分析和液相色谱-串联质谱（LC-MS/MS）来测量代表复壮研究中8种重要蛋白质的15种血浆蛋白质型。我们将检验这一假设，即这些候选蛋白质型的血浆浓度随年龄的变化，并在实际年龄之外，预测成年人特定衰老表型的发展。具体目标是：（1）使用LC-MS/MS完成SRM测定法的开发，用于血浆蛋白质型的绝对定量：GDF11（前肽，成熟蛋白），GDF 8（前肽，成熟蛋白），卵泡抑素（2种亚型，1种裂解型）、FSTR 3、WFIKKN 1、WFIKKN 2、嗜酸性粒细胞趋化因子、催产素（九肽，2种羧基延伸形式）及其载体蛋白，神经垂体素-1，（2）在老化的巴尔的摩纵向研究和重复队列中表征循环候选蛋白质型与老化表型的关系，Inchianti研究衰老表型包括普遍和偶发的肌肉减少症、下肢体能、残疾、认知、记忆、心脏肥大和胰岛素抵抗。到项目结束时，我们应该能够验证或反驳这些候选蛋白质在人类衰老表型中的作用。