The Comparability and Reproducibility of Telomere Length Measurements for Population-based Studies

基于人群的研究中端粒长度测量的可比性和可重复性

• 批准号: 10017222
• 负责人: Idan Shalev
• 金额: $ 31.61万
• 依托单位: PENNSYLVANIA STATE UNIVERSITY, THE
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-09-13 至 2022-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10017222
• 关键词: 16 year old; Adult; Affect; African American; Age; Age-Years; Aging; Archives; Biological Aging; Biological Assay; Birth; Birth length; Blood; Blood Cell Count; Cell Aging; Cells; Cheek structure; Child; Child Abuse and Neglect; Chromosomes; Clinical Research; Collection; Comparative Study; Consensus; DNA; Data; Development; Disease; Etiology; Event; Funding; Future; Genes; Goals; Guidelines; Health; Individual; Infant; Interdisciplinary Study; Intervention Studies; Intramural Research Program; Laboratories; Laboratory Procedures; Length; Leukocytes; Life; Life Expectancy; Maintenance; Manuals; Measurement; Measures; Methodology; Methods; Mononuclear; Morbidity - disease rate; Mothers; National Institute of Child Health and Human Development; National Institute of Environmental Health Sciences; Newborn Infant; Nucleoproteins; Outcome; Peripheral; Peripheral Blood Mononuclear Cell; Persons; Phenotype; Play; Polymerase Chain Reaction; Population Heterogeneity; Population Research; Population Study; Predictive Factor; Procedures; Process; Prospective cohort; Protocols documentation; Recommendation; Reproducibility; Research; Research Personnel; Research Training; Risk Factors; Robotics; Role; Saliva; Sampling; Science; Spottings; Standardization; Swab; Systems Biology; Techniques; Testing; Tissue Sample; Tissues; Umbilical Cord Blood; Variant; Whole Blood; Work; age related; aged; cohort; cost; design; epidemiology study; ethnic diversity; field study; improved; innovation; interest; mortality; recruit; sample collection; sex; telomere; tissue processing

PROJECT SUMMARY Telomere length (TL) has been established as a marker for cellular aging and as one of the hallmarks of aging. Studies have implicated age-related TL with a broad range of risk factors that predict disease morbidity and early mortality. Researchers interested in telomere science are nevertheless concerned about precision measures and reproducibility, which is a burning issue in the field and the sciences more broadly. A key question concerns the measurement of TL from different types of tissue cells for large population- based studies. Blood has commonly been used in TL studies. However, blood collection is relatively invasive, costly, and limiting in terms of where and by whom it can be collected. Scientific progress in the understanding of how TL affects health and aging will be greatly enhanced if researchers are not limited to the use of blood. How TL differ within- and between-individuals across common tissue cells and developmental stages using the T/S ratio, monochrome multiplex and absolute quantitative PCR (qPCR) methods is not known. Moreover, technical laboratory variations such as sample collection, storage conditions and pipetting techniques can also influence TL measurements. Results of this study may reveal an optimal tissue cell for large population-based research, clarify the precision of pipetting techniques and other technical laboratory procedures for studies on TL, and enable comparisons with TL results derived using different measurement methods. Aim 1 will determine, in 5 different cohorts from birth to age 80, the comparability of TL across commonly sampled tissues [whole blood/leukocytes, peripheral blood mononuclear cells (PBMCs), dried blood spots (DBS), cord blood, saliva, and buccal cells] using multiple qPCR assays. Aim 2 will examine technical laboratory variation on TL measurement by testing fresh vs. archived tissues and DNA, as well as various sample collection procedures and pipetting techniques (i.e., manual vs. automated robotic). Aim 3 will test association and estimated effect sizes of potential TL variation associated with different tissues and technical lab techniques involving multiple system-biology and phenotypic measures. This study is significant because its rigorous design will enable strong inferences regarding the impact of tissue comparability from birth to 80 years on TL, as well as technical laboratory variations and estimated effect sizes of potential TL variation using multiple qPCR methods. Combined findings from the telomere consortium will help generate consensus guidelines for TL measurement. This study is innovative because it will be the first to comprehensively assess six tissues from the same sample across developmental stages and lab techniques, and will lay the groundwork for future comparisons. The results and raw data generated by this proposal will be shared with the Telomere Research Network consortium to increase transparency, standardize measurement methods, and facilitate cross-method comparison studies.

项目概要 端粒长度（TL）已被确立为细胞衰老的标志，也是细胞衰老的标志之一。 老化。研究表明，与年龄相关的 TL 与预测疾病发病率的多种危险因素有关 和早期死亡。然而，对端粒科学感兴趣的研究人员担心精度 测量和再现性，这是该领域和更广泛的科学领域的一个紧迫问题。 一个关键问题涉及大量人群不同类型组织细胞的 TL 测量 - 基础研究。 TL 研究中通常使用血液。然而，血液采集是相对侵入性的， 成本高昂，并且收集地点和收集人员受到限制。认识上的科学进步 如果研究人员不局限于血液的使用，那么关于 TL 如何影响健康和衰老的研究将会大大加强。 TL 如何在常见组织细胞和发育阶段的个体内部和个体之间使用 T/S 比、单色多重和绝对定量 PCR (qPCR) 方法尚不清楚。而且， 技术实验室变化，例如样品采集、储存条件和移液技术也可能 影响 TL 测量。这项研究的结果可能揭示基于大量人群的最佳组织细胞 研究，阐明移液技术和其他技术实验室程序的精度，以进行研究 TL，并可以与使用不同测量方法得出的 TL 结果进行比较。 目标 1 将确定 5 个不同队列（从出生到 80 岁）的 TL 的可比性 组织样本[全血/白细胞、外周血单核细胞 (PBMC)、干血斑 (DBS)、脐带血、唾液和口腔细胞]使用多种 qPCR 检测。目标 2 将检查技术 通过测试新鲜与存档的组织和 DNA 以及各种 样品采集程序和移液技术（即手动与自动机器人）。目标 3 将进行测试 与不同组织和技术相关的潜在 TL 变异的关联和估计效应大小 涉及多种系统生物学和表型测量的实验室技术。 这项研究意义重大，因为其严格的设计将能够就以下因素的影响做出强有力的推论： 从出生到 80 岁的 TL 组织可比性，以及技术实验室变化和估计效果 使用多种 qPCR 方法确定潜在 TL 变异的大小。端粒联盟的综合发现 将有助于形成 TL 测量的共识指南。这项研究具有创新性，因为它将 第一个全面评估同一样本中不同发育阶段和实验室的六种组织 技术，并为将来的比较奠定基础。由此生成的结果和原始数据 该提案将与端粒研究网络联盟共享，以提高透明度、标准化 测量方法，并促进跨方法比较研究。