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

项目总结 端粒长度(TL)已被确定为细胞衰老的标志和标志之一 衰老。研究表明，与年龄相关的TL与预测疾病发病率的一系列风险因素有关 和早期死亡率。然而，对端粒科学感兴趣的研究人员担心的是精确度 测量和可再现性，这是该领域和更广泛的科学界亟待解决的问题。 一个关键问题涉及到从不同类型的组织细胞测量大量人群的热释光。 基础研究。血液通常用于TL研究。然而，采血相对有侵入性， 成本高昂，而且在收集地点和收集者方面存在限制。认识中的科学进步 如果研究人员不局限于血液的使用，那么TL如何影响健康和衰老的研究将大大加强。 在常见的组织细胞和发育阶段中，TL在个体内和个体之间如何不同 T/S比率、单色多重和绝对定量聚合酶链式反应(QPCR)方法尚不清楚。此外， 技术实验室的变化，如样品采集、储存条件和移液技术，也可以 影响TL测量。这项研究的结果可能揭示出一种适合于大规模人群的最佳组织细胞 研究，澄清移液技术的精确度和其他技术实验室程序的研究 TL，并能够与使用不同测量方法得出的TL结果进行比较。 目标1将在5个不同的队列中，从出生到80岁，确定TL的可比性 取样组织[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]。AIM 2将检查技术 通过检测新鲜和存档的组织和DNA以及不同种类的组织和DNA进行TL测量的实验室差异 样品采集程序和移液技术(例如，手动与自动机器人)。AIM 3将测试 与不同组织和技术相关的潜在TL变异的关联性和估计效应大小 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，因为其严谨的设计将使人们能够对 从出生到80岁的TL的组织可比性，以及技术实验室的变异和估计的影响 使用多个qPCR方法的潜在TL变异的大小。端粒联盟的综合发现 将有助于形成TL测量的共识指导方针。这项研究具有创新性，因为它将是 首先，综合评估同一样本中的六个组织，跨越发育阶段和实验室 技术，并将为未来的比较奠定基础。由此生成的结果和原始数据 将与端粒研究网络联盟共享提案，以增加透明度、标准化 测量方法，并促进跨方法比较研究。

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.