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

项目摘要 端粒长度（TL）已被确立为细胞衰老的标志，并且是细胞衰老的标志之一。 衰老研究表明，年龄相关性TL与预测疾病发病率的广泛风险因素有关 早期死亡率。对端粒科学感兴趣的研究人员仍然担心精确度 测量和再现性，这是该领域和更广泛的科学中的一个紧迫问题。 一个关键问题是测量来自大群体不同类型组织细胞的TL- 基于研究。血液通常用于TL研究。然而，血液采集是相对侵入性的， 成本高，并且在收集地点和收集人方面受到限制。科学进步的理解 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。 TL如何在普通组织细胞和发育阶段中在个体内和个体间存在差异， T/S比、单色多重和绝对定量PCR（qPCR）方法未知。此外，委员会认为， 技术实验室变化，例如样品收集、储存条件和移液技术，也可 影响TL测量。这项研究的结果可能揭示了一个最佳的组织细胞，为大人口为基础的 研究，澄清移液技术和其他技术实验室程序的精度， 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将确定从出生到80岁的5个不同队列中， 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]。目标2将审查技术 通过测试新鲜与存档组织和DNA，以及各种 样品收集程序和移液技术（即，手动与自动机器人）。目标3将测试 与不同组织和技术相关的潜在TL变异的关联和估计效应量 涉及多系统生物学和表型测量的实验室技术。 这项研究是重要的，因为它的严格设计将使强有力的推论的影响， TL从出生到80岁的组织可比性，以及技术实验室变化和估计影响 使用多个qPCR方法来确定潜在TL变异的大小。端粒联盟的综合研究结果 将有助于为TL测量产生共识指南。这项研究是创新的，因为它将是 第一次全面评估来自同一样本的六种组织，跨越发育阶段和实验室 技术，并将为未来的比较奠定基础。由此产生的结果和原始数据 该提案将与端粒研究网络联盟分享，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.