Inclusive Transcriptomics to Uncover Molecular Mechanisms Underlying Healthspan

项目总结 当前老年学的一个主要目标是通过保护老年人的生理功能来增加健康寿命 年龄，而且正如在NIA PA-17-089中强调的那样，使用二次分析对 现有的生物标本和数据来检验这一领域的新假设。下一代测序方法 例如转录组在这方面很有希望，但大多数关于衰老的测序数据都是基于 已知的蛋白质/基因，而非编码遗传物质(基因组的大部分)的作用往往是 被忽视了。为了解决这一问题，我们使用来自人类受试者的现有样本进行了初步研究 总转录组随年龄和运动的变化--为数不多的被证实的保存策略之一 生理机能随年龄的变化。我们发现与年龄相关的与炎症相关的基因转录增加， 衰老的中心特征是缩短健康寿命，以及非编码RNA(NcRNA)的重大变化。在……里面 特别是，我们观察到：1)与年龄相关的重复元件(RE)转录本的增加，通常 被忽视的来自重复序列的ncRNA，这些重复序列构成了基因组的50%；2)强相关性 在RE转录本水平与炎症和生理功能之间；以及3)显著逆转 这些活动都需要锻炼。这些新的观察结果表明，RE转录本的失调可能反映了 一种先前未被认识的、临床上相关的衰老机制。因此，我们建议延长这一期限 对一组更大的现有样本和数据进行的初步研究，我们假设： ·衰老将与RE/其他ncRNAs增加和基因表达变化一致相关 有衰老的特征(例如，炎症)，这将通过锻炼而逆转。为了测试这一点，我们将分离 我们已经从精心配对的组(10名受试者)收集的外周血细胞RNA 每组)，包括年轻的、年长的和年长的锻炼的成年人，以及参加 运动干预，我们将通过总RNA-seq来表征转录组。 ·RE/其他ncRNA将与随年龄增长的炎症和生理功能的差异有关，以及 因此，可能会提供对健康寿命的新机制的洞察。我们将RE/ncRNA与基因联系起来 循环炎症标志物的表达及一系列生理功能检测 (血管、运动/身体、认知和代谢)在更大的老年人群体(n=110)中。 ·RE/ncRNA/基因表达随年龄和运动的变化可能可以用不同的 染色质结构，这可能为未来的研究和干预提供新的靶点。至 测试这一点，我们将使用atac-seq在上面描述的样本中分析全局染色质的可及性。 这将是同类研究中第一次描述总转录组随年龄和运动的变化，以及 将它们与炎症和生理功能的多个领域联系起来，这是任何类型的第一次研究 描述人类受试者全基因组染色质随年龄/运动的变化。

PROJECT SUMMARY A major goal of current geroscience is to increase healthspan by preserving physiological function into older age, and as emphasized in NIA PA-17-089, there is considerable potential in using secondary analyses of existing biospecimens and data to test new hypotheses in this area. Next-generation sequencing approaches such as transcriptomics hold great promise in this context, but most sequencing data on aging are based on known proteins/genes, while the role of non-coding genetic material (the majority of the genome) is often overlooked. To address this, we used existing samples from human subjects to perform a preliminary study of total transcriptome changes with aging and exercise—one of the few proven strategies for preserving physiological function with age. We found age-related increases in gene transcription related to inflammation, a central hallmark of aging that reduces healthspan, and major changes in non-coding RNA (ncRNA). In particular, we observed: 1) an age-related increase in repetitive element (RE) transcripts, commonly overlooked ncRNA derived from repeat sequences that make up > 50% of the genome; 2) strong correlations between RE transcript levels and both inflammation and physiological function; and 3) a marked reversal of these events with exercise. These novel observations suggest that dysregulation of RE transcripts may reflect a previously unrecognized, clinically relevant mechanism of aging. Therefore, we propose to extend this preliminary study to a larger set of existing samples and data, and we hypothesize that: · Aging will be associated with an increase in RE/other ncRNAs and gene expression changes consistent with hallmarks of aging (e.g., inflammation), which will be reversed with exercise. To test this, we will isolate RNA from peripheral blood cells that we have already collected from carefully matched groups (10 subjects per group) of young, older, and older exercising adults, as well as older adults who participated in an exercise intervention, and we will characterize the transcriptome by total RNA-seq. · RE/other ncRNAs will be related to differences in inflammation and physiological function with aging, and may, therefore, provide insight into novel mechanisms of healthspan. We will relate RE/ncRNA and gene expression to circulating markers of inflammation and a battery of physiological function measurements (vascular, motor/physical, cognitive and metabolic) in a larger group of older adults (n = 110). · Changes in RE/ncRNA/gene expression with aging and exercise may be explained by differences in chromatin structure, which could provide insight into novel targets for future studies and interventions. To test this, we will profile global chromatin accessibility in samples described above using ATAC-seq. This will be the first research of its kind to profile total transcriptome changes with aging and exercise and to relate them to inflammation and multiple domains of physiological function, and the first research of any kind to characterize genome-wide chromatin changes with aging/exercise in human subjects.