Inclusive Transcriptomics to Uncover Molecular Mechanisms Underlying Healthspan

项目摘要 当前老年科学的一个主要目标是通过在老年时保持生理功能来延长健康寿命。 如NIA PA-17-089中所强调的，使用次要分析 现有的生物标本和数据来测试这一领域的新假设。下一代测序方法 例如转录组学在这方面具有很大前景，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 忽视为了解决这个问题，我们使用来自人类受试者的现有样本进行了初步研究， 总转录组随年龄增长和运动而变化--少数几个被证实的保存 生理功能随着年龄的增长我们发现与年龄相关的炎症相关基因转录增加， 衰老的一个主要标志，减少健康寿命，以及非编码RNA（ncRNA）的主要变化。在 特别是，我们观察到：1）重复元件（RE）转录本的年龄相关性增加，通常 被忽视的ncRNA来源于重复序列，占基因组的50%以上; 2）强相关性 RE转录水平与炎症和生理功能之间的关系;以及3）RE转录水平与炎症和生理功能之间的显著逆转。 这些事件与锻炼。这些新的观察结果表明，RE转录本的失调可能反映了 一种以前未被认识到的临床相关衰老机制。因此，我们建议延长这项 初步研究了更大的现有样本和数据集，我们假设： ·衰老将与RE/其他ncRNA的增加和基因表达变化相关联， 具有老化的标志（例如，炎症），这将与运动逆转。为了验证这一点，我们将分离 我们已经从仔细匹配的组（10名受试者）中收集了来自外周血细胞的RNA 每组）的年轻人，老年人和老年人锻炼的成年人，以及老年人谁参加了一个 运动干预，我们将通过总RNA-seq表征转录组。 · RE/其他ncRNA将与炎症和生理功能随衰老的差异相关，以及 因此，可以提供对健康寿命的新机制的深入了解。我们将RE/ncRNA与基因 炎症循环标志物的表达和一系列生理功能测量 （血管，运动/身体，认知和代谢）在更大的老年人组（n = 110）。 ·RE/ncRNA/基因表达随衰老和运动的变化可以解释为： 染色质结构，这可以为未来的研究和干预提供新的目标。到 为了测试这一点，我们将使用ATAC-seq分析上述样品中的全局染色质可及性。 这将是第一项研究，以描绘总转录组的变化与衰老和运动， 将它们与炎症和生理功能的多个领域联系起来， 在人类受试者中表征全基因组染色质随衰老/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.