Characterization of Serum Extracellular Vesicles with Human Age

我们继续对来自基于人群的纵向研究“LEPS生命周期中多样性社区的健康老龄化”（HANDLS）的选定年龄组参与者的EV属性进行表征。我们使用横断面和纵向方法分析了来自HANDLS研究的循环血浆EV，以解决社区居民个体的年龄相关变化。今年，我们扩展了EV工作，以进一步探索脆弱性。我们研究了EV中的线粒体DNA（mtDNA）和炎性蛋白是否可能在虚弱中充当损伤相关分子模式（DAMP）分子。为了解决EV及其相关的mtDNA和炎症蛋白货物是否随虚弱而改变，EV从非虚弱（n = 90）和虚弱（n = 87）的中年（45-55岁）参与者中分离，这些参与者来自整个生命周期（HANDLS）研究的多样性社区健康老龄化。EV浓度在虚弱的白色参与者中最高。EV mtDNA水平显着高于体弱的人相比，非体弱的人。EV中六种炎性蛋白（FGF-21，HGF，IL-12 B，PD-L1，PRDX 3和STAMBP）的存在与虚弱显著相关。EV炎性蛋白质显著改变的脆弱状态，种族，性别和贫困状况。值得注意的是，与虚弱和非虚弱的非裔美国人参与者相比，虚弱的白色参与者具有更高水平的EV相关CD 5、CD 8A、CD 244、CXCL 1、CXCL 6、CXCL 11、LAP-TGF-β-1和MCP-4。生活在贫困线以下的虚弱白色参与者具有更高水平的EV相关uPA。EV相关的CCL 28水平在非虚弱女性中最高，CXCL 1在非虚弱男性中最高。生活在贫困以下的男性有较高水平的CD 5，CD 8A，CXCL 1，LAP-TGF-beta-1和uPA。生活在贫困以上的人的CXCL 6水平显着更高。EV mtDNA水平与炎性蛋白的存在有显著相关性。这些数据表明，线粒体DNA的EV可能作为一个DAMP分子在脆弱。它与趋化因子和其他炎性EV货物蛋白的相关性可能有助于脆弱表型。此外，健康的社会决定因素，贫困，影响中年EV的炎症货物。我们还研究了死亡率。即使在COVID-19大流行之前，美国预期寿命的下降也归因于不同种族和民族中年人死亡率的增加，这表明需要标记物来识别有早期死亡风险的个体。细胞外囊泡（EV）是能够穿梭功能性蛋白质、核酸和脂质的小的脂质结合囊泡。鉴于它们作为细胞间通讯者和疾病的潜在生物标志物的作用，我们探讨了循环EV是否可能是前瞻性，种族和社会经济多样性中年队列中死亡率的标志物。我们从76名在5年内死亡的个体（平均年龄= 59.6岁）和76名年龄、种族和贫困状况相匹配的存活个体中分离出血浆EV。EV浓度、大小或EV相关线粒体DNA水平与死亡率无显著差异。我们发现，包括CCL 23、CSF-1、CXCL 9、GDNF、MCP-1、STAMBP和4 E-BP 1在内的几种EV相关炎症蛋白与死亡率显著相关。IL-10 RB和CDCP 1更可能存在于来自死亡个体的血浆EV中，而不是其活着的对应物中。我们还报告了EV相关炎症蛋白与贫困状况，种族和性别的差异。我们的研究结果表明，血浆EV相关炎症蛋白是有前途的潜在临床死亡率的生物标志物。

We continue to characterize EV attributes across selected age groups of participants from the population based, longitudinal study, Healthy Aging in Neighborhoods of Diversity across the Life Span (HANDLS) in LEPS. We analyzed circulating plasma EVs from the HANDLS study using both a cross-sectional and longitudinal approach to address age-related changes in community-dwelling individuals. This year we extended our EV work to further explore frailty. We examined whether mitochondrial DNA (mtDNA) and inflammatory proteins in EVs may act as damage-associated molecular pattern (DAMP) molecules in frailty. To address whether EVs and their associated mtDNA and inflammatory protein cargo are altered with frailty, EVs were isolated from non-frail (n = 90) and frail (n = 87) middle-aged (45-55 years) participants from the Healthy Aging in Neighborhoods of Diversity across the Life Span (HANDLS) study. EV concentration was highest in frail White participants. EV mtDNA levels were significantly higher in frail individuals compared to non-frail individuals. The presence of six inflammatory proteins in EVs (FGF-21, HGF, IL-12B, PD-L1, PRDX3, and STAMBP) were significantly associated with frailty. EV inflammatory proteins were significantly altered by frailty status, race, sex, and poverty status. Notably, frail White participants had higher levels of EV-associated CD5, CD8A, CD244, CXCL1, CXCL6, CXCL11, LAP-TGF-beta-1 and MCP-4 compared to frail and non-frail African American participants. Frail White participants living below poverty had higher levels of EV-associated uPA. EV-associated CCL28 levels were highest in non-frail women and CXCL1 were highest in non-frail men. Men living below poverty had higher levels of CD5, CD8A, CXCL1, LAP-TGF-beta-1, and uPA. CXCL6 levels were significantly higher in individuals living above poverty. There was a significant correlation between EV mtDNA levels and the presence of inflammatory proteins. These data suggest that mtDNA within EVs may act as a DAMP molecule in frailty. Its association with chemokines and other inflammatory EV cargo proteins, may contribute to the frailty phenotype. In addition, the social determinant of health, poverty, influences the inflammatory cargo of EVs in midlife. We also examined mortality. Even before the COVID-19 pandemic declines in life expectancy in the United States were attributed to increased mortality rates in midlife adults across racial and ethnic groups, indicating a need for markers to identify individuals at risk for early mortality. Extracellular vesicles (EVs) are small, lipid-bound vesicles capable of shuttling functional proteins, nucleic acids, and lipids. Given their role as intercellular communicators and potential biomarkers of disease, we explored whether circulating EVs may be markers of mortality in a prospective, racially, and socioeconomically diverse middle-aged cohort. We isolated plasma EVs from 76 individuals (mean age = 59.6 years) who died within a 5 year period and 76 surviving individuals matched by age, race, and poverty status. There were no significant differences in EV concentration, size, or EV-associated mitochondrial DNA levels associated with mortality. We found that several EV-associated inflammatory proteins including CCL23, CSF-1, CXCL9, GDNF, MCP-1, STAMBP, and 4E-BP1 were significantly associated with mortality. IL-10RB and CDCP1 were more likely to be present in plasma EVs from deceased individuals than in their alive counterparts. We also report differences in EV-associated inflammatory proteins with poverty status, race, and sex. Our results suggest that plasma EV-associated inflammatory proteins are promising potential clinical biomarkers of mortality.