Human plasma and serum extracellular small RNA reference profiles and their clinical utility.
Human plasma and serum extracellular small RNA reference profiles and their clinical utility.
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DOI：
10.1073/pnas.1714397115
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发表时间：
2018-06-05
影响因子：
11.1
通讯作者：
Tuschl T
中科院分区：
文献类型：
--
作者：
Max KEA;Bertram K;Akat KM;Bogardus KA;Li J;Morozov P;Ben-Dov IZ;Li X;Weiss ZR;Azizian A;Sopeyin A;Diacovo TG;Adamidi C;Williams Z;Tuschl T
核酸介导遗传信息的储存和表达。细胞外DNA （exDNA）和exRNA是从细胞释放到细胞外环境的核酸的痕迹。它们作为疾病生物标志物的使用受到生物体液中大量RNA和dna降解酶导致的分离技术挑战的限制。使用专门为生物体液开发的分离方案，我们从13名健康志愿者中生成了血浆和血清exRNA参考谱，并确定了关键临床参数（如性别和禁食）的影响。令人惊讶的是，我们遇到了一位参与者，他的内分泌源exRNA贡献显著增加，稳定超过1年，在他的所有样本中都可以检测到，从而证明了这种方法的稳健性和循环rna作为生物标志物的临床潜力。循环细胞外rna （exrna）有潜力作为广泛的医疗条件的生物标志物。然而，现有的exRNA分离方法的局限性以及对影响人类样本中exRNA变异性的参数缺乏了解，可能会阻碍它们的成功发现和临床应用。使用变性剂、还原剂、蛋白水解和改进的有机萃取的组合，我们开发了一种自动化、高通量的方法，用于从同一生物流体样品中回收exrna和exDNA。我们应用该方法对13名健康志愿者在2个月期间的12个时间点收集的312份血浆和血清样本中的exrna进行了表征。小RNA cDNA文库测序发现，在女性中，上皮、肌肉和神经内分泌细胞特异性mirna增加了近两倍，而禁食和激素周期几乎没有影响。外部标准化有助于检测红细胞和血小板特异性miRNA贡献的定量差异以及生物液体之间miRNA浓度的差异。它还有助于确定具有独特exRNA表型的研究参与者，其miRNA特征为高达20倍的内分泌细胞特异性miRNA升高和两倍的总miRNA浓度升高，稳定超过1年。总的来说，这些结果证明了一种有效和定量的方法来识别exRNA表型，并表明血浆和血清RNA谱在几个月内是稳定的，可以在长期临床研究中进行常规监测。
Nucleic acids mediate storage and expression of genetic information. Extracellular DNA (exDNA) and exRNA are traces of nucleic acids released from cells into the extracellular environment. Their use as disease biomarkers has been limited by technical challenges in their isolation caused by abundant RNA- and DNA-degrading enzymes in biofluids. Using isolation protocols developed especially for biofluids, we generated plasma and serum exRNA reference profiles from 13 healthy volunteers over time and determined the effect of critical clinical parameters such as gender and fasting. Surprisingly, we encountered one participant with dramatically increased endocrine-origin exRNA contributions stable over 1 year and detectable in all of his samples, thereby demonstrating the robustness of this approach and the clinical potential of circulating RNAs as biomarkers. Circulating extracellular RNAs (exRNAs) have the potential to serve as biomarkers for a wide range of medical conditions. However, limitations in existing exRNA isolation methods and a lack of knowledge on parameters affecting exRNA variability in human samples may hinder their successful discovery and clinical implementation. Using combinations of denaturants, reducing agents, proteolysis, and revised organic extraction, we developed an automated, high-throughput approach for recovery of exRNAs and exDNA from the same biofluid sample. We applied this method to characterize exRNAs from 312 plasma and serum samples collected from 13 healthy volunteers at 12 time points over a 2-month period. Small RNA cDNA library sequencing identified nearly twofold increased epithelial-, muscle-, and neuroendocrine-cell–specific miRNAs in females, while fasting and hormonal cycle showed little effect. External standardization helped to detect quantitative differences in erythrocyte and platelet-specific miRNA contributions and in miRNA concentrations between biofluids. It also helped to identify a study participant with a unique exRNA phenotype featuring a miRNA signature of up to 20-fold elevated endocrine-cell–specific miRNAs and twofold elevated total miRNA concentrations stable for over 1 year. Collectively, these results demonstrate an efficient and quantitative method to discern exRNA phenotypes and suggest that plasma and serum RNA profiles are stable over months and can be routinely monitored in long-term clinical studies.
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影响因子：
3.7
作者：
Brown M;Suryawanshi H;Hafner M;Farazi TA;Tuschl T
通讯作者：
Tuschl T
影响因子：
5.8
作者：
Huntley, Melanie A.;Larson, Jessica L.;Kaminker, Joshua S.
通讯作者：
Kaminker, Joshua S.
影响因子：
6.4
作者：
Brase, Jan C.;Johannes, Marc;Sueltmann, Holger
通讯作者：
Sueltmann, Holger
影响因子：
20.3
作者：
Laffont, Benoit;Corduan, Aurelie;Provost, Patrick
通讯作者：
Provost, Patrick
影响因子：
2.7
作者：
Chomczynski, P;Mackey, K;Wilfinger, W
通讯作者：
Wilfinger, W