Identification of miR-31-5p, miR-141-3p, miR-200c-3p, and GLT1 as human liver aging markers sensitive to donor-recipient age-mismatch in transplants.
Identification of miR-31-5p, miR-141-3p, miR-200c-3p, and GLT1 as human liver aging markers sensitive to donor-recipient age-mismatch in transplants.
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DOI：
10.1111/acel.12549
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发表时间：
2017-04
期刊：
影响因子：
7.8
通讯作者：
Grazi GL
中科院分区：
文献类型：
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作者：
Capri M;Olivieri F;Lanzarini C;Remondini D;Borelli V;Lazzarini R;Graciotti L;Albertini MC;Bellavista E;Santoro A;Biondi F;Tagliafico E;Tenedini E;Morsiani C;Pizza G;Vasuri F;D'Errico A;Dazzi A;Pellegrini S;Magenta A;D'Agostino M;Capogrossi MC;Cescon M;Rippo MR;Procopio AD;Franceschi C;Grazi GL
为了解为何老年供体的肝脏能够成功用于移植，我们在71例移植前年龄为12 - 92岁的供体肝脏活检样本以及11例供体 - 受体年龄差异较大的移植后肝脏活检样本中寻找肝脏衰老的标志物。我们还评估了36例年龄不匹配受体的肝功能。主要发现如下：（i）通过对12例活检样本进行微小RNA（miR）和mRNA转录谱分析发现，miR - 31 - 5p、miR - 141 - 3p和miR - 200c - 3p随年龄增长而增加，并且通过实时定量聚合酶链反应（RT - qPCR）在总共58例活检样本中对结果进行了验证；（ii）通过qPCR对45个样本进行的端粒长度测量显示出明显的年龄相关性缩短；（iii）一种结合转录组和miR数据的生物信息学方法确定了可能的miR靶点，其中最具信息价值的是GLT1，一种在肝细胞中表达的谷氨酸转运体。通过荧光素酶实验证明GLT1是miR - 31 - 5p和miR - 200c - 3p的靶点，并且其mRNA（通过RT - qPCR检测）和蛋白质（通过免疫组织化学检测）在肝脏活检样本以及肝小叶中心区分别随年龄增长而显著降低；（iv）在11例供体 - 受体年龄差异极大的病例中发现，miR - 31 - 5p、miR - 141 - 3p和miR - 200c - 3p的表达受受体年龄（衰老环境）的显著影响；（v）通过N - 聚糖谱分析对受体血浆进行分析，该分析能够评估肝功能和生物学年龄，结果显示移植后肝功能恢复，与年龄不匹配无关，并且根据糖组学年龄，受体似乎“恢复年轻”。总之，我们确定了人类肝脏衰老的新标志物，它们在移植中供体 - 受体年龄不匹配方面的相关性，并为使用老年供体的器官提供了积极的证据。
To understand why livers from aged donors are successfully used for transplants, we looked for markers of liver aging in 71 biopsies from donors aged 12–92 years before transplants and in 11 biopsies after transplants with high donor–recipient age‐mismatch. We also assessed liver function in 36 age‐mismatched recipients. The major findings were the following: (i) miR‐31‐5p, miR‐141‐3p, and miR‐200c‐3p increased with age, as assessed by microRNAs (miRs) and mRNA transcript profiling in 12 biopsies and results were validated by RT–qPCR in a total of 58 biopsies; (ii) telomere length measured by qPCR in 45 samples showed a significant age‐dependent shortage; (iii) a bioinformatic approach combining transcriptome and miRs data identified putative miRs targets, the most informative being GLT1, a glutamate transporter expressed in hepatocytes. GLT1 was demonstrated by luciferase assay to be a target of miR‐31‐5p and miR‐200c‐3p, and both its mRNA (RT–qPCR) and protein (immunohistochemistry) significantly decreased with age in liver biopsies and in hepatic centrilobular zone, respectively; (iv) miR‐31‐5p, miR‐141‐3p and miR‐200c‐3p expression was significantly affected by recipient age (older environment) as assessed in eleven cases of donor–recipient extreme age‐mismatch; (v) the analysis of recipients plasma by N‐glycans profiling, capable of assessing liver functions and biological age, showed that liver function recovered after transplants, independently of age‐mismatch, and recipients apparently ‘rejuvenated’ according to their glycomic age. In conclusion, we identified new markers of aging in human liver, their relevance in donor–recipient age‐mismatches in transplantation, and offered positive evidence for the use of organs from old donors.