Ben-Dov Iddo Z, Tan Ying-Cai, Morozov Pavel, Wilson Patricia D, Rennert Hanna, Blumenfeld Jon D, Tuschl Thomas
Laboratory of RNA Molecular Biology, Howard Hughes Medical Institute, The Rockefeller University, New York, New York, United States of America.
Molecular Pathology Laboratory, New York Presbyterian Hospital, Cornell University, New York, New York, United States of America ; Pathology and Laboratory Medicine, Weill Medical College, Cornell University, New York, New York, United States of America.
PLoS One. 2014 Jan 29;9(1):e86856. doi: 10.1371/journal.pone.0086856. eCollection 2014.
Autosomal dominant polycystic kidney disease (ADPKD) is clinically heterogenic. Biomarkers are needed to predict prognosis and guide management. We aimed to profile microRNA (miRNA) in ADPKD to gain molecular insight and evaluate biomarker potential.
Small-RNA libraries were generated from urine specimens of ADPKD patients (N = 20) and patients with chronic kidney disease of other etiologies (CKD, N = 20). In this report, we describe the miRNA profiles and baseline characteristics. For reference, we also examined the miRNA transcriptome in primary cultures of ADPKD cyst epithelia (N = 10), normal adult tubule (N = 8) and fetal tubule (N = 7) epithelia.
In primary cultures of ADPKD kidney cells, miRNA cistrons mir-143(2) (9.2-fold), let-7i(1) (2.3-fold) and mir-3619(1) (12.1-fold) were significantly elevated compared to normal tubule epithelia, whereas mir-1(4) members (19.7-fold), mir-133b(2) (21.1-fold) and mir-205(1) (3.0-fold) were downregulated (P<0.01). Expression of the dysregulated miRNA in fetal tubule epithelia resembled ADPKD better than normal adult cells, except let-7i, which was lower in fetal cells. In patient biofluid specimens, mir-143(2) members were 2.9-fold higher in urine cells from ADPKD compared to other CKD patients, while expression levels of mir-133b(2) (4.9-fold) and mir-1(4) (4.4-fold) were lower in ADPKD. We also noted increased abundance mir-223(1) (5.6-fold), mir-199a(3) (1.4-fold) and mir-199b(1) (1.8-fold) (P<0.01) in ADPKD urine cells. In ADPKD urine microvesicles, miR-1(2) (7.2-fold) and miR-133a(2) (11.8-fold) were less abundant compared to other CKD patients (P<0.01).
We found that in ADPKD urine specimens, miRNA previously implicated as kidney tumor suppressors (miR-1 and miR-133), as well as miRNA of presumed inflammatory and fibroblast cell origin (miR-223/miR-199), are dysregulated when compared to other CKD patients. Concordant with findings in the primary tubule epithelial cell model, this suggests roles for dysregulated miRNA in ADPKD pathogenesis and potential use as biomarkers. We intend to assess prognostic potential of miRNA in a followup analysis.
常染色体显性遗传性多囊肾病（ADPKD）在临床上具有异质性。需要生物标志物来预测预后并指导治疗。我们旨在分析ADPKD中的微小RNA（miRNA），以获得分子层面的认识并评估其作为生物标志物的潜力。
从ADPKD患者（N = 20）和其他病因的慢性肾脏病患者（CKD，N = 20）的尿液标本中构建小RNA文库。在本报告中，我们描述了miRNA谱和基线特征。作为对照，我们还检测了ADPKD囊肿上皮细胞（N = 10）、正常成人肾小管（N = 8）和胎儿肾小管上皮细胞（N = 7）原代培养物中的miRNA转录组。
在ADPKD肾细胞原代培养物中，与正常肾小管上皮相比，miRNA顺反子mir - 143(2)（9.2倍）、let - 7i(1)（2.3倍）和mir - 3619(1)（12.1倍）显著上调，而mir - 1(4)成员（19.7倍）、mir - 133b(2)（21.1倍）和mir - 205(1)（3.0倍）下调（P<0.01）。除了let - 7i在胎儿细胞中较低外，胎儿肾小管上皮中失调的miRNA表达与ADPKD的相似性高于正常成人细胞。在患者生物流体标本中，与其他CKD患者相比，ADPKD患者尿液细胞中mir - 143(2)成员高2.9倍，而mir - 133b(2)（4.9倍）和mir - 1(4)（4.4倍）在ADPKD中的表达水平较低。我们还注意到ADPKD尿液细胞中mir - 223(1)（5.6倍）、mir - 199a(3)（1.4倍）和mir - 199b(1)（1.8倍）丰度增加（P<0.01）。与其他CKD患者相比，ADPKD尿液微泡中的miR - 1(2)（7.2倍）和miR - 133a(2)（11.8倍）含量较少（P<0.01）。
我们发现，在ADPKD尿液标本中，与其他CKD患者相比，先前被认为是肾肿瘤抑制因子的miRNA（miR - 1和miR - 133）以及推测来源于炎症和成纤维细胞的miRNA（miR - 223/miR - 199）失调。与原代肾小管上皮细胞模型中的发现一致，这表明失调的miRNA在ADPKD发病机制中起作用，并有可能用作生物标志物。我们打算在后续分析中评估miRNA的预后潜力。
