Cancer Diagnosis Program | Biorepositories and Biospecimen Research Branch

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The microRNA spectrum in 12 body fluids.

Author(s): Weber JA, Baxter DH, Zhang S, Huang DY, Huang KH, Lee MJ, Galas DJ, Wang K

Publication: Clin Chem, 2010, Vol. 56, Page 1733-41

PubMed ID: 20847327 PubMed Review Paper? No

Purpose of Paper

Conclusion of Paper

Studies

1. Study Purpose

   The purpose of this study was to compare the miRNA profile of 12 different bodily fluids and to compare the miRNA profile of urine from healthy controls with that from patients with urogenital cancers and pregnant women. Breast milk, bronchial lavage, cerebrospinal fluid, peritoneal fluid, plasma, pleural fluid, saliva, seminal fluid, tears and urine were commercially obtained from five donors each (collection and processing details not specified). Colostrum from a single donor and amniotic fluid from one donor per trimester were also obtained from commercial sources.  For this study, urine was collected from three healthy patients, three with upper urinary tract urothelial cancer, three with bladder urothelial cancer and one specimen from a woman in each trimester of pregnancy. All specimens were centrifuged at 1000 g for 10 min. Fat was discarded from colostrum and breast milk. RNA was extracted from 300 µL of each specimen using the miRNeasy kit. RNA concentration was assessed by NanoDrop spectrophotometer. RNA integrity and concentration of small RNAs were evaluated using the RNA 6000 Pico chip on an Agilent bioanalyzer. RNA was reverse transcribed using the miScript Reverse Transcription kit. Levels of 714 miRNA were quantified by real-time PCR using the Human miScript Assay 384 set panel.

   Summary of Findings:

   Average RNA yields ranged from 113 µg/L to 48,240 µg/L among the bodily fluids examined, with the lowest median yields obtained from urine (94 µg/L), cerebrospinal fluid (111 µg/L), and plasma (308 µg/L) and the highest from seminal fluid (17,770 µg/L) and breast milk (47,240 µg/L). Similarly, the number of miRNAs detected (>80% global mean) varied among the fluids evaluated, with the fewest miRNAs detected in urine, pleural fluid and cerebrospinal fluid (204, 210 and 212 miRNAs, respectively) and the most detected in breast milk, seminal fluid and saliva (429, 436, and 458, respectively). Of the 714 miRNAs assessed, 600 were detected in one or more fluid and 61 were found in all of the fluids evaluated. Importantly, some miRNAs were specific to one fluid type (including miR-193b in breast milk, miR-224 in plasma, miR-508-5p in seminal fluid and miR-637 in tears), but no miRNAs specific to urine, bronchial lavage, or plural fluid were identified. Hierarchical clustering resulted in breast milk, seminal fluid, amniotic fluid, bronchial lavage, peritoneal fluid, saliva, and tears forming one cluster and plasma clustering separately.

   The number of detectable miRNAs in urine was dependent on patient condition. Generally, more miRNAs were detected in the urine of patients with cancer than in healthy controls.  Further the maternal urine specimen collected during the first trimester of pregnancy had more detectable miRNAs than those from healthy controls (336 versus a mean of 241 miRNAs). The authors identified miR-105*, miR- 449-3p, and miR-514 in urine from pregnant women and patients with cancer but not among healthy controls, while miR-620 and miR-125a-3p were only present in urine from cancer patients. Several miRNAs displayed different levels among cancer patients and healthy controls.

   Biospecimens

   • Bodily Fluid - Amniotic Fluid
   • Bodily Fluid - Bronchial Lavage
   • Bodily Fluid - Breast Milk
   • Bodily Fluid - Cerebrospinal Fluid
   • Bodily Fluid - Peritoneal Fluid
   • Bodily Fluid - Plasma
   • Bodily Fluid - Pleural Fluid
   • Bodily Fluid - Semen
   • Bodily Fluid - Urine
   • Bodily Fluid - Saliva
   • Bodily Fluid - Other
   • Bodily Fluid - Tears

   Preservative Types

   • None (Fresh)
   • Frozen

   Diagnoses:

   • Neoplastic - Carcinoma
   • Pregnant
   • Normal

   Platform:

   Analyte	Technology Platform
   RNA	Automated electrophoresis/Bioanalyzer
   RNA	Spectrophotometry
   RNA	Real-time qRT-PCR

   Pre-analytical Factors:

   Classification	Pre-analytical Factor	Value(s)
   Preaquisition	Diagnosis/ patient condition	Healthy First trimester of pregnancy Second trimester of pregnancy Third trimester of pregnancy Upper urinary tract urothelial cancer Bladder urothelial cancer
   Biospecimen Acquisition	Biospecimen location	Breast milk Bronchial lavage Cerebrospinal fluid Peritoneal fluid Plasma Pleural fluid Saliva Seminal fluid Tears Urine Colostrum

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