Circulating microRNAs in patients with coronary artery disease.
Author(s): Fichtlscherer S, De Rosa S, Fox H, Schwietz T, Fischer A, Liebetrau C, Weber M, Hamm CW, Röxe T, Müller-Ardogan M, Bonauer A, Zeiher AM, Dimmeler S
Publication: Circ Res, 2010, Vol. 107, Page 677-84
PubMed ID: 20595655 PubMed Review Paper? No
Purpose of Paper
The purpose of this paper was to compare extraction methods and investigate the effects of patient gender and age; levels of creatinine and urea; current use of statins, ACE/AR‐ blockers, or aspirin; and diagnosis of diabetes on levels of microRNA (miRNA, miR) in plasma and serum.
Conclusion of Paper
Levels of miR-92a were higher in plasma and serum when extraction was with the miRNeasy Kit rather than Trizol alone or in combination with isopropanol or ammonium chloride precipitation. Plasma had higher levels of miR-92a than serum, regardless of extraction method, and the authors report that EDTA plasma was superior to citrate plasma. Levels of individual miRNAs were negatively correlated with patient age; levels of creatinine and urea; current use of ACE/AR‐ blockers, statins, and aspirin; as well as diagnosis of diabetes and were positively correlated with total, high-density lipoprotein (HDL), and low-density lipoprotein (LDL) cholesterol levels.
Studies
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Study Purpose
The purpose of this study was to compare extraction methods and investigate the effects of patient gender and age; levels of creatinine and urea; current use of statins, ACE/AR‐ blockers, or aspirin; and diagnosis of diabetes on levels of miRNA in plasma and serum. Blood was collected from 17 healthy volunteers and 36 patients with coronary artery disease into EDTA tubes and serum tubes. Details of blood processing to plasma and serum and subsequent storage were not specified. To determine the best extraction method, RNA was extracted from the serum and EDTA plasma of three healthy patients using miRNeasy, Trizol, Trizol with isopropanol precipitation, and Trizol with ammonium chloride precipitation. Selected miRNAs were quantified by TaqMan RT-PCR kits.
Summary of Findings:
Levels of miR-92a were higher in plasma and serum when extraction was with the miRNeasy Kit rather than Trizol alone or in combination with isopropanol or ammonium chloride precipitation. The authors report similar results for miR-126. Plasma had higher levels of miR-92a than serum, regardless of extraction method. The authors report that EDTA plasma was superior to citrate plasma. miR-155 levels were weakly to modestly negatively correlated with patient age (R=-0.395, P=0.003); levels of creatinine (R=-0.390, P=0.19)and urea (-0.393, P=0.018); current use of statins (R=-0.528, P<0.001), ACE/AR‐ blockers (R=-0.331, P=0.015), and aspirin (R=-0.393, P<0.001); and diagnosis with diabetes (R=-0.315, P=0.021) and were positively correlated with total (R=0.363, P=0.008), HDL (R=0.376, P=0.006), and LDL (0.325, P=0.021) cholesterol levels. All of the aforementioned variables with the exception of patient age, gender, and total and LDL cholesterol levels, were correlated with levels of one or more of the selected miRNAs (miR-126, miR-17, miR-92a, miR-199a, miR-145, miR-133a, and/or miR-208a).
Biospecimens
Preservative Types
- Frozen
Diagnoses:
- Coronary Artery Disease
- Diabetes Type 2
- Normal
Platform:
Analyte Technology Platform RNA Real-time qRT-PCR Pre-analytical Factors:
Classification Pre-analytical Factor Value(s) Preaquisition Diagnosis/ patient condition CAD and diabetes
CAD alone
Healthy
Preaquisition Patient gender Female
Male
Preaquisition Patient age 23-40 years
56-78 years
Real-time qRT-PCR Specific Targeted nucleic acid miR-126
miR-17
miR-92a
miR-155
miR-199a
miR-145
miR-133a
miR-208a
Biospecimen Aliquots and Components Blood and blood products Plasma
Serum
Analyte Extraction and Purification Analyte isolation method miRNeasy
Trizol
Trizol with isopropanol precipitation
Trizol with ammonium chloride precipitation
Biospecimen Acquisition Anticoagulant Citrate
EDTA
None