Heparin Selectively Affects the Quantification of MicroRNAs in Human Blood Samples

Boeckel, Jes‐Niels; Thome, Claudia; Leistner, David M.; Zeiher, Andreas M.; Fichtlscherer, Stephan; Dimmeler, Stefanie

doi:10.1373/clinchem.2012.199505

Cited by 90 publications

(60 citation statements)

References 5 publications

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“…We observed that heparin did not affect the quantification of lncRNAs. This apparent discrepancy with the study by Boeckel et al 40 can have several explanations. First, the concentration of heparin in patient blood (1 IU/mL of blood) is lower than in heparinized blood collection tubes (11-32 IU/mL).…”

Section: Downloaded Fromcontrasting

confidence: 75%

“…Boeckel et al 40 reported that heparin affects the quantification of microRNAs by polymerase chain reaction in human blood samples. Because all patients with MI enrolled in the study received a bolus of 5000 IU of heparin, we tested a potential confounding effect of heparin on levels of lncRNAs.…”

Section: Downloaded Frommentioning

confidence: 99%

“…First, the concentration of heparin in patient blood (1 IU/mL of blood) is lower than in heparinized blood collection tubes (11-32 IU/mL). Second, RNA preparation from PAXgene tubes does not require a phenol-chloroform extraction step, which has been performed to measure microRNAs in the study by Boeckel et al, 40 and which copurifies nucleic acids with heparin. 41 Third, we have measured lncRNAs, whereas Boeckel et al 40 have measured microRNAs, which may be more sensitive to interference by heparin than lncRNAs.…”

Section: Downloaded Frommentioning

confidence: 99%

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Long Noncoding RNAs in Patients With Acute Myocardial Infarction

Vausort

Wagner²,

Devaux

2014

Circulation Research

452

352

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Rationale: Long noncoding RNAs (lncRNAs) constitute a novel class of noncoding RNAs that regulate gene expression. Although recent data suggest that lncRNAs may be associated with cardiac disease, little is known about lncRNAs in the setting of myocardial ischemia. Objective: To measure lncRNAs in patients with myocardial infarction (MI). Methods and Results: We enrolled 414 patients with acute MI treated by primary percutaneous coronary intervention. Blood samples were harvested at the time of reperfusion. Expression levels of 5 lncRNAs were measured in peripheral blood cells by quantitative polymerase chain reaction: hypoxia inducible factor 1A antisense RNA 2, cyclin-dependent kinase inhibitor 2B antisense RNA 1 (ANRIL), potassium voltage-gated channel, KQT-like subfamily, member 1 opposite strand/antisense transcript 1 (KCNQ1OT1), myocardial infarction–associated transcript, and metastasis-associated lung adenocarcinoma transcript 1. Levels of hypoxia inducible factor 1A antisense RNA 2, KCNQ1OT1, and metastasis-associated lung adenocarcinoma transcript 1 were higher in patients with MI than in healthy volunteers ( P <0.01), and levels of ANRIL were lower in patients with MI ( P =0.003). Patients with ST-segment–elevation MI had lower levels of ANRIL ( P <0.001), KCNQ1OT1 ( P <0.001), myocardial infarction–associated transcript ( P <0.001), and metastasis-associated lung adenocarcinoma transcript 1 ( P =0.005) when compared with patients with non–ST-segment–elevation MI. Levels of ANRIL were associated with age, diabetes mellitus, and hypertension. Patients presenting within 3 hours of chest pain onset had elevated levels of hypoxia inducible factor 1A antisense RNA 2 when compared with patients presenting later on. ANRIL, KCNQ1OT1, myocardial infarction–associated transcript, and metastasis-associated lung adenocarcinoma transcript 1 were significant univariable predictors of left ventricular dysfunction as assessed by an ejection fraction ≤40% at 4-month follow-up. In multivariable and reclassification analyses, ANRIL and KCNQ1OT1 improved the prediction of left ventricular dysfunction by a model, including demographic features, clinical parameters, and cardiac biomarkers. Conclusions: Levels of lncRNAs in blood cells are regulated after MI and may help in prediction of outcome. This motivates further investigation of the role of lncRNAs after MI.

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confidence: 75%

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confidence: 99%

Section: Downloaded Frommentioning

confidence: 99%

See 1 more Smart Citation

Long Noncoding RNAs in Patients With Acute Myocardial Infarction

Vausort

Wagner²,

Devaux

2014

Circulation Research

452

352

View full text Add to dashboard Cite

show abstract

“…Boeckel et al [31] indicated that heparin can inhibit RNA quantification in vitro by interfering with the DNA polymerase used in the qPCR reaction. In the present study, we detected the expression of Homer in peripheral blood leukocytes added with heparin to assess the stability of Homer in IS patients.…”

Section: Discussionmentioning

confidence: 99%

Diagnostic Potential of Differentially Expressed Homer1 and Homer2 in Ischemic Stroke

Zhu¹,

Zuo²,

Ji³

et al. 2016

Cell Physiol Biochem

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Background: Ischemic stroke (IS) is an extremely heterogeneous disease with variable pathogenesis. Due to the lack of early diagnostic marker, the mortality rate of IS remains high worldwide. The family of Homer plays an important role in the pathology of atherosclerotic plaque. In this study, we have investigated its expression pattern and clinical significance in IS. Methods: RT-qPCR was performed to detect the expression of Homer1, Homer2, and Homer3. Results: We found that the mRNA levels of Homer1 (p<0.001) and Homer2 (p<0.001), but not Homer3, in large-artery atherosclerosis (LAA) strokes were significantly upregulated than those in non-LAA strokes and controls. Multinomial logistic regression analyses showed that, although none of the Homer was associated with non-LAA strokes, higher Homer1 (adjusted OR=1.337, 95% CI: 1.227-1.458) and Homer2 (adjusted OR=1.099, 95% CI: 1.062-1.138) levels showed significant associations with increased odds of having LAA stroke, compared with the controls. The receiver operating characteristic (ROC) curves showed that the combination of Homer1 and Homer2 had a better diagnostic accuracy to differentiate LAA strokes from non-LAA strokes and controls, and the sensitivity and specificity ratios were 80.5%/90.4% and 98.0%/70.3%, respectively. Conclusion: Our data suggested that Homer1 and Homer2 might be considered as novel diagnostic biomarkers for LAA stroke.

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“…It is also vital to take into account the type of biological fluid used to measure miRNA concentration, as it has been shown that higher miRNA concentrations were measured in serum compared with plasma samples from the same individual [80]. Some studies in patients with coronary artery disease have shown that administration of heparin interferes with miRNA quantification, and that use of ACE inhibitors may also influence miRNA levels [81,82]. Renal function has also been found to influence plasma levels of miRNA [83].…”

Section: Limitations Of Mirnas As Biomarkersmentioning

confidence: 99%

MicroRNAs as Potential Prognosticators of Neurological Outcome in Out-Of-Hospital Cardiac Arrest Patients

et al. 2017

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Out-of-hospital cardiac arrest survival rates have increased due to advancement in resuscitative measures, yet approximately 90% of survivors ultimately die or have severe neurologic dysfunction caused by ischemic injury. Currently, there are few early prognostic indicators of which patients have possibility of meaningful recovery. This leads to uncertainty for families and clinicians, as well as aggressive, invasive and expensive treatments despite medical futility. Several biomarkers investigated in traumatic brain injury have shown prognostication potential in ischemic brain injury. miRNAs, small noncoding RNAs responsible for gene regulation, have been studied in cardiovascular diseases, and have shown prognostication potential due to tissue specificity and stability in circulation. This review discusses available evidence on miRNAs prognosticating neurological outcomes after out-of-hospital cardiac arrest.

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Heparin Selectively Affects the Quantification of MicroRNAs in Human Blood Samples

Cited by 90 publications

References 5 publications

Long Noncoding RNAs in Patients With Acute Myocardial Infarction

Long Noncoding RNAs in Patients With Acute Myocardial Infarction

Diagnostic Potential of Differentially Expressed Homer1 and Homer2 in Ischemic Stroke

MicroRNAs as Potential Prognosticators of Neurological Outcome in Out-Of-Hospital Cardiac Arrest Patients

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