Heparinase treatment of heparin-contaminated plasma from coronary artery bypass grafting patients enables reliable quantification of microRNAs

Кондратов, К. А.; Kurapeev, Dmitry; Попов, М. А.; Сидорова, М В; Минасян, С. М.; Galagudza, M. M.; Kostareva, Anna; Федоров, А. В.

doi:10.1016/j.bdq.2016.03.001

Cited by 30 publications

(15 citation statements)

References 19 publications

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“…16 In the setting of a brain tumor, cfDNA is difficult to detect in plasma DNA due to the blood-brain barrier, 27 which is consistent with our results where EGFRvIIIderived PCR amplicons were not obtained from either cfDNA or evDNA derived from plasma, even with modifications to enhance sensitivity, such as preamplification 13 and heparinase I treatment. 14 In contrast, a previous study analyzing patients with brain tumors, including 8 GBMs, shows that tumor DNA was detectable in CSF, 28 as in one of the cases in the current study, suggesting that this source of cfDNA provides a means to detect EGFRvIII mutations based on a tumor-defined, patient-specific breakpoint fingerprint.…”

Section: Neuro-oncologycontrasting

confidence: 82%

Mapping of genomic EGFRvIII deletions in glioblastoma: insight into rearrangement mechanisms and biomarker development

Koga

Figueroa

et al. 2018

Neuro-Oncology

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Section: Neuro-oncologycontrasting

confidence: 82%

Mapping of genomic EGFRvIII deletions in glioblastoma: insight into rearrangement mechanisms and biomarker development

Koga

Figueroa

et al. 2018

Neuro-Oncology

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“…Kondratov et al . recently reported the use of calibration curves to determine amplification efficiencies and the presence of inhibitors in heparinase I treated miRNA samples from CABG patients and successful quantification of miR-1-3p and miR-208a by using a heparinase I treatment protocol for heparinized plasma quite similar to ours 41 . However, in our study, we developed a simple protocol with spike-in C. elegans cel-miR-54 as control, in which RNA samples treated with heparinase I could be directly used in reverse transcription.…”

Section: Discussionmentioning

confidence: 78%

Establishment of an easy and straight forward heparinase protocol to analyse circulating and myocardial tissue micro-RNA during coronary artery-bypass-graft surgery

Engler

Dreja

et al. 2018

Sci Rep

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Coronary artery-bypass-graft (CABG) surgery is associated with myocardial damage and increased blood concentrations of circulating microRNAs (miRNA). However, whether and to what extent these miRNAs relate to cardiac tissue miRNA expression have not yet been explored. Since plasma miRNA quantification in samples from cardiopulmonary bypass (CPB) patients is severely hampered by heparin, we established and validated successfully a protocol to reliably measure miRNA in 49 heparinized patients undergoing CABG so as to investigate the relationship between circulating and right atrial miRNAs. Plasma and right atrial expression of miR-1, miR-133a, miR-423-5p, and miR-499 were measured before and after CPB, as well as miRNAs in plasma 24 h thereafter. All plasma miRNAs increased significantly with surgery while cardiac tissue expression of only miR-133a (1.4-fold; p = 0.003) and miR-423-5p (1.3 fold; p = 0.025) increased as well. Right atrial and plasma miR-133a expression correlated positively before CPB (r = 0.288, p = 0.045) but miR-499 expression inversely (r = −0.484, p = 0.0004). There was a strong association between plasma miR-133a and miR-499 concentrations and postoperative troponin I concentrations, the marker for myocardial damage. Increased myocardial miR-133a and miR-423-5p expression together with unchanged miR-1 and miR-499 expression might suggest active release of these miRNAs rather than their origin from damaged cells.

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“…Levels of selected microRNA were evaluated by qPCR. To remove heparin traces, RNA was treated with heparinase (Sigma) as was described before ( 41 ). For reverse transcription and qPCR microRNA-specific TaqMan Assays, TaqMan MicroRNA Reverse Transcription Kit and TaqMan Universal Master Mix II no UNG (all Thermo Fisher Scientific) were used according to manufacturer's recommendations.…”

Section: Methodsmentioning

confidence: 99%

Different Expressions of Pericardial Fluid MicroRNAs in Patients With Arrhythmogenic Right Ventricular Cardiomyopathy and Ischemic Heart Disease Undergoing Ventricular Tachycardia Ablation

Khudiakov

Panshin

Fomicheva

et al. 2021

Front. Cardiovasc. Med.

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Introduction: Pericardial fluid is enriched with biologically active molecules of cardiovascular origin including microRNAs. Investigation of the disease-specific extracellular microRNAs could shed light on the molecular processes underlying disease development. Arrhythmogenic right ventricular cardiomyopathy (ARVC) is an inherited heart disease characterized by life-threatening arrhythmias and progressive heart failure development. The current data about the association between microRNAs and ARVC development are limited.Methods and Results: We performed small RNA sequence analysis of microRNAs of pericardial fluid samples obtained during transcutaneous epicardial access for ventricular tachycardia (VT) ablation of six patients with definite ARVC and three post-infarction VT patients. Disease-associated microRNAs of pericardial fluid were identified. Five microRNAs (hsa-miR-1-3p, hsa-miR-21-5p, hsa-miR-122-5p, hsa-miR-206, and hsa-miR-3679-5p) were found to be differentially expressed between patients with ARVC and patients with post-infarction VT. Enrichment analysis of differentially expressed microRNAs revealed their close linkage to cardiac diseases.Conclusion: Our data extend the knowledge of pericardial fluid microRNA composition and highlight five pericardial fluid microRNAs potentially linked to ARVC pathogenesis. Further studies are required to confirm the use of pericardial fluid RNA sequencing in differential diagnosis of ARVC.

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Heparinase treatment of heparin-contaminated plasma from coronary artery bypass grafting patients enables reliable quantification of microRNAs

Abstract: Graphical abstract

Cited by 30 publications

References 19 publications

Mapping of genomic EGFRvIII deletions in glioblastoma: insight into rearrangement mechanisms and biomarker development

Mapping of genomic EGFRvIII deletions in glioblastoma: insight into rearrangement mechanisms and biomarker development

Establishment of an easy and straight forward heparinase protocol to analyse circulating and myocardial tissue micro-RNA during coronary artery-bypass-graft surgery

Different Expressions of Pericardial Fluid MicroRNAs in Patients With Arrhythmogenic Right Ventricular Cardiomyopathy and Ischemic Heart Disease Undergoing Ventricular Tachycardia Ablation

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