103 CIRCULATING microRNAs FOR EARLY DIAGNOSIS OF BOVINE PREGNANCY

2015 ◽  
Vol 27 (1) ◽  
pp. 144
Author(s):  
J. Ioannidis ◽  
C. Ashworth ◽  
R. Raue ◽  
X. Donadeu
Keyword(s):  
Early Diagnosis ◽  
Deep Sequencing ◽  
Mirna Expression ◽  
Plasma Samples ◽  
Circulating Micrornas ◽  
Circulating Mirna ◽  
Time Points ◽  
Bovine Plasma ◽  
Mirna Sequencing ◽  
Pooled Samples

Early diagnosis of pregnancy can shorten calving intervals, improve annual milk production and increase overall profits from modern dairy herds. At present, accurate diagnosis is only possible after the third week of pregnancy. Circulating microRNAs (miRNAs) have been proposed as diagnostic biomarkers for numerous human conditions such as cancer and diabetes. Moreover, distinct circulating miRNA profiles have been associated with different stages of human pregnancy. The objective of this study was to determine whether differential miRNA profiles occur in circulation during early pregnancy (Day 24 or earlier) in cattle that could be used for diagnostic purposes. Holstein-cross heifers were oestrous-synchronised and artificially inseminated (AI, n = 11) or sham-inseminated (control, n = 8) at first detected oestrus. Plasma samples were collected on Days 0, 8, 16 and 24 after insemination. Circulating miRNA levels were independently determined in pooled plasma samples (n = 3 pools for each of pregnant Day 24 and nonpregnant Days 0, 8, and 16) using Qiagen qPCR arrays (Qiagen, Valencia, CA, USA) and in individual samples (n = 11 samples for each pregnant Days 16 and 24, and 8 samples for each of nonpregnant Days 0, 8, and 16, respectively) using Illumina miRNA sequencing. The qPCR array data were analysed using the ΔΔCq method. The miRNA sequencing data were normalised using EdgeR. Differential expression between pregnant and nonpregnant groups was determined using 2-sample t-tests with false discovery rate (FDR) adjustment. Differences in miRNA expression were validated by RT-qPCR. Out of a total of 191 miRNAs analysed in pooled samples using qPCR arrays, 8 were differentially expressed (<3-fold, FDR <0.1) in Day 24 pregnant heifers relative to nonpregnant heifers (Days 0, 8, and 16 combined). No miRNAs were differentially expressed (FDR >0.1) between nonpregnant time-points. Changes in levels of 11 miRNAs were validated by RT-qPCR in individual plasma samples; although expression trends for these miRNAs were the same as in pooled samples, none of the changes in individual samples were significant after FDR adjustment (P > 0.1). Deep sequencing (96 million miRNA reads) identified 231 miRNAs in bovine plasma. There were no significant differences (FDR >0.1) in the expression of any miRNAs between pregnant heifers (Days 16 or 24) and nonpregnant (Days 0, 8, and 16 individually or combined). In addition, no significant differences were identified among nonpregnant time-points. In summary, we successfully performed miRNA profiling of bovine plasma using both deep sequencing and qPCR; however, we did not detect differences in miRNA expression between early pregnant (Day 16 or 24) and nonpregnant heifers. Changes in circulating miRNA levels may involve low abundance miRNAs that cannot be accurately quantified using current technology. Alternatively, changes in circulating miRNA levels may only occur later during pregnancy in cattle.

scholarly journals Circulating microRNAs correlate to clinical parameters in individuals with allergic and non-allergic asthma

2020 ◽  
Vol 21 (1) ◽  
Author(s):  
Julie Weidner ◽  
Linda Ekerljung ◽  
Carina Malmhäll ◽  
Nicolae Miron ◽  
Madeleine Rådinger
Keyword(s):  
Lung Function ◽  
Personalized Medicine ◽  
Allergic Asthma ◽  
Mirna Expression ◽  
Mirna Gene ◽  
Blood Eosinophil ◽  
Clinical Parameters ◽  
Inhaled Corticosteroid ◽  
Circulating Micrornas ◽  
Circulating Mirna

Abstract Background Asthma is a chronic airway disease affecting millions of people. Better methods to define asthma subgroups using clinical parameters and molecular biomarkers are crucial in the development of personalized medicine. Objective The aim of this study was to determine if circulating microRNAs (miRNAs) may be used to distinguish well–defined asthma groups. Methods Blood serum from 116 well-defined subjects, including healthy controls and individuals with allergic or non-allergic asthma, from the West Sweden Asthma Study were included. Serum was analyzed for circulating miRNA expression of miR-126, − 145, −146a, − 155, − 223, and -374a and eosinophil cationic protein (ECP). Correlations between clinical characteristics and circulating miRNA expression as well as potential miRNA gene targets were investigated. Results A subset of miRNAs were differentially expressed between allergic and non-allergic asthmatic individuals. Alterations in expression of miR-155, −146a, −374a and − 145 were observed in allergic asthmatics in response to inhaled corticosteroid usage. Additionally, miR-223 and miR-374a expression varied in non-allergic asthmatics based on blood eosinophil numbers. Numerous clinical parameters, including lung function measurements, correlated with subsets of miRNAs. Finally, pathway analysis revealed a potential role for inhaled corticosteroid induced miRNAs in leukocyte regulation, IL-6 signaling and glucocorticoid response. Conclusion Circulating miRNA expression was altered in subjects with allergic and non-allergic asthma and correlated to clinical parameters including lung function and potential gene targets involved in immune processes. This combination of clinical and molecular data may be a basis for the further, more precise classification of asthma subgroups. Taken together, these findings would further asthma research and benefit future patients through the discovery of molecular mechanisms as well as identifying asthma subgroups contributing to the development of personalized medicine.

scholarly journals PlasmiR: A Manual Collection of Circulating microRNAs of Prognostic and Diagnostic Value

Cancers ◽  
2021 ◽  
Vol 13 (15) ◽  
pp. 3680
Author(s):  
Spyros Tastsoglou ◽  
Marios Miliotis ◽  
Ioannis Kavakiotis ◽  
Athanasios Alexiou ◽  
Eleni C. Gkotsi ◽  
...  
Keyword(s):  
Circulatory System ◽  
Transcriptional Regulators ◽  
Diagnostic Value ◽  
Plasma Samples ◽  
Online Database ◽  
Exclusion Criteria ◽  
Circulating Micrornas ◽  
Circulating Mirna ◽  
Meta Information ◽  
Non Coding Rnas

Only recently, microRNAs (miRNAs) were found to exist in traceable and distinctive amounts in the human circulatory system, bringing forth the intriguing possibility of using them as minimally invasive biomarkers. miRNAs are short non-coding RNAs that act as potent post-transcriptional regulators of gene expression. Extensive studies in cancer and other disease landscapes investigate the protective/pathogenic functions of dysregulated miRNAs, as well as their biomarker potential. A specialized resource amassing experimentally verified, circulating miRNA biomarkers does not exist. We queried the existing literature to identify articles assessing diagnostic/prognostic roles of miRNAs in blood, serum, or plasma samples. Articles were scrutinized in order to exclude instances lacking sufficient experimental documentation or employing no biomarker assessment methods. We incorporated information from more than 200 biomedical articles, annotating crucial meta-information including cohort sizes, inclusion-exclusion criteria, disease/healthy confirmation methods and quantification details. miRNAs and diseases were systematically characterized using reference resources. Our circulating miRNA biomarker collection is provided as an online database, plasmiR. It consists of 1021 entries regarding 251 miRNAs and 112 diseases. More than half of plasmiR’s entries refer to cancerous and neoplastic conditions, 183 of them (32%) describing prognostic associations. plasmiR facilitates smart queries, emphasizing visualization and exploratory modes for all researchers.

scholarly journals ATIM-43. PLASMA EXTRACELLULAR VESICLE MIRNA SIGNATURES IN GBM PATIENTS RECEIVING AN EXPERIMENTAL IMMUNOTHERAPY

2019 ◽  
Vol 21 (Supplement_6) ◽  
pp. vi11-vi11
Author(s):  
Luz Cumba-Garcia ◽  
Mrunal Dehankar ◽  
Asha Nair ◽  
Ian Parney
Keyword(s):  
Clinical Trial ◽  
Treatment Response ◽  
Mirna Expression ◽  
Malignant Gliomas ◽  
Extracellular Vesicle ◽  
Mirna Expression Profile ◽  
P Value ◽  
Time Points ◽  
Mirna Sequencing ◽  
Monitor Treatment Response

Abstract Patients with glioblastoma (GBM) have a median survival of 15 months despite aggressive treatment. Immunotherapies such as dendritic cell (DC) vaccines have modest clinical efficacy in small clinical trials. Treatment-related pseudo-progression confounds outcome assessment by MRI, particularly in patients receiving immunotherapy. Thus, there is a need for additional non-invasive methods to monitor treatment response. Extracellular vesicles (EVs), especially plasma exosomes, contain tumor-specific microRNA (miRNA) cargo that could serve as a liquid biopsy to distinguish true progression from treatment-related pseudo-progression. Plasma exosomes were isolated by serial density gradient ultracentrifugation from 20 newly diagnosed GBM patients enrolled in a clinical trial of allogeneic tumor lysate-pulsed autologous DC vaccination. Short non-coding RNA sequencing and bioinformatics analysis was performed for each patient at three time points (TP): pre-vaccine (TP1), post-initial vaccine (TP2), and at end of treatment (TP3). miRNA expression analysis revealed a total of 14 upregulated and 12 downregulated miRNAs across time points (p-value < 0.05, |logFC| >1), few of which have been previously reported to be differentially expressed in GBM. Interestingly, patients’ miRNA profile expression differed more at the beginning of treatment (e.g. TP1-vs-TP2) and at subsequent time points (e.g. TP2-vs-TP3). Ingenuity Pathway Analysis is in progress to identify pathways associated with immunotherapy treatment response in malignant gliomas. In conclusion, miRNA sequencing from GBM patients’ plasma exosomes enrolled in our DC clinical trial shows marked differential miRNA expression between time points. These results suggest that as patients progress through treatment, consistent differences in plasma exosomal miRNA expression profile can be identified that could be utilized as predictors of treatment response. Thus, plasma EVs may serve as a robust platform to monitor treatment outcome.

scholarly journals Integrated miRNA/cytokine/chemokine profiling reveals immunopathological step changes associated with COVID-19 severity

2021 ◽  
Author(s):  
Julie C. WIlson ◽  
David Kealy ◽  
Sally R. James ◽  
Katherine Newling ◽  
Christopher Jagger ◽  
...  
Keyword(s):  
Cell Death ◽  
Correlation Analysis ◽  
Partial Least Squares Regression ◽  
Clinical Parameters ◽  
Plasma Samples ◽  
Least Squares Regression ◽  
Circulating Micrornas ◽  
Circulating Mirna ◽  
Novel Approach ◽  
Insight Into

Circulating microRNAs (miRNAs) are exceptional mechanism-based correlates of disease, yet their potential remains largely untapped in COVID-19. Here, we determined circulating miRNA and cytokine and chemokine (CC) profiles in 171 blood plasma samples from 58 hospitalised COVID-19 patients. Thirty-two miRNAs were differentially expressed in severe cases when compared to moderate and mild cases. These miRNAs and their predicted targets reflected key COVID-19 features including cell death and hypoxia. Compared to mild cases, moderate and severe cases were characterised by a global decrease in circulating miRNA levels. Partial least squares regression using miRNA and CC measurements allowed for discrimination of severe cases with greater accuracy (87%) than using miRNA or CC levels alone. Correlation analysis revealed severity group-specific associations between CC and miRNA levels. Importantly, the miRNAs that correlated with IL6 and CXCL10, two cardinal COVID-19-associated cytokines, were distinct between severity groups, providing a novel qualitative way to stratify patients with similar levels of proinflammatory cytokines but different disease severity. Integration of miRNA and CC levels with clinical parameters revealed severity-specific signatures associated with clinical hallmarks of COVID-19. Our study highlights the existence of severity-specific circulating CC/miRNA networks, providing insight into COVID-19 pathogenesis and a novel approach for monitoring COVID-19 progression.

scholarly journals Direct supplementation with Urolithin A overcomes limitations of dietary exposure and gut microbiome variability in healthy adults to achieve consistent levels across the population

2021 ◽  
Author(s):  
Anurag Singh ◽  
Davide D’Amico ◽  
Pénélope A. Andreux ◽  
Gillian Dunngalvin ◽  
Timo Kern ◽  
...  
Keyword(s):  
Gut Microbiome ◽  
Dietary Exposure ◽  
Food Product ◽  
Pomegranate Juice ◽  
Healthy Population ◽  
Plasma Samples ◽  
Time Points ◽  
Natural Exposure ◽  
Microbiome Diversity ◽  
Urolithin A

Abstract Background Urolithin A (UA) is produced by gut microflora from foods rich in ellagitannins. UA has been shown to improve mitochondrial health preclinically and in humans. Not everyone has a microbiome capable of producing UA, making supplementation with UA an appealing strategy. Objective This is the first detailed investigation of the prevalence of UA producers in a healthy population and the ability of direct UA supplementation to overcome both microbiome and dietary variability. Dietary intake of a glass of pomegranate juice (PJ) was used to assess UA producer status (n = 100 participants) and to characterize differences in gut microbiome between UA producers from non-producers. Methods Subjects were randomized (1:1) to either PJ or a food product containing UA (500 mg). Prevalence of UA producers and non-producers were determined in the PJ group. Diet questionnaires and fecal samples were collected to compare differences between UA producers and non-producers along with plasma samples at different time points to assess levels of UA and its conjugates between the interventions. Results Only 12% of subjects had detectable levels of UA at baseline. Following PJ intake ~40% of the subjects converted significantly the precursor compounds into UA. UA producers were distinguished by a significantly higher gut microbiome diversity and ratio of Firmicutes to Bacteroides. Direct supplementation with UA significantly increased plasma levels and provided a >6-fold exposure to UA vs. PJ (p < 0.0001). Conclusions Differences in gut microbiome and diet that dictate natural exposure to UA can be overcome via direct dietary UA supplementation.

Circulating miR-126 as a Potential Non-invasive Biomarker for Intracranial Aneurysmal Rupture: A Pilot Study

2021 ◽  
Vol 19 ◽  
Author(s):  
Jianing Wu ◽  
Ilgiz Gareev ◽  
Ozal Beylerli ◽  
Albert Mukhamedzyanov ◽  
Valentin Pavlov ◽  
...  
Keyword(s):  
Aneurysmal Subarachnoid Hemorrhage ◽  
Diagnostic Value ◽  
Plasma Samples ◽  
Expression Levels ◽  
Time Points ◽  
Non Invasive ◽  
Risk Of Rupture ◽  
Life Threatening ◽  
Healthy Control ◽  
New Biomarkers

Aim: Intracranial aneurysms (IAs) are characterized by abnormal dilation and thinning of the cerebral vessels wall, leading to rupture and life-threatening aneurysmal subarachnoid hemorrhage (aSAH) condition. This dictates the need to find new biomarkers that predict the presence of IAs and the risk of their rupture. The aim of this study was to measure circulating miR-126 at various time points post-aSAH to identify the timing of peak levels. Methods: Plasma samples from 62 patients with unruptured IAs (UIAs), 80 patients with aSAH at various time points (1, 3, 7, and 14 days post-event), and 47 healthy control were collected and subjected to qRT-PCR analyses for the expression levels of circulating miR-126. ROC curve and AUC were used to evaluate the diagnostic value of circulating miR-126. Results: The expression levels of circulating miR-126 were increased in patients with UIAs than in the healthy control. Furthermore, the expression levels of circulating miR-126 rose substantially from day 1 to day 7, but with a moderate decrease from day 7 to day 14 in plasma of patients with aSAH. The peak was observed on day 7. The AUC for miR-126 was 0.75, 0.75, 0.82, 0.87, and 0.79, respectively, and demonstrated that circulating miR-126 displayed considerable accuracy in discriminating plasma of patients with UIAs and patients after aSAH at various time points from a healthy control. Conclusion: Our results indicated that circulating miR-126 in plasma samples could be served as a potential non-invasive biomarker in IAs detection and prevention IAs with a high risk of rupture.

scholarly journals Detection of Neprilysin-Derived BNP Fragments in the Circulation: Possible Insights for Targeted Neprilysin Inhibition Therapy for Heart Failure

2019 ◽  
Vol 65 (10) ◽  
pp. 1239-1247 ◽  
Author(s):  
Evgeniya E Feygina ◽  
Marina M Artemieva ◽  
Alexander B Postnikov ◽  
Natalia N Tamm ◽  
Marina N Bloshchitsyna ◽  
...  
Keyword(s):  
Heart Failure ◽  
Ring Opening ◽  
Active Form ◽  
Cross Reactivity ◽  
Plasma Samples ◽  
Time Points ◽  
Positive Effects ◽  
Edta Plasma ◽  
Neprilysin Inhibition ◽  
Nep Inhibition

Abstract BACKGROUND Entresto™ is a new heart failure (HF) therapy that includes the neprilysin (NEP) inhibitor sacubitril. One of the NEP substrates is B-type natriuretic peptide (BNP); its augmentation by NEP inhibition is considered as a possible mechanism for the positive effects of Entresto. We hypothesized that the circulating products of BNP proteolysis by NEP might reflect NEP impact on the metabolism of active BNP. We suggest that NEP-based BNP cleavage at position 17–18 results in BNP ring opening and formation of a novel epitope with C-terminal Arg-17 (BNP-neo17 form). In this study, we use a specific immunoassay to explore BNP-neo17 in a rat model and HF patient plasma. METHODS We injected BNP into rats, with or without NEP inhibition with sacubitril. BNP-neo17 in plasma samples at different time points was measured with a specific immunoassay with neglectable cross-reactivity to intact forms. BNP-neo17 and total BNP were measured in EDTA plasma samples of HF patients. RESULTS BNP-neo17 generation in rat circulation was prevented by NEP inhibition. The maximum 13.2-fold difference in BNP-neo17 concentrations with and without sacubitril was observed at 2 min after injection. BNP-neo17 concentrations in 32 HF patient EDTA plasma samples ranged from 0 to 37 pg/mL (median, 5.4; interquartile range, 0–9.1). BNP-neo17/total BNP had no correlation with total BNP concentration (with r = −0.175, P = 0.680) and showed variability among individuals. CONCLUSIONS BNP-neo17 formation is NEP dependent. Considering that BNP-neo17 is generated from the active form of BNP by NEP, we speculate that BNP-neo17 may reflect both the NEP activity and natriuretic potential and serve for HF therapy guidance.

scholarly journals Monitoring Radiotherapeutic Response in Prostate Cancer Patients Using High Throughput FTIR Spectroscopy of Liquid Biopsies

Cancers ◽  
2019 ◽  
Vol 11 (7) ◽  
pp. 925 ◽  
Author(s):  
Dinesh K.R. Medipally ◽  
Thi Nguyet Que Nguyen ◽  
Jane Bryant ◽  
Valérie Untereiner ◽  
Ganesh D. Sockalingum ◽  
...  
Keyword(s):  
Prostate Cancer ◽  
Blood Plasma ◽  
Ftir Spectroscopy ◽  
Cancer Patients ◽  
Radiation Toxicity ◽  
Plasma Samples ◽  
Analysis Model ◽  
Liquid Biopsies ◽  
Time Points ◽  
Wide Range

Radiation therapy (RT) is used to treat approximately 50% of all cancer patients. However, RT causes a wide range of adverse late effects that can affect a patient’s quality of life. There are currently no predictive assays in clinical use to identify patients at risk of normal tissue radiation toxicity. This study aimed to investigate the potential of Fourier transform infrared (FTIR) spectroscopy for monitoring radiotherapeutic response. Blood plasma was acquired from 53 prostate cancer patients at five different time points: prior to treatment, after hormone treatment, at the end of radiotherapy, two months post radiotherapy and eight months post radiotherapy. FTIR spectra were recorded from plasma samples at all time points and the data was analysed using MATLAB software. Discrimination was observed between spectra recorded at baseline versus follow up time points, as well as between spectra from patients showing minimal and severe acute and late toxicity using principal component analysis. A partial least squares discriminant analysis model achieved sensitivity and specificity rates ranging from 80% to 99%. This technology may have potential to monitor radiotherapeutic response in prostate cancer patients using non-invasive blood plasma samples and could lead to individualised patient radiotherapy.

scholarly journals MicroRNA profiling of mouse liver in response to DENV-1 infection by deep sequencing

PeerJ ◽  
2019 ◽  
Vol 7 ◽  
pp. e6697 ◽  
Author(s):  
Lian Yih Pong ◽  
Sinikka Parkkinen ◽  
Amreeta Dhanoa ◽  
Han Ming Gan ◽  
Indeevari Abisheka Chiharu Wickremesinghe ◽  
...  
Keyword(s):  
Immune Responses ◽  
Infected Mouse ◽  
Deep Sequencing ◽  
Molecular Mechanisms ◽  
Fold Change ◽  
Differentially Expressed ◽  
Pathway Enrichment Analysis ◽  
Adaptive Immune Responses ◽  
Adaptive Immune ◽  
Mirna Sequencing

BackgroundDengue caused by dengue virus (DENV) serotypes −1 to −4 is the most important mosquito-borne viral disease in the tropical and sub-tropical countries worldwide. Yet many of the pathophysiological mechanisms of host responses during DENV infection remain largely unknown and incompletely understood.MethodsUsing a mouse model, the miRNA expressions in liver during DENV-1 infection was investigated using high throughput miRNA sequencing. The differential expressions of miRNAs were then validated by qPCR, followed by target genes prediction. The identified miRNA targets were subjected to gene ontology (GO) annotation and pathway enrichment analysis to elucidate the potential biological pathways and molecular mechanisms associated with DENV-1 infection.ResultsA total of 224 and 372 miRNAs out of 433 known mouse miRNAs were detected in the livers of DENV-1-infected and uninfected mice, respectively; of these, 207 miRNAs were present in both libraries. The miR-148a-3p and miR-122-5p were the two most abundant miRNAs in both groups. Thirty-one miRNAs were found to have at least 2-fold change in upregulation or downregulation, in which seven miRNAs were upregulated and 24 miRNAs were downregulated in the DENV-1-infected mouse livers. The miR-1a-3p was found to be the most downregulated miRNA in the DENV-1-infected mouse livers, with a significant fold change of 0.10. To validate the miRNA sequencing result, the expression pattern of 12 miRNAs, which were highly differentially expressed or most abundant, were assessed by qPCR and nine of them correlated positively with the one observed in deep sequencing.In silicofunctional analysis revealed that the adaptive immune responses involving TGF-beta, MAPK, PI3K-Akt, Rap1, Wnt and Ras signalling pathways were modulated collectively by 23 highly differentially expressed miRNAs during DENV-1 infection.ConclusionThis study provides the first insight into the global miRNA expressions of mouse livers in response to DENV-1 infectionin vivoand the possible roles of miRNAs in modulating the adaptive immune responses during DENV-1 infection.

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