Expression Levels Of Circulating miRNAs Research Articles

Predictive value of miR-7110-5p and miR-223-3p as biomarkers for sepsis secondary to pneumonia.

Investigating the secondary sepsis of pneumonia is of great significance for rapid diagnosis and early treatment of sepsis. This study aimed to investigate the predictive value of micro ribonucleic acids (miRNA) 7110-5p and miR-223-3p in sepsis secondary to pneumonia. A miRNA microarray was used to analyze the differences in miRNA expression between patients with pneumonia and those with sepsis secondary to pneumonia. The study included a total of 50 patients with pneumonia and 42 patients with sepsis secondary to pneumonia. Quantitative polymerase chain reaction analysis was conducted to measure the circulating miRNA expression levels in patients and assess their correlations with clinical characteristics and prognosis. In this study, nine miRNAs - hsa-miR-4689-5p, hsa-miR-4621-5p, hsa-miR-6740-5p, hsa-miR-7110-5p, hsa-miR-765, hsa-miR-940, hsa-miR-213-5p, hsa-miR-223-3p, and hsa-miR-122 - met the screening criteria of having a fold change ⩾ 2 or < 0.5; p< 0.01 indicated significant differences in the results. The expression levels of miR-7110-5p and miR-223-3p differed between the two patient groups, being up-regulated in the plasma of patients with sepsis secondary to pneumonia. miR-7110-5p and miR-223-3p showed higher expression levels in both patients with pneumonia and sepsis compared to healthy controls. Moreover, the receiver operating characteristic curve revealed that the areas under the curve for predicting pneumonia using miR-7110-5p were 0.781 while those for predicting sepsis secondary to pneumonia were 0.862. For miR-223-3p, the corresponding values for predicting pneumonia and sepsis secondary to pneumonia were 0.879 and 0.924, respectively. However, there were no significant differences in the levels of miR-7110-5p and miR-223-3p between the plasma of survived and deceased patients with sepsis. MiR-7110-5p and miR-223-3p have the potential to serve as biological indicators for predicting sepsis secondary to pneumonia.

Abstract P017: Early detection of ovarian cancer in high-risk individuals using circulating miRNAs: The MiDe study

Abstract Background: Epithelial ovarian cancers generally carry a poor prognosis due to late-stage presentation and a high rate of disease recurrence. In contrast, diagnosis of early-stage disease is associated with a high rate of durable remission. Prior studies using archival samples have suggested that a neural network analysis of circulating miRNA expression levels may be an informative biomarker for early-stage ovarian cancer diagnosis. Methods: The miRNA Detection Study (MiDe Study) is a prospective, observational clinical trial open to individuals in all 50 U.S. states. Persons with at least one ovary classified as being at high-risk for ovarian cancer, either due to known genetic predisposition or family history suggestive of possible hereditary breast and ovarian cancer syndrome or Lynch syndrome, are eligible to enroll. Study subjects will submit a blood sample every 6 months for up to 5 years. Neural network analysis of serum miRNA profiles will be correlated with the subsequent diagnosis of ovarian cancer or serous tubal intraepithelial carcinoma, either clinically or at the time of risk-reducing salpingectomy or salpingo-oophorectomy. The primary outcomes are the sensitivity and specificity of the neural network for anticipating a diagnosis of epithelial ovarian cancer or a cancer precursor within 2 years of the index blood sample. The goal recruitment is 1000 study subjects. Based on a presumed cancer incidence of 0.5-1% per year among this population, approximately 25 incident cancers or precursors are expected to be diagnosed over the timespan of the study. Results: Enrollment began in September 2019. As of August 1, 2022, 372 subjects had consented to the study. Among study subjects, 270/372 (72.5%) carry a known mutation in BRCA1/2, 63/372 (16.9%) carry a known mutation in a different high-risk gene (e.g., PALB2, BRIP1), and 39/372 (10.5%) are considered high-risk due to a mutation in a gene potentially related to ovarian cancer (e.g., ATM, CHEK2) or based on family history alone. To date, 215 (57.8%) participants have contributed at least one sample, among whom 90/215 (41.9%) of subjects have contributed multiple blood samples. Among the study subjects who have given a sample, 2/215 (0.9%) have been later diagnosed with ovarian cancer. Conclusions: The MiDe Study seeks to establish the feasibility of a circulating miRNA-based test for early diagnosis of epithelial ovarian cancer among high-risk individuals. The results of this study will clarify the performance of the neural network test among a high-risk population and inform subsequent interventional trials. Citation Format: Kevin M. Elias, Dipanjan Chowdhury, Judy Garber, Ryan C. Buehler. Early detection of ovarian cancer in high-risk individuals using circulating miRNAs: The MiDe study. [abstract]. In: Proceedings of the AACR Special Conference: Precision Prevention, Early Detection, and Interception of Cancer; 2022 Nov 17-19; Austin, TX. Philadelphia (PA): AACR; Can Prev Res 2023;16(1 Suppl): Abstract nr P017.

MicroRNA-365a/b-3p as a Potential Biomarker for Hypertrophic Scars.

The clinical aspects of hypertrophic scarring vary according to personal constitution and body part. However, the mechanism of hypertrophic scar (HS) formation remains unclear. MicroRNAs (miRNAs) are known to contribute to HS formation, however, their detailed role remains unknown. In this study, candidate miRNAs were identified and analyzed as biomarkers of hypertrophic scarring for future clinical applications. HSfibroblasts and normal skin fibroblasts from patients were used for profiling and validation of miRNAs. An HS mouse model with xenografted human skin on nude mice was established. The miRNA expression between normal human, normal mouse, and mouse HS skin tissues was compared. Circulating miRNA expression levels in the serum of normal mice and mice with HSs were also analyzed. Ten upregulated and twenty-one downregulated miRNAs were detected. Among these, miR-365a/b-3p and miR-16-5p were identified as candidate miRNAs with statistically significant differences; miR-365a/b-3p was significantly upregulated (p = 0.0244). In mouse studies, miR-365a/b-3p expression levels in skin tissue and serum were higher in mice with HSs than in the control group. These results indicate that miRNAs contribute to hypertrophic scarring and that miR-365a/b-3p may be considered a potential biomarker for HS formation.

Negative pressure wound therapy affects circulating plasma microRNAs in patients with diabetic foot ulceration

AimsNegative pressure wound therapy (NPWT) is commonly used in diabetic foot ulceration (DFU). The molecular mechanisms of NPWT action, particularly outside of the wound site, have not been described. We assessed NPWT’s effect on circulating miRNA expression levels in type 2 diabetes (T2DM) patients with DFU. MethodsWe examined 34 T2DM patients treated with either NPWT (n = 24) or standard therapy (ST, n = 10). The group assignment was based on clinical criteria and local practice. Next-generation sequencing-based microRNA expression was determined on the patient’s plasma collected before therapy and after 8 days. ResultsNPWT patients were similar to the ST group in terms of age, BMI, and HbA1c level; however, they differed by mean wound area (12.6 cm2 vs. 1.1 cm2 p = 0.0005). First, we analyzed the change of miRNA after NPWT or ST and observed an upregulation of let-7f-2 only in the NPWT group. Then, we analyzed the differential expression between NPWT and ST groups, looking at possible wound size effects. We found 12 differentially expressed miRNAs in pre-treatment comparison, including let-7f-2, while in post-treatment analysis we identified 28 miRNAs. The pathway enrichment analysis suggests that identified miRNAs may be involved in wound healing, particularly through angiogenesis. ConclusionWe found initial evidence that NPWT in T2DM patients with DFU affects miRNA expression in plasma. Additionally, some differences in plasma miRNA expression may be related to wound size.

Expression levels of candidate circulating microRNAs in pediatric tuberculosis

ABSTRACT Tuberculosis (TB) is a preventable and curable disease, but increased mortality and morbidity associated with TB is one of the leading causes of deaths worldwide. MicroRNAs (miRNAs) are small, non-coding RNAs known to regulate the host immune response against TB. We investigated the expression profile of candidate circulating miRNAs, which could be used as a blood biomarker for the effective diagnosis of pediatric tuberculosis. A cross-sectional comparative study was conducted, including 30 children with active-TB and 30 healthy controls (HC) in a tertiary care hospital in Puducherry. We used the SYBR green-based miScript qRT-PCR assay to analyze the expression levels of miRNAs in plasma. Further, we used the receiver operating characteristic curve (ROC) to evaluate the diagnostic value of miRNAs. Active-TB included 25 (83.3%) pulmonary TB and 5 (16.7%) extrapulmonary TB cases. We found a significant upregulation of miR-21, miR-29a, miR-31, miR-155, and downregulation of miR-146a in children with active-TB compared to HC. The ROC analysis showed an excellent diagnostic value of miRNAs as follows: miR‑31> miR‑155> miR‑146a with AUC of (95% CI) miRNAs 0.978, 0.953, and 0.903, respectively. Altered circulating miRNA expression levels could be involved in the dysregulation of the host immune response to TB. The ROC analysis indicated that miRNAs miR-31, miR-155 and miR-146a could be effective diagnostic biomarkers for the detection of active-TB in children.

Abstract 1793: Optimal quantification of a circulating miRNA profile predictive for treatment response with droplet digital PCR in patients with multi-organ metastatic colorectal cancer

Abstract Introduction: Presence of microRNAs (miRNAs) in blood specimens hold promise for a predictive biomarker for treatment response in clinical practice as they are relatively resistant to degradation. Previously, we identified a six miRNA expression signature from tumor tissue that predicted response to first line systemic treatment in patients with metastatic colorectal cancer (mCRC) when combined with clinicopathological factors. The six miRNA expression signature consists of miR-17-5p, miR-20a-5p, miR-30a-5p, miR-92a-3p, miR-92b-3p and miR-98-5p. The aim of this study is to determine whether circulating levels of these miRNAs will be predictive for treatment outcome as well. Therefore we first studied whether these six miRNAs can be adequately measured in blood samples of patients with mCRC undergoing first line chemotherapy. Droplet digital PCR (ddPCR) technology was used for optimal quantification of miRNAs compared to normal quantitative PCR, because of low abundance of tumor specific miRNAs in blood. Methods: Serum samples of 120 patients with multi-organ mCRC who participated in a multi-center randomized controlled clinical trial were analyzed. RNA was isolated with miRNeasy Serum/Plasma Advanced Kit of Qiagen. Expression levels of the six miRNA expression signature was determined with ddPCR by using the miRCURY LNA miRNA PCR system combined with EvaGreen. Primer concentration and annealing temperature were first optimized per specific miRNA assay for ddPCR. RNA spike cel-miR-39-3p was used for quality control. Analysis was performed on samples with at least a minimal amount of 10.000 obtained droplets. Absolute quantification per patient is displayed as copies per µl (cop/µl) as defined by Quantasoft software. Results: In 120 blood samples miRNAs were measured by ddPCR. The most optimal primer concentrations for the different miRNAs was 0,5 µl for cel-miR-39-3p and 1,0 µl for the other 6 miRNAs. For cel-miR-39-3p, miR-17-5p, miR-20a-5p and miR-92a-3p the most efficient annealing temperature was 53 °C and for miR-30a-5p, miR-92b-3p and miR-98-5p this was 58 °C. The mean number of obtained droplets was 15.044 (SD 1922). The highest amount of detected copies were observed in miR-17-5p (644,4 cop/µl), miR-20a-5p (1037,1 cop/µl) and miR-92a-3p (1136,0 cop/µl). Lower expression levels were observed in miR-30a-5p (9,88 cop/µl), miR-92b-3p (1,6 cop/µl) and miR-98-5p (6,9 cop/µl). Conclusion: We demonstrated that with ddPCR absolute quantification of these six miRNAs in serum samples was efficient and reliable. Further analysis will reveal whether circulating miRNA expression levels can be used as a liquid biomarker for response to first line systemic treatment in patients with multi-organ mCRC. Citation Format: Lotte Bakkerus, Dennis Poel, Elske C. Gootjes, Cornelis Verhoef, Tineke E. Buffart, Hendrik M. Verheul. Optimal quantification of a circulating miRNA profile predictive for treatment response with droplet digital PCR in patients with multi-organ metastatic colorectal cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 1793.

Abstract 4285: Intra-individual variation in circulating miRNA expression levels in human plasma samples

Abstract Circulating miRNAs as possible non-invasive biomarkers for disease risk assessment and cancer early diagnosis has attracted increasing interest. Little information, however, is available regarding the intra-individual variation of circulating miRNA levels. We measured miRNA expression in repeated plasma samples that were collected 6 to 12 months apart from 53 healthy individuals participating in a sub-study nested in the Shanghai Women's and Shanghai Men's Health Studies. Total RNA, including miRNA, was isolated from plasma using Qiagen's miRNeasy Serum/Plasma Kit. Three synthetic spike-in RNA oligos (osa-miR414, cel-miR248, and ath-miR159a) were added to control for variances in the starting material and the efficiency of RNA extraction. NanoString's Human v2 miRNA Expression Assay, which includes CodeSets of probes specific for 800 common human miRNAs, was used for the miRNA assays. The derived miRNA counts were first corrected by subtracting the background counts and then were normalized by using the average count from three spike-in oligos and the top fifty miRNAs that gave most significant hybridization signals. The intra-individual variation was evaluated by an intra-class correlation coefficient (ICC). A total of 185 miRNAs were detected in at least 10% of samples. The detectable rate varied for each miRNA, with 69 and 44 miRNAs being detected in 50% and 75% of samples, respectively. The median ICC was 0.46 for all of the 185 detectable miRNAs. Among them, 41% (75 miRNAs) had an ICC ≥0.5, and 42 miRNAs had an ICC ≥0.6. The ICC is higher for miRNAs with higher expression levels and higher detection rates, when compared to their counterparts with lower expression or detection rates. In summary, we found that 185 miRNAs were expressed in at least 10% of plasma samples, and 69 miRNAs in 50% of samples from healthy Chinese men and women, and that 75 of these miRNAs had an ICC ≥0.5. These results suggest that common circulating miRNAs can serve as reliable biomarkers for epidemiological and clinical research. Citation Format: Jie Wu, Hui Cai, Yong-Bing Xiang, Charles Matthews, Fei Ye, Wei Zheng, Qiuyin Cai, Xiao-Ou Shu. Intra-individual variation in circulating miRNA expression levels in human plasma samples. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 4285.

Atherosclerosis-Related Circulating MicroRNAs as a Predictor of Stroke Recurrence

MicroRNAs (miRNAs) are short sequenced non-coding RNAs that posttranscriptionally regulate gene expression. We investigated circulating miRNA expression levels in acute stroke patients and its relationship with future vascular event. We included acute ischemic stroke patients who admitted to a university hospital between May 1, 2011, and July 31, 2012, and the patients with vascular risk factors but not incident stroke as controls. We collected 5 ml of venous blood, and circulating miRNA levels were evaluated by quantitative real-time polymerase chain reaction. Five miRNAs (miR-17, miR-21, miR-106a, miR-126, and miR-200b), which had been reported to be related to atherosclerosis, were measured. The levels of miRNAs were compared with the presence of acute stroke, vascular risk factors, stroke subtypes, and stroke recurrence after index stroke. A total of 120 patients were included in the study, with 83 acute stroke patients. The levels of miR-17 were significantly increased in acute stroke patients, and the levels of miR-126 had positive correlation with cerebral atherosclerosis (r = 0.254, p = 0.021). Among the 83 stroke patients, eight experienced stroke recurrence during follow-up and higher level of miR-17 was associated with shorter event-free survival (p = 0.047). This study shows that the miR-17 level was elevated in acute ischemic stroke and associated with future stroke recurrence.

Plasma miRNA levels correlate with sensitivity to bone mineral density in postmenopausal osteoporosis patients

In our study, we detect the levels of three micro-RNAs (miRNAs; miR-21, miR-133a and miR-146a) in the plasma of 120 Chinese postmenopausal women who were divided into three groups (normal, osteopenia and osteoporosis) according to the T-scores. Downregulation of miR-21, as well as upregulation of miR-133a, was validated in the plasma of osteoporosis and osteopenia patients versus the normal group. The difference in expression regarding the miR-146a level in plasma among the three groups was not significant (p > 0.01). The circulating miRNA expression levels and bone mineral density (BMD) were examined during a multiple correlation analysis as a dependent variable after adjusting for age, weight and height. We have demonstrated that specific miRNAs species are significantly changed in the plasma of osteoporosis and osteopenia patients and correlated with the BMD. Our study suggested a potential use of miR-21 and miR-133a as sensitive and plasma biomarkers for postmenopausal osteoporosis.

Causes and Consequences of MicroRNA Dysregulation in Neurodegenerative Diseases.

Neurodegenerative diseases, including Alzheimer's disease (AD), Parkinson's disease (PD), Huntington's disease (HD) and amyotrophic lateral sclerosis (ALS), originate from a loss of neurons in the central nervous system (CNS) and are severely debilitating. The incidence of neurodegenerative diseases increases with age, and they are expected to become more common due to extended life expectancy. Because of no clear mechanisms, these diseases have become a major challenge in neurobiology. It is well recognized that these disorders become the culmination of many different genetic and environmental influences. Prior studies have shown that microRNAs (miRNAs) are pathologically altered during the inexorable course of some neurodegenerative diseases, suggesting that miRNAs may be the contributing factor in neurodegeneration. Here, we review what is known about the involvement of miRNAs in the pathogenesis of neurodegenerative diseases. The biogenesis of miRNAs and various functions of miRNAs that act as the chief regulators will be discussed. We focus in particular on dysregulation of miRNAs which leads to several neurodegenerative diseases from three aspects: miRNA-generating disorders, miRNA-targeting genes and epigenetic alterations. Furthermore, recent evidences have shown that circulating miRNA expression levels are changed in patients with neurodegenerative diseases. Circulating miRNA expression levels are reported in patients in order to evaluate their application as biomarkers of these diseases. A discussion is included with a potential diagnostic biomarker and the possible future direction in exploring the nexus between miRNAs and various neurodegenerative diseases.

Circulating microRNA Profile throughout the Menstrual Cycle

Normal physiological variables, such as age and gender, contribute to alterations in circulating microRNA (miRNA) expression levels. The changes in the female body during the menstrual cycle can also be reflected in plasma miRNA expression levels. Therefore, this study aimed to determine the plasma miRNA profile of healthy women during the menstrual cycle and to assess which circulating miRNAs are derived from blood cells. The plasma miRNA expression profiles in nine healthy women were determined by quantitative real time PCR using Exiqon Human Panel I assays from four time-points of the menstrual cycle. This platform was also used for studying miRNAs from pooled whole blood RNA samples at the same four time-points. Our results indicated that circulating miRNA expression levels in healthy women were not significantly altered by the processes occurring during the menstrual cycle. No significant differences in plasma miRNA expression levels were observed between the menstrual cycle time-points, but the number of detected miRNAs showed considerable variation among the studied individuals. miRNA analysis from whole blood samples revealed that majority of miRNAs in plasma are derived from blood cells. The most abundant miRNA in plasma and blood was hsa-miR-451a, but a number of miRNAs were only detected in one or the other sample type. In conclusion, our data suggest that the changes in the female body during the menstrual cycle do not affect the expression of circulating miRNAs at measurable levels.