Identification of circular RNA hsa_circ_0034621 as a novel biomarker for carotid atherosclerosis and the potential function as a regulator of NLRP3 inflammasome.

BACKGROUND AND AIMS: NLRP3 inflammasome plays a key role in vascular inflammation and atherosclerosis. Circular RNAs (circRNAs) are involved in disease development by regulating gene expression, and have emerged as promising novel disease biomarkers. This study aimed to identify the NLRP3 inflammasome-associated circRNA biomarkers of carotid atherosclerosis. METHODS: Based on the differential expression profiles of circRNAs in patients with carotid artery plaque (CAP) and healthy controls, hsa_circ_0043621, hsa_circ_0051995, and hsa_circ_0123388 were screened and validated using real-time quantitative polymerase chain reaction (RT-qPCR). Potential circRNA-miRNA-mRNA interactions were explored using a luciferase assay. The biological roles of the validated circRNAs were investigated in human umbilical vein endothelial cells (HUVECs) using Western blotting, transwell, and CCK-8 assays. Clinical significance was assessed using receiver operating characteristic (ROC) curves and logistic regression analysis. RESULTS: The expression levels of all candidate circRNAs were significantly higher in patients with CAP than in controls (p<0.05), which was consistent with the results of the microarray analysis. Overexpression of hsa_circ_0043621 significantly increased the expression of NLRP3, induced migration of HUVECs, and inhibited cell proliferation. hsa_circ_0043621 demonstrated reasonable diagnostic accuracy for CAP detection and increased intima-media thickness (IMT). hsa_circ_0043621 upregulation was an independent predictor of an increased risk of CAP and increased IMT. CONCLUSIONS: hsa_circ_0043621 is a valuable circulating biomarker of carotid atherosclerosis and may contribute to its pathogenesis by regulating the NLRP3 inflammasome.

Association between systemic immune inflammatory/inflammatory response index and hypertension: A cohort study of functional community.

BACKGROUND AND AIMS: In prospective studies, there is limited evidence of the association between inflammation and hypertension. We aimed to explore the relationship between systemic immune inflammatory index (SII)/systemic inflammatory response index (SIRI) and hypertension in a prospective cohort study to identify the best inflammatory cell markers that predict hypertension. METHODS AND RESULTS: This study was conducted in a functional community cohort in Beijing. In 2015, a total of 6003 individuals without hypertension were recruited and followed up until 2021. Using a restriction cubic spline with baseline SII/SIRI as a continuous variable, the dose-response relationship between hypertension and SII/SIRI was explored. Logistic regression was used to analyze the correlation between hypertension and SII/SIRI trajectory groups. At a mean follow-up of 6 years, 970 participants developed hypertension. SII showed a significant nonlinear dose-response relationship with hypertension (P < 0.05). Higher SII/SIRI was associated with an increased risk of hypertension (SII: RR = 1.003, 95%CI: 1.001-1.004; SIRI: RR = 1.228, 95%CI: 1.015-1.486). Both SII and SIRI were more predictive in males than females (SII: 0.698 vs. 0.695; SIRI: 0.686 vs. 0.678). CONCLUSION: Both systemic immune inflammatory index (SII) and systemic inflammatory response Index (SIRI) independently increased the risk of hypertension, and both were effective inflammatory cell indicators that predict the risk of hypertension.

Occupational stress is associated with insulin resistance and incident type 2 diabetes: A prospective cohort study of functional community.

BACKGROUND: To exploit the association of occupational stress with the development of insulin resistance (IR) and type 2 diabetes (T2D) in a Chinese population-based cohort. METHODS: A total of 6109 participants from a functional community cohort in Beijing were enrolled in 2015 and followed up until 2021. Copenhagen Psychosocial Questionnaire (COPSOQ) were used to evaluate occupational stress. RESULTS: At baseline, increase values of all five scales of COPSOQ and total COPSOQ were significantly associated with IR. During an average 5.63 y follow-up, 732 individuals developed T2D. Increasing in values of "Demands at work", "Insecurity at work", "Job satisfaction" and total COPSOQ were significantly associated with incident T2D (P < 0.01). Mediation analysis showed that subjectively perceived occupational stress promoted T2D mainly by affecting plasma cortisol and the mediation effects of HOMA-IR, SBP, DBP, TG, Urea and UA were significant on the association between cortisol and incident T2D, with proportion mediated of 37.1%, 8.12%, 2.02%, 2.94%, 2.35% and 2.70%. CONCLUSION: Occupational stress was independently associated with the development of IR and T2D. IR, BP, TG, Urea and UA all partly mediated the association between occupational stress and incident T2D.

CircRNA hsa_circ_0071336 is associated with type 2 diabetes through targeting the miR-93-5p/GLUT4 axis.

Glucose transporter 4 (GLUT4) is a dominant regulator of whole-body glucose homeostasis. Accumulating evidence has shown that circular RNAs (circRNAs) play significant roles in the pathogenesis of disease. The aim of the present study was to identify the circRNA that can be used as a novel biomarker for type 2 diabetes (T2D) through regulating GLUT4. Based on previous microarray analysis comparing T2D cases and healthy controls, hsa_circ_0071336, which was predicted to be a regulator of GLUT4 by acting as a competitive endogenous RNAs (ceRNA) to sponge miR-93-5p, was selected for further validation. The clinical significance of circulating hsa_circ_0071336 was investigated in a large independent cohort. The results showed that circulating hsa_circ_0071336 was significantly downregulated in blood in T2D and had a high diagnostic accuracy for discriminating T2D and impaired fasting glucose (IFG) from healthy controls. Low expression of circ_0071336 was an independent predictor of T2D, IFG and insulin resistance. A luciferase reporter assay and western-blot analysis indicated that miR-93-5p was a direct target of hsa_circ_0071336, and miR-93-5p may negatively regulate the expression of GLUT4. The expression levels of hsa_circ_007136 were negatively related to miR-93-5p expression and positively correlated with the mRNA expression of GLUT4 in adipose tissues. In conclusion, hsa_circRNA_0071336 can be considered as a potential novel and stable biomarker for T2D and its early detection. hsa_circ_0071336 regulates the GLUT4 expression by sponging miR-93-5p and maybe involved in the pathogenesis of T2D. These findings may unveil new targets for the prevention, diagnosis and treatment of T2D.

Pri-miR-144 rs9279 is associated with type 2 diabetes and regulation of stress response.

Single-nucleotide polymorphisms (SNPs) of microRNAs (miRNAs) may alter miRNA expression, binding affinity, and/or messenger RNA expression levels of the target genes, thus leading to disease susceptibility. This study explored the association between SNPs in neuroendocrine stress response-related miRNAs and type 2 diabetes (T2D). In the screening stage, the association between six candidate SNPs of miRNAs and T2D was analyzed in a case-control study including 504 T2D cases and 494 healthy controls. Homozygous GG genotype of pri-miR-144 rs9279 showed significant association with increased risk of T2D compared with homozygous TT genotype (adjusted odds ratio [OR] = 1.62, 95% confidence interval [CI]: 1.07-2.45; p = .023) and the combined TT/TG genotype (adjusted OR = 1.59, 95% CI: 1.08-2.36; p = .020). In the validation stage, the association was further validated in a second independent set of subjects. The GG genotype showed consistent directions and effect sizes that were identified in previous additive and recessive models. The expression levels of miRNAs were further compared between different genotypes in the 179 newly diagnosed cases and 183 frequency-matched healthy controls. As a result, the GG genotype carriers had significantly upregulated expression of plasma miR-144 and cortisol, as compared to individuals with TT and TG genotypes, respectively, in total subjects and subgroups (p < .05). Eventually, NR3C1 was proved to be a stress-related target gene of miR-144, indicating that pri-miR-144 rs9279 may contribute to the development of T2D by altering regulation of stress response.

hsa_circ_0111707 Is Associated With Risk of Stress-Related Type 2 Diabetes via Sponging miR-144-3p.

Introduction: Chronic stress plays an important role in the development of type 2 diabetes (T2D). Circular RNAs (circRNAs) play significant roles in regulating the pathogenesis of diseases by regulating gene expression. The aim of the present study was to identify the association between hsa_circ_0111707 and stress-related T2D. Methods: The present study was performed based on a three-part design. The association between hsa_circ_0111707 in peripheral blood mononuclear cells (PBMCs) and T2D and stress-related variables were assessed in a cross-sectional study. The causal relationship of hsa_circ_0111707 on T2D was further investigated in a nested case-control study. miR-144-3p as the miRNA target of hsa_circ_0111707 was verified by dual-luciferase reporter assay and RNA immunoprecipitation (RIP) assay. Results: The relative expression of hsa_circ_0111707 was significantly lower in the T2D and impaired fasting glucose (IFG) cases in comparison with controls. The hsa_circ_0111707 expression was significantly negatively correlated with miR-144-3p expression and plasma cortisol concentration and positively correlated with NR3C1 expression. In addition, hsa_circ_0111707 expression was negatively correlated with scores of "demands at work" and "insecurity at work" of Copenhagen Psychosocial Questionnaire (COPSOQ). Decreased hsa_circ_0111707 expression was associated with increased risk of T2D development. Functional analysis demonstrated that hsa_circ_0111707 functions as a sponge for miR-144-3p. Conclusion: hsa_circ_0111707 is associated with risk of T2D development via sponging miR-144-3p. hsa_circ_0111707 in PBMCs can be considered a potential biomarker of stress-related T2D.

Identification of stress-related microRNA biomarkers in type 2 diabetes mellitus: A systematic review and meta-analysis.

BACKGROUND: Many studies have investigated microRNAs (miRNAs) in the detection of type 2 diabetes mellitus (T2DM). Herein, the dysregulated direction of stress-related miRNAs used as biomarkers of T2DM are summarized and analyzed. METHODS: PubMed, EMBASE, ISI Web of Science, and three Chinese databases were searched for case-control miRNA profiling studies about T2DM. A meta-analysis under a random effect was performed. Subgroup analysis was conducted based on different tissues and species. Sensitivity analysis was conducted to confirm the robustness among studies. The effect size was pooled using ln odds ratios (ORs), 95% confidence intervals (95% CIs), and P-values. RESULTS: The present meta-analysis included 39 case-control studies with a total of 494 miRNAs. Only 33 miRNAs were reported in three or more studies and, of these, 18 were inconsistent in their direction of dysregulation. Two significantly dysregulated miRNAs (let-7 g and miR-155) were identified in the meta-analysis. Four miRNAs (miR-142-3p, miR-155, miR-21, and miR-34c-5p) were dysregulated in patients with T2DM, whereas five miRNAs (miR-146a, miR-199a-3p, miR-200b, miR-29b and miR-30e) were dysregulated in animal models of diabetes. In addition, two dysregulated miRNAs (miR-146a and miR-21) were highly cornea specific and heart specific. In sensitivity analysis, only miR-155 was still significantly dysregulated after removing studies with small sample sizes. CONCLUSIONS: The present meta-analysis revealed that 16 stress-related miRNAs were significantly dysregulated in T2DM. MiR-148b, miR-223, miR-130a, miR-19a, miR-26b and miR-27b were selected as potential circulating biomarkers of T2DM. In addition, miR-146a and miR-21 were identified as potential tissue biomarkers of T2DM.

A meta-analysis of dysregulated miRNAs in coronary heart disease.

AIMS: To combine the results of dysregulated miRNAs in individual coronary heart disease (CHD) studies and to identify potential miRNA biomarkers. MAIN METHODS: MiRNA profiling studies of CHD were extracted from Pubmed, Embase, Web of Science and China National Knowledge Internet (CNKI) databases if they met the inclusion criteria. The meta-analysis was conducted using a random effects model to identify the effect of each multiple-reported miRNA. We also performed subgroup analysis according to miRNA detecting methods, tissues and subtypes of CHD. Sensitivity analysis was performed on the sample size. Bioinformatic analysis was performed to identify the potential biomatic functions. All results were represented as log10 odds ratios (logORs). KEY FINDINGS: A total of 239 miRNAs were reported to be dysregulated in all 25 studies analyzed herein, and meta-analysis identified 48 statistically significant miRNAs. Bioinformatic analysis showed they were closely related with CHD. The most reported up-regulated miRNA was miR-122-5p (logOR: 2.7924, P < 0.001). A total of 7, 6, 4 and 9 miRNAs were detected to be differentially expressed in myocardial infarction (MI), unstable angia (UA), stable angina (SA) and pre-CHD subjects, respectively. 32 miRNAs were dysregulated in blood sample. The dysregulation of miR-133a-3p in whole blood and plasma/serum was contrary. In sensitivity analysis, 37 out of 48 (77.08%) miRNAs were consistently dysregulated. SIGNIFICANCE: A total of 48 dysregulated miRNAs were confirmed in this meta-analysis. MiR-122-5p and miR-133a-3p may be valuable biomarkers for CHD.

Latent class analysis to evaluate performance of plasma cortisol, plasma catecholamines, and SHSQ-25 for early recognition of suboptimal health status.

BACKGROUND: Chronic stress is associated with suboptimal health status (SHS) which is a new public health challenge in China and worldwide. Plasma stress hormones may act as potential objective biomarkers for SHS measure. This study was aimed to evaluate the diagnostic performance of plasma cortisol, catecholamine adrenaline/noradrenaline, and SHS questionnaires (SHSQ) for SHS using latent class analysis (LCA) in the absence of a gold standard. METHODS: A cross-sectional study was conducted among 868 employees in Beijing. The SHS questionnaires-25 (SHSQ-25) was distributed, and plasma cortisol, adrenaline, and noradrenaline were measured in the survey. LCA was used to assess the performance of both subjective and objective measures for SHS recognition. RESULTS: Akaike information criterion (AIC) and consistent AIC (CAIC) was 14.11 and 54.48 respectively, indicating that the model was well fitted. The sensitivity and specificity of plasma cortisol were 0.836 (95% CI 0.811-0.861) and 0.840 (95% CI 0.816-0.864), respectively. The area under curve (AUC) of receiver operating characteristic (ROC) of SHSQ-25 was 0.743 (95% CI 0.709-777), while the AUC of plasma adrenaline was 0.688 (95% CI 0.651-0.725). The prevalence of SHS in the investigated population was 34.78%. CONCLUSION: Plasma cortisol is a valuable biomarker for SHS detection, whereas SHSQ-25 is more suitable for SHS screening in the population-based health survey. The accuracy and applicability of plasma adrenaline are inferior to cortisol and SHSQ-25, respectively. LCA has merit to evaluate performance of plasma cortisol, catecholamines, and SHSQ-25 for recognition of SHS in the absence of a gold standard test.

Association analysis of copy number variations in type 2 diabetes-related susceptible genes in a Chinese population.

AIMS: Copy number variations (CNVs) have been implicated as an important genetic marker of common disease. In this study, we explored genetic effects of common CNVs in Type 2 diabetes (T2D) related susceptible genes in Chinese population. METHODS: Seven common CNV loci were selected from genes enclosing the susceptible single nucleotide polymorphisms (SNPs) of T2D confirmed by genome-wide association studies (GWAS) and replication studies conducted in east Asia population. The CNVs and SNPs were genotyped in 504 T2D patients and 494 non-T2D controls. Cumulative effect of the positive CNV loci was measured using genetic risk score (GRS). Multiplicative and additive interaction between candidate CNV loci and SNPs were assessed. RESULTS: Compared with the common two copies, the deletion of nsv6360 (adjusted OR = 2.28, 95% CI 1.37-3.78, P = 0.001), nsv8414 (adjusted OR = 1.89, 95% CI 1.16-3.08, P = 0.006) and nsv1898 (adjusted OR = 1.84, 95% CI 1.19-2.84, P = 0.005) were significantly associated with increased risk of T2D (P < 0.007). Significant dose-response relationship was observed between GRS and the risk of T2D (χ2 for trend = 19.51, P < 0.001). In addition, significant additive interactions between nsv8414 and rs17584499 in PTPRD (AP = 0.60, 95% CI 0.12-1.07) and nsv1898 and rs16955379 in CMIP (AP = 0.46, 95% CI 0.01-0.91) were observed. CONCLUSIONS: There were three CNV loci (nsv6360, nsv8414 and nsv1898) associated with T2D, and a significant cumulative effect of these loci on the risk of T2D. The comprehensive effects of both CNVs and SNPs may provide a more useful tool for the identification of genetic susceptibility for T2D.

Investigation of the Relationship Between Chronic Stress and Insulin Resistance in a Chinese Population.

BACKGROUND: Chronic stress may facilitate the development of metabolic diseases. Insulin resistance is present long before the clinical manifestations of individual metabolic abnormalities. To explore whether chronic stress is an independent risk factor of insulin resistance, we investigated the relationship between the stress system, selected parameters of energy homeostasis, and insulin resistance in a Chinese population. METHODS: We recruited 766 workers employed at four companies in Beijing. The degree of insulin resistance was determined using the homeostasis model assessment of insulin resistance (HOMA-IR). The highest quartile of HOMA-IR among all study subjects was further defined as insulin resistance in our study. The short standard version of the Copenhagen Psychosocial Questionnaire (COPSOQ) was used to assess job-related psychosocial stress. Pearson's correlation coefficients were calculated between cortisol level and HOMA-IR and components of metabolic syndrome, with stratification by gender. The relationship between cortisol and HOMA-IR independent of obesity was analyzed using a linear mixed model with company as a cluster unit. RESULTS: The values of the two scales of COPSOQ, including "demands at work" and "insecurity at work", were significantly associated with insulin resistance and cortisol concentration (P < 0.05). Cortisol was significantly positively correlated with glucose, HOMA-IR, and waist circumference in males and females (P < 0.05). After adjusting for potential confounders, cortisol was an independent positive predictor for HOMA-IR (P < 0.05). CONCLUSIONS: These findings showed that chronic stress was associated with insulin resistance and may contribute to the development of insulin resistance.