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Biomarkers in Diabetes Mellitus: From Discovery to Clinical Application

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A Special Issue of International Journal of Molecular Sciences (ISSN 1422-0067) belonging to the section "Molecular Pathology, Diagnostics, and Therapeutics".

Deadline for manuscript submissions: 25 December 2026 | Viewed by 11110

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Prof. Dr. Jelena Vekic

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Department of Medical Biochemistry, Faculty of Pharmacy, University of Belgrade, 11000 Belgrade, Serbia
Interests: lipids; lipoproteins; atherosclerosis; diabetes; biomarkers; laboratory diagnostics; medical biochemistry
Special Issues, Collections and Topics in MDPI journals

Prof. Dr. Aleksandra Zeljkovic

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Department of Medical Biochemistry, Faculty of Pharmacy, University of Belgrade, 11000 Belgrade, Serbia
Interests: lipoprotein metabolism; cholesterol homeostasis; non-cholesterol sterols; sphingolipids; lipid biomarkers; dyslipidemia in pregnancy
Special Issues, Collections and Topics in MDPI journals

Special Issue Information

Dear Colleagues,

Diabetes mellitus remains a significant global health challenge, characterized by rising prevalence and a substantial healthcare burden due to associated complications. Given the heterogeneous nature and multifactorial pathogenesis of diabetes, there is a critical need for accurate diagnostic and prognostic tools to improve patient outcomes. In this context, biomarkers play a pivotal role by enabling early detection, risk stratification, personalized therapeutic strategies and effective disease progression monitoring and treatment responses.

This Special Issue aims to highlight the latest advances in the discovery, validation and clinical translation of biomarkers associated with diabetes and its complications. We invite submissions of original research articles and review papers on genetic and biochemical biomarkers, particularly those that reflect the distinct molecular profiles of type 1, type 2 and gestational diabetes. We especially encourage contributions focused on predictive and prognostic biomarkers of microvascular and macrovascular diabetes complications. Submissions employing advanced molecular approaches, including lipidomics, transcriptomics, metabolomics and integrated multi-omics strategies, are highly welcome. We are also interested in translational and clinical research that explores the application of biomarkers in real-world healthcare settings, especially those that inform therapeutic decision making, as well as studies that support the integration of molecular biomarkers into precision medicine frameworks. We look forward to your valuable contributions.

Prof. Dr. Jelena Vekic
Prof. Dr. Aleksandra Zeljkovic
Guest Editors

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Keywords

diabetes mellitus
biomarkers
diabetes complications
omics technologies
risk stratification
therapeutic response
personalized medicine

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Published Papers (8 papers)

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Research

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19 pages, 3972 KB

Open AccessArticle

Microvesicle-Derived Redox Signatures as Mediators of Endothelial Dysfunction in Diabetes

by Sarah Khalaf Ghanem, Hanan H. Abunada, Shahenda Salah Abdelsalam, Loulia Bader and Abdelali Agouni

Int. J. Mol. Sci. 2026, 27(13), 6005; https://doi.org/10.3390/ijms27136005 - 4 Jul 2026

Viewed by 434

Abstract

Chronic hyperglycemia and excessive reactive oxygen species (ROS) production are defining features of endothelial dysfunction, a key driver of diabetic vascular complications such as diabetic nephropathy. Microvesicles (MV-enriched fraction), a subtype of extracellular vesicles, and the stress-responsive antioxidant protein Sestrin2 (SESN2) have emerged as important contributors to these processes. This study investigated the role of the MV-enriched fraction in endothelial cell communication under diabetic conditions, with a particular focus on oxidative stress signaling. To model diabetic injury, EA.hy926 endothelial cells were treated with methylglyoxal (MGO), and the resulting MV-enriched fraction was isolated and then applied to two recipient models: naïve endothelial cells and SESN2 knockdown (KD) cells. Protein expression of key antioxidant markers, including endothelial nitric oxide synthase (eNOS), was assessed by Western blot. Nitric oxide (NO) bioavailability was quantified via nitrite measurement using 2,3-diaminonaphthalene (DAN), while mitochondrial and cytosolic ROS levels were evaluated using MitoSOX and dihydroethidium (DHE), respectively. Results demonstrated that the MV-enriched fraction derived from diabetic conditions triggers a complex antioxidant response in healthy endothelial cells, characterized by upregulation of SESN2, superoxide dismutase 1 (SOD1), and heme oxygenase-1 (HO-1). This suggests a compensatory mechanism that mitigates oxidative stress. Notably, SESN2 KD cells exhibited increased ROS production and reduced NO levels upon MV treatment, underscoring the essential role of SESN2 in maintaining redox homeostasis. Overall, this study highlights the dual role of the MV-enriched fraction as a mediator of both protective and detrimental redox signaling in diabetic endothelial dysfunction and suggests potential therapeutic targets for managing diabetic vascular complications. Full article

(This article belongs to the Special Issue Biomarkers in Diabetes Mellitus: From Discovery to Clinical Application)

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21 pages, 10454 KB

Open AccessArticle

Experimental Diabetic Retinopathy in Wistar Rats Induced by Streptozotocin: A Window into Retinal Disease Progression

by Luis Fernando Barba-Gallardo, Manuel Enrique Ávila-Blanco, Javier Ventura-Juárez, Martín Humberto Muñoz-Ortega, Ruth Clarisa Murillo-Ruíz, Marcela Rivera-Delgadillo, José Luis Díaz-Rubio, Elizabeth Casillas-Casillas, Luis Héctor Salas-Hernández and Paloma Lucía Guerra-Ávila

Int. J. Mol. Sci. 2026, 27(8), 3427; https://doi.org/10.3390/ijms27083427 - 11 Apr 2026

Viewed by 1261

Abstract

Diabetic retinopathy (DR), recognized as a progressive neurovascular and microvascular complication of diabetes, remains one of the leading causes of visual disability worldwide, within the context of a sustained increase in ophthalmic diseases and retinal vascular disorders that compromise vision. This study aimed to characterize the progression of diabetic retinopathy in a streptozotocin (STZ)-induced Wistar rat model. A single dose of 65 mg/kg body weight was administered, with follow-up periods at 2, 4, 8, and 10 weeks, compared to healthy controls. STZ-induced rats exhibited reduced weight gain compared to the control group. They also showed markedly variable hyperglycemia, with glucose concentrations ranging from 250 to 530 mg/dL. Histological analysis of retinal tissue at week 4 revealed early signs of vascular compromise, including early indications of a microenvironment conducive to neovascularization and edema. By week 8, retinal damage had progressed to hemorrhage, persistent edema, and layer-specific vascular disruption. At week 10, intensified neovascularization and exacerbated edema indicated advanced microvascular deterioration. Immunofluorescence analysis demonstrated a temporal accumulation of CD8⁺ T cells in the retina, correlating with photoreceptor degeneration. The coordinated dynamics of CD4⁺ and CD8⁺ T cells suggested transient immune activation during STZ-induced retinal degeneration. Gene expression profiling revealed a proinflammatory and pro-oxidative retinal microenvironment, characterized by the overexpression of angiogenic pathways and proliferative signals. Simultaneously, the antioxidant response appeared partially impaired. Collectively, these findings provide mechanistic perspective on the multifactorial nature of diabetic retinopathy. Oxidative stress, inflammation, and angiogenesis converge to disrupt retinal homeostasis. This experimental model may serve as a reliable platform for future studies aimed at elucidating disease pathophysiology, identifying novel therapeutic targets, and evaluating emerging ophthalmic antidiabetic interventions. Full article

(This article belongs to the Special Issue Biomarkers in Diabetes Mellitus: From Discovery to Clinical Application)

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12 pages, 736 KB

Open AccessArticle

Glycated Albumin and Cardiovascular Mortality in CKD Stage V Patients with Diabetes Mellitus: A Five-Year Follow-Up Study

by Ana Bulatovic, Nada Dimkovic, Svetlana Jelic, Aleksandar Jankovic, Tatjana Damjanovic, Verica Todorov-Sakic, Jelena Bjedov, Bojan Stopic, Petar Djuric and Radomir Naumovic

Int. J. Mol. Sci. 2026, 27(5), 2215; https://doi.org/10.3390/ijms27052215 - 26 Feb 2026

Cited by 1 | Viewed by 626

Abstract

Glycemic assessment in patients with chronic kidney disease (CKD) Stage V on hemodialysis (HD) is limited by the inaccuracy of hemoglobin A1c (HbA1c), mainly due to anemia, shortened erythrocyte lifespan, and erythropoiesis-stimulating agent (ESA) therapy. Glycated albumin (GA), independent of erythrocyte turnover, may better reflect glycemic exposure. We evaluated the diagnostic performance and clinical utility of GA as a biomarker of poor glycemic control and cardiovascular risk in patients with diabetes mellitus (DM). A cross-sectional analysis and five-year prospective follow-up were conducted in three subgroups: HD+DM+ (n = 40), HD- DM+ (n = 15), and HD+DM– (n = 22). Glycemic markers (mean plasma glucose over 3 months (PG₃m), GA, and HbA1c) were compared between groups. GA levels were significantly higher in HD+DM+ patients (p < 0.001) and showed the strongest correlation with PG₃m. GA independently predicted poor glycemic control (OR 3.23; 95% CI 1.54–6.77; p = 0.002) and demonstrated high diagnostic accuracy (AUC 0.873; optimal cut-off 10%). During the five-year follow-up, CV mortality was 35%, and 85% of deceased patients had GA > 10%. Although Cox regression did not reach statistical significance, GA showed a consistent trend toward higher CV mortality risk after adjustment for age and HD duration (HR 2.56; 95% CI 0.57–11.44; p = 0.21), whereas HbA1c was not prognostic (HR 1.39; 95% CI 0.49–3.91; p = 0.28). GA appears to be a clinically useful marker of glycemic control in diabetic patients with CKD Stage V receiving maintenance hemodialysis. In this cohort, GA showed high diagnostic accuracy and a more balanced sensitivity/specificity profile compared with HbA1c, with a consistent trend toward association with cardiovascular mortality. In this regard, the wider use of this marker in clinical practice is worth considering. Nevertheless, larger prospective studies are warranted to validate these observations. Full article

(This article belongs to the Special Issue Biomarkers in Diabetes Mellitus: From Discovery to Clinical Application)

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12 pages, 1002 KB

Open AccessArticle

Real-World Utility of GWAS-Based Diabetes Mellitus Panel Testing

by In Hwa Jeong, Kyung-Won Hong, Ja-Eun Choi and Bo-Kyung Shine

Int. J. Mol. Sci. 2026, 27(1), 275; https://doi.org/10.3390/ijms27010275 - 26 Dec 2025

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Abstract

This study evaluated the clinical utility of a polygenic risk score (PRS)-based multigene panel test for predicting diabetes mellitus (DM) in a healthy population. A total of 302 individuals underwent genetic testing using the HelloGene™ DM panel, which includes four DM-related single nucleotide polymorphisms (CDKAL1, HHEX, KCNQ1, and TCF7L2). PRS values were calculated using an algorithm developed from the Korean Genome and Epidemiology Study (KoGES; n = 39,605), and participants were classified into four genetic risk groups (low, moderate, high, and very high). Fasting blood glucose, glycated hemoglobin (HbA1c), and body mass index were assessed at baseline and after at least three years of follow-up, and lifestyle factors including smoking, alcohol consumption, and exercise status were recorded. No significant differences in age, sex, or lifestyle habits were observed among PRS groups. The very high-risk group showed significantly higher follow-up fasting blood glucose levels (p = 0.001) and higher baseline and follow-up HbA1c levels (p = 0.0025 and p = 0.001, respectively), as well as a 4.5-fold increased risk of developing DM compared with other groups. Smoking significantly modified genetic risk, with smokers in the very high-risk group showing a 25% higher likelihood of developing DM. CDKAL1 and TCF7L2 variants were most prevalent in the moderate- and high-risk groups, while HHEX variants in the high-risk group showed the greatest susceptibility, particularly among current smokers. Overall, PRS-based genetic testing demonstrated potential clinical utility for stratifying individuals according to relative diabetes risk, highlighting a possible interaction between genetic susceptibility and lifestyle factors such as smoking Full article

(This article belongs to the Special Issue Biomarkers in Diabetes Mellitus: From Discovery to Clinical Application)

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17 pages, 1752 KB

Open AccessArticle

Frequency of Polymorphisms in SLC47A1 (rs2252281 and rs2289669) and SLC47A2 (rs34834489 and rs12943590) and the Influence of SLC22A1 (rs72552763 and rs622342) on HbA1c Levels in Mexican-Mestizo Patients with DMT2 Treated with Metformin Monotherapy

by Milton Abraham Gómez-Hernández, Adiel Ortega-Ayala, Oscar Rodríguez-Lima, Abraham Landa, Gustavo Acosta-Altamirano and Juan A. Molina-Guarneros

Int. J. Mol. Sci. 2025, 26(17), 8652; https://doi.org/10.3390/ijms26178652 - 5 Sep 2025

Cited by 2 | Viewed by 1754

Abstract

Diabetes type 2 (DT2) entails significant health, economic, and productivity repercussions around the world. Poor glycaemic control, defined as an HbA1c >7.0%, has been associated with a number of complications. In spite of the large share of healthcare resources allocated to DT2 treatment, the proportion of controlled Mexican patients is among the lowest in the world (34.4%). Certain protein-encoding genetic polymorphisms involved in metformin transport may affect glycaemic control. We focused on determining the frequency of rs2289669, rs2252281, rs12943590, and rs34834489 polymorphisms in Mexican-Mestizo patients from the Tertiary Care Regional Hospital of Ixtapaluca, State of Mexico, Mexico, as well as assessing their possible association with therapeutic efficacy, as estimated through glycated haemoglobin. The individual polymorphism analysis did not reveal an association with glycaemic control; however, when combined with rs72552763 and rs622342, we found a significant positive correlation between HbA1c levels and metformin dose, which prevailed among patients carrying allelic variants of rs2289669 or rs12943590 who were also simultaneously carrying allelic variants of rs72552763 or rs622342. Patients carrying the reference allele of rs34834489 reported a significant positive correlation between HbA1c levels and metformin dose as well, regardless of their rs72552763 or rs622342 genotype. Thus, we identified alleles and allelic combinations of SLC47A1, SLC47A2, and SLC22A1 polymorphisms posing a potential glycaemic control risk in Mexican-Mestizo patients. Full article

(This article belongs to the Special Issue Biomarkers in Diabetes Mellitus: From Discovery to Clinical Application)

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18 pages, 389 KB

Open AccessArticle

Global DNA Methylation in Poorly Controlled Type 2 Diabetes Mellitus: Association with Redox and Inflammatory Biomarkers

by Sanja Vujcic, Jelena Kotur-Stevuljevic, Zoran Vujcic, Sanja Stojanovic, Teodora Beljic Zivkovic, Miljanka Vuksanovic, Milica Marjanovic Petkovic, Iva Perovic Blagojevic, Branka Koprivica-Uzelac, Sanja Ilic-Mijailovic, Manfredi Rizzo, Aleksandra Zeljkovic, Tatjana Stefanovic, Srecko Bosic and Jelena Vekic

Int. J. Mol. Sci. 2025, 26(14), 6716; https://doi.org/10.3390/ijms26146716 - 13 Jul 2025

Cited by 4 | Viewed by 2003

Abstract

Although emerging evidence suggests that epigenetic mechanisms contribute to the pathogenesis and progression of type 2 diabetes mellitus (T2DM), data remain limited for patients with suboptimal metabolic control. The aim of this study was to assess global DNA methylation in patients with poorly controlled T2DM and to identify diabetes-related factors associated with DNA methylation levels. The study included 107 patients and 50 healthy controls. Global DNA methylation (5mC) was measured by UHPLC-DAD method. Pro-oxidant and antioxidant biomarkers, advanced glycation end-products, high-sensitivity C-reactive protein (hsCRP) and complete blood count were determined and leukocyte indices calculated. Patients had a significantly lower 5mC than controls (3.56 ± 0.31% vs. 4.00 ± 0.68%; p < 0.001), with further reductions observed in those with longer disease duration and diabetic foot ulcers. Oxidative stress and inflammatory biomarkers were higher in the patient group. DNA hypomethylation was associated with a higher monocyte-to-lymphocyte ratio and hsCRP, pro-oxidant–antioxidant balance, ischemia-modified albumin, and advanced oxidation protein products levels. Conversely, 5mC levels showed positive correlations with total antioxidant status and total sulfhydryl groups. Principal component analysis identified five key factors: proinflammatory, pro-oxidant, aging, hyperglycemic, and antioxidant. The pro-oxidant factor emerged as the sole independent predictor of global DNA hypomethylation in T2DM (OR = 2.294; p = 0.027). Our results indicate that global DNA hypomethylation could be a biomarker of T2DM progression, reflecting the complex interactions between oxidative stress, inflammation, and epigenetic modifications in T2DM. Full article

(This article belongs to the Special Issue Biomarkers in Diabetes Mellitus: From Discovery to Clinical Application)

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Review

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23 pages, 5308 KB

Open AccessReview

Circulating CTRP1 in Adults with Type 2 Diabetes Mellitus: A Systematic Review and Meta-Analysis

by Anirudh Suresh, Alexander Dallaway, Attia Mustafa, Lukasz Lagojda, Chris Kite, Ioannis Kyrou and Harpal S. Randeva

Int. J. Mol. Sci. 2026, 27(15), 6775; https://doi.org/10.3390/ijms27156775 - 29 Jul 2026

Viewed by 444

Abstract

Type 2 diabetes mellitus (T2DM) is typically associated with a dysregulated adipokine profile. C1q tumour necrosis factor-related protein 1 (CTRP1) is a relatively novel adipokine with protective metabolic effects, including improved insulin sensitivity. Clinical studies have increasingly highlighted a potential association between circulating CTRP1 and T2DM. Therefore, this systematic review and meta-analysis aimed to synthesise existing data on the association between circulating CTRP1 concentrations and T2DM in adults. MEDLINE, EMBASE, Cochrane Library, Web of Science, CINAHL and PsycInfo databases were searched for studies measuring circulating CTRP1 in adults with T2DM and controls. Eligible data were meta-analysed, reporting the mean difference (MD) and 95% confidence intervals (CIs) of circulating CTRP1 concentrations between T2DM cases and controls. The Revised Risk of Bias Assessment Tool for Nonrandomized Studies of Interventions (RoBANS 2) and the National Institutes of Health (NIH) quality assessment tool were utilised for study risk of bias and quality assessments, respectively. Eight studies (525 T2DM cases; 612 non-T2DM controls) were eligible for meta-analysis, showing significantly higher circulating CTRP1 concentrations in T2DM cases than controls (MD: 185.31 ng/mL; 95% CI: 140.02 to 230.60; p < 0.001). Most included studies were cross-sectional, and considerable heterogeneity was noted, with five studies having a high risk of bias in at least one RoBANS 2 domain. The MD remained relatively unchanged following a leave-one-out sensitivity analysis and exclusion of poor-quality studies. Meta-regression indicated that studies with a greater proportion of males (β = −188.68, p = 0.01) and higher mean body mass index (β = −23.96, p = 0.02) were associated with a lower MD. These findings indicate that circulating CTRP1 concentrations are significantly higher in adults with T2DM compared to non-T2DM controls. Future prospective and large-scale studies are required to delineate the nature of this association and explore the potential of CTRP1 as a biomarker for T2DM. PROSPERO registration ID: CRD420251137104. Full article

(This article belongs to the Special Issue Biomarkers in Diabetes Mellitus: From Discovery to Clinical Application)

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28 pages, 1192 KB

Open AccessReview

From Molecules to Biomarkers: Nogo Proteins and Receptors in the Early Detection of Type 2 Diabetes Complications: A Systematic Review

by Jelena M. Bogdanović, Ivana Babić, Jelena Stanarčić Gajović, Sandra Singh Lukač, Dragana Mijač, Dušan Popović, Ivan Ranković, Ljiljana Popović, Iva Rasulić and Katarina Lalić

Int. J. Mol. Sci. 2026, 27(11), 5124; https://doi.org/10.3390/ijms27115124 - 5 Jun 2026

Cited by 2 | Viewed by 920

Abstract

Nogo (RTN4) proteins and their receptors have emerged as candidate mediators of metabolic regulation and vascular pathology relevant to type 2 diabetes (T2D). The primary objective of this PRISMA-guided systematic review was to evaluate the clinical and cohort evidence for RTN4/RTN4R as potential biomarkers of T2D progression and vascular complications. A secondary objective was to synthesize preclinical mechanistic evidence on the effects of Nogo axis modulation on pathways relevant to the pathogenesis of T2D. We performed a PRISMA-guided systematic review. The protocol was not prospectively registered in PROSPERO. To ensure reproducibility, we provide complete search keywords, the screening log and the full-text exclusion table. PubMed/MEDLINE, EMBASE and Web of Science were searched for studies published 2000–2025; full search keywords are provided in the main text. The search strategy combined and free-text terms with Boolean operators. We included original preclinical and clinical studies, cohort/proteomic analyses, meta-analyses, and mechanistic papers reporting expression, function, signaling, or clinical associations of Nogo proteins/receptors in metabolic or vascular outcomes. Exclusion criteria: non-English articles, unclear methods, studies outside 2000–2025, and studies lacking primary data. Two reviewers independently screened records; conflicts were resolved by consensus. Study quality was appraised using established tools (SYRCLE for animal studies, Newcastle–Ottawa Scale for cohort/case-control studies). Preclinical evidence supports tissue-specific roles for RTN4 isoforms and receptors in the regulation of insulin secretion, proGCG → GLP-1 processing, ER homeostasis, and vascular permeability through the Src/PI3K/Akt and RhoA/ROCK axes. Cohort and proteomic analyses report associations between RTN4/RTN4R or serum NogoB and faster progression of T2D or vascular complications, but genetic assessment of causality (Mendelian randomization) has so far provided limited support in available data sets. Findings are heterogeneous with respect to directionality and tissue localization. RTN4 signaling exhibits tissue-specific mechanisms relevant to glucose regulation and vascular biology and warrants further translational study. However, heterogeneity across studies and limited genetic support for causality indicate that isoform-specific quantitative validation, longitudinal cohorts and integrated genetic–functional analyses are required before RTN4/RTN4R can be considered as clinical biomarkers. Full article

(This article belongs to the Special Issue Biomarkers in Diabetes Mellitus: From Discovery to Clinical Application)

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