Ukr.Biochem.J. 2026; Volume 98, Issue 4, Jul-Aug, pp. 65-74

doi: https://doi.org/10.15407/ubj98.04.065

Estimation of circulating microRNAs as biomarkers in early stages of chronic kidney disease

H. H. Al-Shukri1*, R. S. Al-Azawi2, S. H. Ali2,

1College of Veterinary Medicine, Al-Qasim Green University,51013, Babylon, Iraq;
2College of Science, Al-Qasim Green University, 51013, Babylon, Iraq;
*e-mail: hamza14shukri72@gmail.com

Received: 29 April 2026; Revised: 24 June 2026;
Accepted: 27 July 2026; Available on-line: 04 August 2026

Background. Chronic kidney disease (CKD) is a progressive disorder that affects 10–13% of the worldwide population. The conventional biomarkers, like serum creatinine and estimated glomerular filtration rate (eGFR), have poor sensitivity in detecting early stage CKD, especially as it relates to Type 2 Diabetes Mellitus (T2DM). Recent studies suggest that circulating microRNAs (miRNAs) are sensitive molecular markers of renal pathology. Objectives. This study quantified the serum levels of miR-21, miR-155 and miR-192 in early-stage CKD patients with or without co-existing T2DM and determined the diagnostic performance of these microRNAs compared to cystatin C and eGFR. Methods. A case-control study was conducted with a total of 100 participants classified into three groups; healthy controls (n = 30), CKD patients with T2DM (n = 35) and CKD patients without T2DM (n = 35). The expression of serum miRNA was measured with real-time quantitative PCR using the 2⁻ΔΔCt method, U6 snRNA as internal reference. Cystatin C was determined by particle-enhanced immunonephelometry; eGFR was calculated using the CKD-EPI equation. Statistical analyses were performed using Kruskal-Wallis test, Spearman partial correlations, receiver operating characteristic (ROC) analysis, and multivariate logistic regression. Results. Significant upregulation of all three miRNAs in both CKD groups (P < 0.001) with a differing magnitude of fold-change compared with controls and the highest amplification for miR-192 was detected. ROC analysis identified miR-192 to be the best performing single biomarker (AUC = 0.937; 95% CI: 0.880–0.978). Spearman partial correlation networks showed strong positive relations of miRNAs with cystatin C (ρ = 0.74–0.80) and stronger negative associations with eGFR (ρ = −0.69 to −0.78). Conclusions. Circulating miR-21, miR-155 and miR-192 are promising minimally invasive biomarkers better than conventional markers for early CKD detection. Cystatin C was co-measured with it, yielding a highly accurate diagnostic panel especially in the diabetic nephropathy sub-phenotype. Clinical applicability requires validation in large multicentre cohorts.

Keywords: , , , , ,


References:

  1. Al Rashdi F, Tabche C, Atwan Z, Al-Qanubi H, Al Khaldi S, Al-Zadjali N, Rawaf S. Chronic kidney disease in hypertensive patients: the urgent need for targeted interventions in Arab countries: a systematic review. Front Nephrol. 2026;6:1735217. PubMed, PubMedCentral, CrossRef
  2. Kostev K, Lang M, Tröbs SO, Urbisch S, Gabler M. The Underdiagnosis of Chronic Kidney Disease in Patients with a Documented Estimated Glomerular Filtration Rate and/or Urine Albumin-Creatinine Ratio in Germany. Medicina (Kaunas). 2025;61(5):843. PubMed, PubMedCentral, CrossRef
  3. Alicic RZ, Rooney MT, Tuttle KR. Diabetic Kidney Disease: Challenges, Progress, and Possibilities. Clin J Am Soc Nephrol. 2017;12(12):2032-2045. PubMed, PubMedCentral, CrossRef
  4. Thomas HY, Ford Versypt AN. Pathophysiology of mesangial expansion in diabetic nephropathy: mesangial structure, glomerular biomechanics, and biochemical signaling and regulation.
    J Biol Eng. 2022;16(1):19. PubMed, PubMedCentral, CrossRef
  5. Neama ED, Al-Azawi RS, Kzar HH, Naji HH, Al-Alwany, EAH. Assessment of alpha-1-acid glycoprotein (AGP) and alpha-1-antitrypsin (AAT) in patients with type 2 diabetes mellitus. Ro J Med Pract. 2025;20(4):396.  CrossRef
  6. Obert LA, Elmore SA, Ennulat D, Frazier KS. A Review of Specific Biomarkers of Chronic Renal Injury and Their Potential Application in Nonclinical Safety Assessment Studies. Toxicol Pathol. 2021;49(5):996-1023. PubMed, PubMedCentral, CrossRef
  7. O’Brien J, Hayder H, Zayed Y, Peng C. Overview of MicroRNA Biogenesis, Mechanisms of Actions, and Circulation. Front Endocrinol (Lausanne). 2018;9:402. PubMed, PubMedCentral, CrossRef
  8. Lohajová Behulová R, Bugalová A, Bugala J, Struhárňanská E, Šafranek M, Juráš I. Circulating exosomal miRNAs as a promising diagnostic biomarker in cancer. Physiol Res. 2023;72(S3):S193-S207. PubMed, PubMedCentral, CrossRef
  9. Rodzoń-Norwicz M, Kogut P, Sowa-Kućma M, Gala-Błądzińska A. What a Modern Physician Should Know About microRNAs in the Diagnosis and Treatment of Diabetic Kidney Disease. Int J Mol Sci. 2025;26(14):6662. PubMed, PubMedCentral, CrossRef
  10. Kozomara A, Birgaoanu M, Griffiths-Jones S. miRBase: from microRNA sequences to function. Nucleic Acids Res. 2019;47(D1):D155-D162. PubMed, PubMedCentral, CrossRef
  11. Margaritis K, Margioula-Siarkou G, Giza S, Kotanidou EP, Tsinopoulou VR, Christoforidis A, Galli-Tsinopoulou A. Micro-RNA Implications in Type-1 Diabetes Mellitus: A Review of Literature. Int J Mol Sci. 2021;22(22):12165. PubMed, PubMedCentral, CrossRef
  12. Ren H, Wang Q. Non-Coding RNA and Diabetic Kidney Disease. DNA Cell Biol. 2021;40(4):553-567.
    PubMed, CrossRef
  13. Motshwari DD, Matshazi DM, Erasmus R, Kengne AP, Matsha TE, George C. MicroRNAs associated with chronic kidney disease in the general population and high-risk subgroups: protocol for a systematic review and meta-analysis. BMJ Open. 2022;12(2):e057500. PubMed, PubMedCentral, CrossRef
  14. AbdAli MR, Hamzah EF, Neama ED, Hadi SJ, Kzar HH, Al-Azawi RS, Kadhim ZH, Aboktifa MA, Ibrahim NJ, Jassim AM, Al-Gazally ME. (2023). Investigation of the PON1 (C> T rs705379) SNP and its correlation with physiological adiponectin levels in insulin resistance T2D patients induced by obesity. Int J Chem Biochem Sci. 2023;23(1):314-319.
  15. Zou Z, Zhou N, Zhang C. miRNA in the Progression of Diabetic Kidney Disease: New Insight. Int J Mol Sci. 2025;27(1):420. PubMed, PubMedCentral, CrossRef
  16. Rao X, Huang X, Zhou Z, Lin X. An improvement of the 2ˆ(-delta delta CT) method for quantitative real-time polymerase chain reaction data analysis. Biostat Bioinforma Biomath. 2013;3(3):71-85. PubMed, PubMedCentral
  17. Motawea M, Khalel MS, Kandil I, Faheem AM, Zaki MES, Abdelsalam M, Kyrillos F. Study of MicroRNA-192 as an Early Biomarker for Diagnosis of Diabetic Nephropathy. Diagnostics (Basel). 2025;15(12):1504. PubMed, PubMedCentral, CrossRef
  18. Wan X, Liao J, Lai H, Zhang S, Cui J, Chen C. Roles of microRNA-192 in diabetic nephropathy: the clinical applications and mechanisms of action. Front Endocrinol (Lausanne). 2023;14:1179161. PubMed, PubMedCentral, CrossRef
  19. Wang L, Wang HL, Liu TT, Lan HY. TGF-Beta as a Master Regulator of Diabetic Nephropathy. Int J Mol Sci. 2021;22(15):7881. PubMed, PubMedCentral, CrossRef
  20. Larrue R, Fellah S, Van der Hauwaert C, Hennino MF, Perrais M, Lionet A, Glowacki F, Pottier N, Cauffiez C. The Versatile Role of miR-21 in Renal Homeostasis and Diseases. Cells. 2022;11(21):3525. PubMed, PubMedCentral, CrossRef
  21. Pasca S, Jurj A, Petrushev B, Tomuleasa C, Matei D. MicroRNA-155 Implication in M1 Polarization and the Impact in Inflammatory Diseases. Front Immunol. 2020;11:625. PubMed, PubMedCentral, CrossRef
  22. Tsai CW, Grams ME, Inker LA, Coresh J, Selvin E. Cystatin C- and creatinine-based estimated glomerular filtration rate, vascular disease, and mortality in persons with diabetes in the U.S. Diabetes Care. 2014;37(4):1002-1008. PubMed, PubMedCentral, CrossRef
  23. Naji HH, Al-Azawi RSA, Ibrahim NJ, Kzar HH. Investigation of the Role of Zn/Cu Index and its Correlation with Physiological Activity of SOD 1 and GRx in Males with Acne Vulgaris. Arch Razi Inst. 2022;77(2):623-628. PubMed, PubMedCentral, CrossRef
  24. Kar S, Paglialunga S, Islam R. Cystatin C Is a More Reliable Biomarker for Determining eGFR to Support Drug Development Studies. J Clin Pharmacol. 2018;58(10):1239-1247. PubMed, CrossRef

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