Top

12-02-2024 | Diabetic Nephropathy | Original Article

Exosomal UMOD gene expression and urinary uromodulin level as early noninvasive diagnostic biomarkers for diabetic nephropathy in type 2 diabetic patients

Authors: Shaimaa I. Barr, Sahar S. Bessa, Tarek M. Mohamed, Eman M. Abd El-Azeem

Abstract

Background

Diabetic nephropathy (DN) is the leading cause of end-stage renal disease. Exosomes are promising biomarkers for disease diagnosis and uromodulin is a kidney-specific protein. So, this study was designed to investigate the change in the gene expression of urinary exosomal uromodulin mRNA and urinary uromodulin level and determine the diagnostic potential of these noninvasive biomarkers in the early stage of diabetic nephropathy in type 2 diabetic patients.

Method

This study included 100 participants; urinary exosomes were isolated using polyethylene glycol (PEG). Gene expression of exosomal uromodulin mRNA was determined by quantitative real-time polymerase chain reaction (q-RT-PCR). The urinary uromodulin levels were determined by an enzyme-linked immunosorbent assay (ELISA).

Result

In this study, the gene expression of exosomal uromodulin (UMOD) mRNA and the level of urinary uromodulin showed a significant increase in all diabetic groups with and without nephropathy compared to the control group. The exosomal UMOD mRNA showed a significant positive correlation with urinary uromodulin in all groups. Multiple logistic regression showed that urinary uromodulin was an independent determinant for DN. A diagnostic model of two indicators, exosomal UMOD mRNA and urinary uromodulin, can significantly predict DN. The area under the curve is 0.095, with a 95% confidence interval of 0.98–1, and 0.81, with a 95% confidence interval of 0.69–0.92, for the exosomal UMOD mRNA and urinary uromodulin, respectively.

Conclusion

Urinary exosomal mRNA of UMOD and urinary uromodulin levels are progressively elevated in an early stage of DN, even before the microalbuminuria stage, so they could be used as early predictors for DN.

Kim KS, Lee JS, Park JH, Lee EY, Moon JS, Lee SK, et al. Identification of novel biomarker for early detection of diabetic nephropathy. Biomedicines. 2021;9(5):457.CrossRefPubMedPubMedCentral

Mizdrak M, Kumrić M, Kurir TT, Božić J. Emerging biomarkers for early detection of chronic kidney disease. J Pers Med. 2022;12(4):548.CrossRefPubMedPubMedCentral

Oh SW, Kim S, Na KY, Chae DW, Kim S, Jin DC, et al. Clinical implications of pathologic diagnosis and classification for diabetic nephropathy. Diabetes Res Clin Pract. 2012;97(3):418–24. https://doi.org/10.1016/j.diabres.2012.03.016.CrossRefPubMed

Tachibana S, Iyoda M, Suzuki T, Kanazawa N, Honda H. Serum uromodulin levels reflect severity of clinicopathological findings in early stage IgA nephropathy. Am J Nephrol. 2022;53(7):575–85. https://doi.org/10.1159/000525836.CrossRefPubMed

Devuyst O, Bochud M, Olinger E. UMOD and the architecture of kidney disease. Pflügers Arch Eur J Physiol. 2022;474(8):771–81. https://doi.org/10.1007/s00424-022-02733-4.CrossRef

Renigunta A, Renigunta V, Saritas T, Decher N, Mutig K, Waldegger S. Tamm-Horsfall glycoprotein interacts with renal outer medullary potassium channel ROMK2 and regulates its function. J Biol Chem. 2011;286(3):2224–35.CrossRefPubMed

Wolf MT, Wu XR, Huang CL. Uromodulin upregulates TRPV5 by impairing caveolin-mediated endocytosis. Kidney Int. 2013;84(1):130–7.CrossRefPubMedPubMedCentral

Schmid M, Prajczer S, Gruber LN, Bertocchi C, Gandini R, Pfaller W, et al. Uromodulin facilitates neutrophil migration across renal epithelial monolayers. Cell Physiol Biochem. 2010;26(3):311–8.CrossRefPubMed

El-Achkar TM, McCracken R, Liu Y, Heitmeier MR, Bourgeois S, Ryerse J, et al. Tamm-Horsfall protein translocates to the basolateral domain of thick ascending limbs, interstitium, and circulation during recovery from acute kidney injury. Am J Physiol Renal Physiol. 2013;304(8):F1066–75. https://doi.org/10.1152/ajprenal.00543.2012.CrossRefPubMedPubMedCentral

10.

Boder P, Mary S, Mark PB, Leiper J, Dominiczak AF, Padmanabhan S, et al. Mechanistic interactions of uromodulin with the thick ascending limb: perspectives in physiology and hypertension. J Hypertens. 2021;39(8):1490–504. https://doi.org/10.1097/HJH.0000000000002861.CrossRefPubMedPubMedCentral

11.

Latifkar A, Hur YH, Sanchez JC, Cerione RA, Antonyak MA. New insights into extracellular vesicle biogenesis and function. J Cell Sci. 2019;132(13):jcs222406.CrossRefPubMedPubMedCentral

12.

Van Niel G, d’Angelo G, Raposo G. Shedding light on the cell biology of extracellular vesicles. Nat Rev Mol Cell Biol. 2018;19(4):213–28. https://doi.org/10.1038/nrm.2017.125.CrossRefPubMed

13.

Akers JC, Gonda D, Kim R, Carter BS, Chen CC. Biogenesis of extracellular vesicles (EV): exosomes, microvesicles, retrovirus-like vesicles, and apoptotic bodies. J Neurooncol. 2013;113(1):1–11. https://doi.org/10.1007/s11060-013-1084-8.CrossRefPubMedPubMedCentral

14.

Petrovčíková E, Vičíková K, Leksa V. Extracellular vesicles – biogenesis, composition, function, uptake and therapeutic applications. Biologia. 2018;73(4):437–48. https://doi.org/10.2478/s11756-018-0047-0.CrossRef

15.

Sinha N, Kumar V, Puri V, Nada R, Rastogi A, Jha V, et al. Urinary exosomes: potential biomarkers for diabetic nephropathy. Nephrology (Carlton). 2020;25(12):881–7. https://doi.org/10.1111/nep.13720.CrossRefPubMed

16.

Okamura A, Yoshioka Y, Saito Y, Ochiya T. Can extracellular vesicles as drug delivery systems be a game changer in cardiac disease? Pharm Res. 2022. https://doi.org/10.1007/s11095-022-03463-z.CrossRefPubMedPubMedCentral

17.

Ortega-Sanchez FG, Teresa V, Widmann T, Regiart M, Jerez-Salcedo MT, Fernández-Baldo MA, et al. Microfluidic systems in extracellular vesicles single analysis. A systematic review. TrAC Trends Anal Chem. 2023;159:116920.CrossRef

18.

Meng Y, Asghari M, Aslan MK, Yilmaz A, Mateescu B, Stavrakis S, et al. Microfluidics for extracellular vesicle separation and mimetic synthesis: recent advances and future perspectives. Chem Eng J. 2020;404:126110. https://doi.org/10.1016/j.cej.2020.126110.CrossRef

19.

Mateescu B, Kowal EJ, van Balkom BW, Bartel S, Bhattacharyya SN, Buzás EI, et al. Obstacles and opportunities in the functional analysis of extracellular vesicle RNA–an ISEV position paper. J Extracell Vesicles. 2017;6(1):1286095.CrossRefPubMedPubMedCentral

20.

Draznin B, Aroda VR, Bakris G, Benson G, Brown FM, American Diabetes Association Professional Practice C, et al. 11. Chronic kidney disease and risk management: Standards of Medical Care in Diabetes-2022. Diabetes Care. 2022;45(Suppl 1):S175–84.

21.

Lv CY, Ding WJ, Wang YL, Zhao ZY, Li JH, Chen Y, et al. A PEG-based method for the isolation of urinary exosomes and its application in renal fibrosis diagnostics using cargo miR-29c and miR-21 analysis. Int Urol Nephrol. 2018;50(5):973–82.CrossRefPubMed

22.

Weng Y, Sui Z, Shan Y, Hu Y, Chen Y, Zhang L, et al. Effective isolation of exosomes with polyethylene glycol from cell culture supernatant for in-depth proteome profiling. Analyst. 2016;141(15):4640–6. https://doi.org/10.1039/c6an00892e.CrossRefPubMed

23.

Yamamoto CM, Murakami T, Oakes ML, Mitsuhashi M, Kelly C, Henry RR, et al. Uromodulin mRNA from urinary extracellular vesicles correlate to kidney function decline in type 2 diabetes mellitus. Am J Nephrol. 2018;47(5):283–91. https://doi.org/10.1159/000489129.CrossRefPubMed

24.

Wang H, Zhang R, Wu X, Chen Y, Ji W, Wang J, et al. The Wnt signaling pathway in diabetic nephropathy. Front Cell Dev Biol. 2021;9:701547. https://doi.org/10.3389/fcell.2021.701547.CrossRefPubMed

25.

Erekat NS. Programmed cell death in diabetic nephropathy: a review of apoptosis autophagy and necroptosis. Med Sci Monit Int Med J Exp Clin Res. 2022;28:e937766-e.

26.

Xie S, Zhang Q, Jiang L. Current knowledge on exosome biogenesis, Cargo-sorting mechanism and therapeutic implications. Membranes. 2022;12(5):498.CrossRefPubMedPubMedCentral

27.

Zhang H, Zhang Q, Deng Y, Chen M, Yang C. Improving isolation of extracellular vesicles by utilizing nanomaterials. Membranes (Basel). 2021;12(1):55.CrossRefPubMed

28.

Micanovic R, LaFavers K, Garimella PS, Wu X-R, El-Achkar TM. Uromodulin (Tamm–Horsfall protein): guardian of urinary and systemic homeostasis. Nephrol Dial Transplant. 2020;35(1):33–43.CrossRefPubMed

29.

Säemann MD, Weichhart T, Zeyda M, Staffler G, Schunn M, Stuhlmeier KM, et al. Tamm-Horsfall glycoprotein links innate immune cell activation with adaptive immunity via a Toll-like receptor-4–dependent mechanism. J Clin Investig. 2005;115(2):468–75.CrossRefPubMedPubMedCentral

30.

El-Achkar TM, Dagher PC. Tubular cross talk in acute kidney injury: a story of sense and sensibility. Am J Physiol Renal Physiol. 2015;308(12):F1317–23.CrossRefPubMedPubMedCentral

31.

Liu Y, El-Achkar TM, Wu X-R. Tamm-Horsfall protein regulates circulating and renal cytokines by affecting glomerular filtration rate and acting as a urinary cytokine trap. J Biol Chem. 2012;287(20):16365–78. https://doi.org/10.1074/jbc.M112.348243.CrossRefPubMedPubMedCentral

32.

Rhodes DC. Importance of carbohydrate in the interaction of Tamm-Horsfall protein with complement 1q and inhibition of classical complement activation. Immunol Cell Biol. 2006;84(4):357–65. https://doi.org/10.1111/j.1440-1711.2006.01434.x.CrossRefPubMed

33.

LaFavers KA, Macedo E, Garimella PS, Lima C, Khan S, Myslinski J, et al. Circulating uromodulin inhibits systemic oxidative stress by inactivating the TRPM2 channel. Sci Transl Med. 2019;11(512):eaaw3639.CrossRefPubMedPubMedCentral

34.

Srivastava R, Micanovic R, El-Achkar TM, Janga SC. An intricate network of conserved DNA upstream motifs and associated transcription factors regulate the expression of uromodulin gene. J Urol. 2014;192(3):981–9. https://doi.org/10.1016/j.juro.2014.02.095.CrossRefPubMed

35.

Lou NJ, Ni YH, Jia HY, Deng JT, Jiang L, Zheng FJ, et al. Urinary Microvesicle-bound uromodulin: a potential molecular biomarker in diabetic kidney disease. J Diabetes Res. 2017. https://doi.org/10.1155/2017/3918681.CrossRefPubMedPubMedCentral

36.

Köttgen A, Hwang SJ, Larson MG, Van Eyk JE, Fu Q, Benjamin EJ, et al. Uromodulin levels associate with a common UMOD variant and risk for incident CKD. J Am Soc Nephrol. 2010;21(2):337–44. https://doi.org/10.1681/asn.2009070725.CrossRefPubMedPubMedCentral

37.

El-Achkar TM, Wu XR. Uromodulin in kidney injury: an instigator, bystander, or protector? Am J Kidney Dis. 2012;59(3):452–61. https://doi.org/10.1053/j.ajkd.2011.10.054.CrossRefPubMedPubMedCentral

38.

Chakraborty J, Below AA, Solaiman D. Tamm-Horsfall protein in patients with kidney damage and diabetes. Urol Res. 2004;32(2):79–83. https://doi.org/10.1007/s00240-003-0374-6.CrossRefPubMed

Title: Exosomal UMOD gene expression and urinary uromodulin level as early noninvasive diagnostic biomarkers for diabetic nephropathy in type 2 diabetic patients
Authors: Shaimaa I. Barr
Sahar S. Bessa
Tarek M. Mohamed
Eman M. Abd El-Azeem
Publication date: 12-02-2024
Publisher: Springer Nature Singapore
Keywords: Diabetic Nephropathy
Microalbuminuria
Biomarkers
Macroalbuminuria
Published in: Diabetology International
Print ISSN: 2190-1678
Electronic ISSN: 2190-1686
DOI: https://doi.org/10.1007/s13340-023-00686-2

Live Webinar | 27-06-2024 | 18:00 (CEST)

Keynote webinar | Spotlight on medication adherence

Reserve your place

Live: Thursday 27th June 2024, 18:00-19:30 (CEST)

WHO estimates that half of all patients worldwide are non-adherent to their prescribed medication. The consequences of poor adherence can be catastrophic, on both the individual and population level.

Join our expert panel to discover why you need to understand the drivers of non-adherence in your patients, and how you can optimize medication adherence in your clinics to drastically improve patient outcomes.

Prof. Kevin Dolgin

Prof. Florian Limbourg

Prof. Anoop Chauhan

Developed by: Springer Medicine

Obesity Clinical Trial Summary

At a glance: The STEP trials

A round-up of the STEP phase 3 clinical trials evaluating semaglutide for weight loss in people with overweight or obesity.

Developed by: Springer Medicine

Highlights from the ACC 2024 Congress

Year in Review: Pediatric cardiology

Pediatric Cardiology Video

Watch Dr. Anne Marie Valente present the last year's highlights in pediatric and congenital heart disease in the official ACC.24 Year in Review session.

Year in Review: Pulmonary vascular disease

Cardiopulmonary Comorbidity Video

The last year's highlights in pulmonary vascular disease are presented by Dr. Jane Leopold in this official video from ACC.24.

Year in Review: Valvular heart disease

Valvular Heart Disease Video

Watch Prof. William Zoghbi present the last year's highlights in valvular heart disease from the official ACC.24 Year in Review session.

Year in Review: Heart failure and cardiomyopathies

Heart Failure Video

Watch this official video from ACC.24. Dr. Biykem Bozkurt discusses last year's major advances in heart failure and cardiomyopathies.

ACC 2024 Congress Coverage

Year in Review: Heart failure and cardiomyopathies

Heart Failure Video

Watch this official video from ACC.24. Dr. Biykem Bozkurt discusses last year's major advances in heart failure and cardiomyopathies.

Watch now

Semaglutide benefits people with type 2 diabetes and obesity-related heart failure

16-04-2024 Type 2 Diabetes News

Incentivised to exercise

11-04-2024 Lifestyle Modifications News

New intervention for STEMI-related cardiogenic shock

10-04-2024 Myocardial Infarction News

» View more highlights on the ACC 2024 congress hub page

Image Credits

Illustration of figure on mountain summit/© Orapun / Stock.adobe.com, STEP AAG HeroTeaserCollection image/© Tarek / Generated with AI / Stock.adobe.com, ACC 2024 Pediatric and Congenital Heart Disease: Year in Review/© 2024 American College of Cardiology, ACC 2024 Pulmonary Vascular Disease: Year in Review/© 2024 American College of Cardiology, ACC 2024 Valvular Heart Disease: Year in Review/© 2024 American College of Cardiology, ACC 2024 HF and Cardiomyopathies: Year in Review/© 2024 American College of Cardiology, Woman out walking/© Seventyfour / stock.adobe.com (symbolic image with model), Woman checking smartwatch /© saltdium / stock.adobe.com (symbolic image with model), Cardiac catheterization/© Damian / stock.adobe.com