5-Hydroxymethylcytosines in Circulating Cell-Free DNA Reveal Vascular Complications of Type 2 Diabetes

Overview

Journal Clin Chem

Publisher Oxford University Press

Specialty Biochemistry

Date 2019 Oct 3

PMID 31575611

Citations 16

Authors

Ying Yang

Chang Zeng

Xingyu Lu

Yanqun Song

Ji Nie

Ruoxi Ran

Zhou Zhang

Chuan He

Wei Zhang

Song-Mei Liu

Affiliations

Soon will be listed here.

Abstract

Background: Long-term complications of type 2 diabetes (T2D), such as macrovascular and microvascular events, are the major causes for T2D-related disability and mortality. A clinically convenient, noninvasive approach for monitoring the development of these complications would improve the overall life quality of patients with T2D and help reduce healthcare burden through preventive interventions.

Methods: A selective chemical labeling strategy for 5-hydroxymethylcytosines (5hmC-Seal) was used to profile genome-wide 5hmCs, an emerging class of epigenetic markers implicated in complex diseases including diabetes, in circulating cell-free DNA (cfDNA) from a collection of Chinese patients (n = 62). Differentially modified 5hmC markers between patients with T2D with and without macrovascular/microvascular complications were analyzed under a case-control design.

Results: Statistically significant changes in 5hmC markers were associated with T2D-related macrovascular/microvascular complications, involving genes and pathways relevant to vascular biology and diabetes, including insulin resistance and inflammation. A 16-gene 5hmC marker panel accurately distinguished patients with vascular complications from those without [testing set: area under the curve (AUC) = 0.85; 95% CI, 0.73-0.96], outperforming conventional clinical variables such as urinary albumin. In addition, a separate 13-gene 5hmC marker panel could distinguish patients with single complications from those with multiple complications (testing set: AUC = 0.84; 95% CI, 0.68-0.99), showing superiority over conventional clinical variables.

Conclusions: The 5hmC markers in cfDNA reflected the epigenetic changes in patients with T2D who developed macrovascular/microvascular complications. The 5hmC-Seal assay has the potential to be a clinically convenient, noninvasive approach that can be applied in the clinic to monitor the presence and severity of diabetic vascular complications.

Citing Articles

5-hydroxymethylcytosine features of portal venous blood predict metachronous liver metastases of colorectal cancer and reveal phosphodiesterase 4 as a therapeutic target.

Xu N, Gao Z, Wu D, Chen H, Zhang Z, Zhang L Clin Transl Med. 2025; 15(2):e70189.

PMID: 39956959 PMC: 11830572. DOI: 10.1002/ctm2.70189.

Review of Associations of Diabetes and Insulin Resistance With Brain Health in Three Harmonised Cohort Studies of Ageing and Dementia.

Biswas R, Capuano A, Mehta R, Barnes L, Bennett D, Arvanitakis Z Diabetes Metab Res Rev. 2025; 41(1):e70032.

PMID: 39873127 PMC: 11774135. DOI: 10.1002/dmrr.70032.

Circulating Cell-Free DNA in Metabolic Diseases.

Pollastri A, Kovacs P, Keller M J Endocr Soc. 2025; 9(2):bvaf006.

PMID: 39850787 PMC: 11756337. DOI: 10.1210/jendso/bvaf006.

Literary Identification of Differentially Hydroxymethylated DNA Regions for Type 2 Diabetes Mellitus: A Scoping Minireview.

Luong R, Guan W, Vue F, Dai J Int J Environ Res Public Health. 2024; 21(2).

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Analysis of genome-wide 5-hydroxymethylation of blood samples stored in different anticoagulants: opportunities for the expansion of clinical resources for epigenetic research.

Gao L, Zhang Z, Cui X, West-Szymanski D, Ye C, He C Epigenetics. 2023; 18(1):2271692.

PMID: 37898998 PMC: 10627064. DOI: 10.1080/15592294.2023.2271692.

References

Parikh R, Mathai A, Parikh S, Sekhar G, Thomas R . Understanding and using sensitivity, specificity and predictive values. Indian J Ophthalmol. 2007; 56(1):45-50. PMC: 2636062. DOI: 10.4103/0301-4738.37595. View

Li W, Zhang X, Lu X, You L, Song Y, Luo Z . 5-Hydroxymethylcytosine signatures in circulating cell-free DNA as diagnostic biomarkers for human cancers. Cell Res. 2017; 27(10):1243-1257. PMC: 5630683. DOI: 10.1038/cr.2017.121. View

Liang J, Yang F, Zhao L, Bi C, Cai B . Physiological and pathological implications of 5-hydroxymethylcytosine in diseases. Oncotarget. 2016; 7(30):48813-48831. PMC: 5217052. DOI: 10.18632/oncotarget.9281. View

Ryu K, Maehr R, Gilchrist C, Long M, Bouley D, Mueller B . The mouse polyubiquitin gene UbC is essential for fetal liver development, cell-cycle progression and stress tolerance. EMBO J. 2007; 26(11):2693-706. PMC: 1888680. DOI: 10.1038/sj.emboj.7601722. View

Li W, Liu M . Distribution of 5-hydroxymethylcytosine in different human tissues. J Nucleic Acids. 2011; 2011:870726. PMC: 3136188. DOI: 10.4061/2011/870726. View

. 11. Microvascular Complications and Foot Care: . Diabetes Care. 2018; 42(Suppl 1):S124-S138. DOI: 10.2337/dc19-S011. View

Lontchi-Yimagou E, Sobngwi E, Matsha T, Kengne A . Diabetes mellitus and inflammation. Curr Diab Rep. 2013; 13(3):435-44. DOI: 10.1007/s11892-013-0375-y. View

Riddy D, Delerive P, Summers R, Sexton P, Langmead C . G Protein-Coupled Receptors Targeting Insulin Resistance, Obesity, and Type 2 Diabetes Mellitus. Pharmacol Rev. 2017; 70(1):39-67. DOI: 10.1124/pr.117.014373. View

Litwak L, Goh S, Hussein Z, Malek R, Prusty V, Khamseh M . Prevalence of diabetes complications in people with type 2 diabetes mellitus and its association with baseline characteristics in the multinational A1chieve study. Diabetol Metab Syndr. 2013; 5(1):57. PMC: 3854020. DOI: 10.1186/1758-5996-5-57. View

10.

Gravina S, Sedivy J, Vijg J . The dark side of circulating nucleic acids. Aging Cell. 2016; 15(3):398-9. PMC: 4854914. DOI: 10.1111/acel.12454. View

11.

Tahiliani M, Koh K, Shen Y, Pastor W, Bandukwala H, Brudno Y . Conversion of 5-methylcytosine to 5-hydroxymethylcytosine in mammalian DNA by MLL partner TET1. Science. 2009; 324(5929):930-5. PMC: 2715015. DOI: 10.1126/science.1170116. View

12.

Ouyang W, Rutz S, Crellin N, Valdez P, Hymowitz S . Regulation and functions of the IL-10 family of cytokines in inflammation and disease. Annu Rev Immunol. 2010; 29:71-109. DOI: 10.1146/annurev-immunol-031210-101312. View

13.

Cubbon R, Kearney M, Wheatcroft S . Endothelial IGF-1 Receptor Signalling in Diabetes and Insulin Resistance. Trends Endocrinol Metab. 2015; 27(2):96-104. DOI: 10.1016/j.tem.2015.11.009. View

14.

Fabregat A, Jupe S, Matthews L, Sidiropoulos K, Gillespie M, Garapati P . The Reactome Pathway Knowledgebase. Nucleic Acids Res. 2017; 46(D1):D649-D655. PMC: 5753187. DOI: 10.1093/nar/gkx1132. View

15.

Hai Z, Zuo W . Aberrant DNA methylation in the pathogenesis of atherosclerosis. Clin Chim Acta. 2016; 456:69-74. DOI: 10.1016/j.cca.2016.02.026. View

16.

Cai J, Chen L, Zhang Z, Zhang X, Lu X, Liu W . Genome-wide mapping of 5-hydroxymethylcytosines in circulating cell-free DNA as a non-invasive approach for early detection of hepatocellular carcinoma. Gut. 2019; 68(12):2195-2205. PMC: 6872444. DOI: 10.1136/gutjnl-2019-318882. View

17.

Zheng Y, Ley S, Hu F . Global aetiology and epidemiology of type 2 diabetes mellitus and its complications. Nat Rev Endocrinol. 2017; 14(2):88-98. DOI: 10.1038/nrendo.2017.151. View

18.

Kriaucionis S, Heintz N . The nuclear DNA base 5-hydroxymethylcytosine is present in Purkinje neurons and the brain. Science. 2009; 324(5929):929-30. PMC: 3263819. DOI: 10.1126/science.1169786. View

19.

Wimmer R, Leopoldi A, Aichinger M, Wick N, Hantusch B, Novatchkova M . Human blood vessel organoids as a model of diabetic vasculopathy. Nature. 2019; 565(7740):505-510. PMC: 7116578. DOI: 10.1038/s41586-018-0858-8. View

20.

Fu Y, He C . Nucleic acid modifications with epigenetic significance. Curr Opin Chem Biol. 2012; 16(5-6):516-24. PMC: 3518615. DOI: 10.1016/j.cbpa.2012.10.002. View