Circulating miRNAs are increasingly suggested as clinical biomarker for diseases. We evaluated the expression of circulating cardiomyocyte-enriched miR-1 and miR-133 by real-time PCR in blood from patients with type 2 diabetes (T2D) without and with coronary artery disease (CAD) and healthy controls, investigated their association with the risk of CAD risk and their potential as biomarkers. The two miRNAs were upregulated in patients with T2D and CAD compared with controls, associated with CAD risk and remained significant after adjustment for multiple confounders. LDL-C was a positive predictor for miR-1 and miR-133, and mean blood pressure was also a positive predictor for miR-133. Both miRNAs strongly distinguished CAD from controls. miR-1 significantly distinguished CAD from T2D with higher diagnostic ability than miR-133, whereas the miR-1/miR-133 combination improved the diagnostic value. Upregulation of circulating miR-1 and miR-133 associate with the risk of CAD in T2D patients and may serve as diagnostic biomarkers.
This is a preview of subscription content, log in to check access.
Buy single article
Instant access to the full article PDF.
Price includes VAT for USA
Subscribe to journal
Immediate online access to all issues from 2019. Subscription will auto renew annually.
This is the net price. Taxes to be calculated in checkout.
Low-density lipoprotein cholesterol
Wild, S., Roglic, G., Green, A., Sicree, R., & King, H. (2004). Global prevalence of diabetes: Estimates for the year 2000 and projections for 2030. Diabetes Care, 27, 1047–1053.
IDF Diabetes Atlas 5th edition. (2011). https://www.idf.org/e-library/epidemiology-research/diabetes-atlas/20-atlas-5th-edition.html.
Alhyas, L., McKay, A., & Majeed, A. (2012). Prevalence of type 2 diabetes in the states of the co-operation council for the Arab states of the Gulf: A systematic review. PLoS One, 7, e40948.
Nathan, D. M. (1993). Long-term complications of diabetes mellitus. The New England Journal of Medicine, 328, 1676–1685.
Laakso, M. (2010). Cardiovascular disease in type 2 diabetes from population to man to mechanisms: The Kelly West Award Lecture 2008. Diabetes Care, 33, 442–449.
Fuster, V., Badimon, L., Badimon, J. J., & Chesebro, J. H. (1992). The pathogenesis of coronary artery disease and the acute coronary syndromes. The New England Journal of Medicine, 326, 242–250.
Libby, P., & Theroux, P. (2005). Pathophysiology of coronary artery disease. Circulation, 111, 3481–3488.
Gillies, C. L., Abrams, K. R., Lambert, P. C., Cooper, N. J., Sutton, A. J., Hsu, R. T., et al. (2007). Pharmacological and lifestyle interventions to prevent or delay type 2 diabetes in people with impaired glucose tolerance: Systematic review and meta-analysis. BMJ, 334(7588), 299.
Li, G., Zhang, P., Wang, J., Gregg, E. W., Yang, W., Gong, Q., et al. (2008). The long-term effect of lifestyle interventions to prevent diabetes in the China Da Qing Diabetes Prevention Study: A 20-year follow-up study. Lancet, 371, 1783–1789.
Bartel, D. P. (2004). Micrornas: Genomics, biogenesis, mechanism, and function. Cell, 116(2), 181–197.
He, L., & Hannon, G. J. (2004). Micrornas: Small rnas with a big role in gene regulation. Nature Reviews. Genetics, 5, 522–531.
Kloosterman, W. P., & Plasterk, R. H. (2006). The diverse functions of microRNAs in animal development and disease. Developmental Cell, 11, 441–450.
Latronico, M. V., Catalucci, D., & Condorelli, G. (2007). Emerging role of microRNAs in cardiovascular biology. Circulation Research, 101, 1225–1236.
Pandey, A. K., Agarwal, P., Kaur, K., & Datta, M. (2009). MicroRNAs in diabetes: Tiny players in big disease. Cellular Physiology and Biochemistry, 23, 221–232.
Ardekani, A. M., & Naeini, M. (2010). The role of microRNAs in human diseases. Avicenna Journal of Medical Biotechnology, 2, 161–179.
Condorelli, G., Latronico, M. V., & Cavarretta, E. (2014). microRNAs in cardiovascular diseases current: Knowledge and the road ahead. Journal of the American College of Cardiology, 63(21), 2177–2187.
Mitchell, P. S., Parkin, R. K., Kroh, E. M., Fritz, B. R., Wyman, S. K., Pogosova-Agadjanyan, E. L., et al. (2008). Circulating microRNAs as stable blood-based markers for cancer detection. Proceedings of the National Academy of Sciences of the United States of America, 105, 10513–10518.
Chen, X., Ba, Y., Ma, L., Cai, X., Yin, Y., Wang, K., et al. (2008). Characterization of microRNAs in serum: A novel class of biomarkers for diagnosis of cancer and other diseases. Cell Research, 18, 997–1006.
Meder, B., Keller, A., Vogel, B., Haas, J., Sedaghat-Hameddani, F., Kayvanpour, E., et al. (2011). MicroRNA signatures in total peripheral blood as novel biomarkers for acute myocardial infarction. Basic Research in Cardiology, 106, 13–23.
Al-Kafaji, G., Al-Mahroos, G., Alsayed, N. A., Hasan, Z. A., Nawaz, S., & Bakhiet, M. (2015). Peripheral blood microRNA-15a as a potential biomarker for type 2 diabetes mellitus and pre-diabetes. Molecular Medicine Reports, 12(5), 7485–7490.
Al-Kafaji, G., Al-Mahroos, G., Al-Muhtaresh, H. A., Skrypnyk, C., Sabry, M. A., & Ramadan, A. R. (2016). Decreased expression of circulating microRNA-126 in patients with type 2 diabetic nephropathy: A potential blood-based biomarker. Experimental and Therapeutic Medicine, 12(2), 815–822.
Al-Kafaji, G., Al Naieb, Z. T., & Bakhiet, M. (2016). Increased oncogenic microRNA-18a expression in peripheral blood of patients with prostate cancer: A potential role as new noninvasive biomarker. Oncology Letters, 11(2), 1201–1120.
Al-Muhtaresh, H., & Al-Kafaji, G. (2018). Evaluation of two-diabetes related microRNAs suitability as earlier blood biomarkers for detecting prediabetes and type 2 diabetes mellitus. Journal of Clinical Medical, 7(2), 12.
Fichtlscherer, S., De Rosa, S., Fox, H., Schwietz, T., Fischer, A., Liebetrau, C., et al. (2010). Circulating microRNAs in patients with coronary artery disease. Circulation Research, 107, 677–684.
Rao, P. K., Kumar, R. M., Farkhondeh, M., Baskerville, S., & Lodish, H. F. (2006). Myogenic factors that regulate expression of muscle-specific microRNAs. Proceedings of the National Academy of Sciences of the United States of America, 103, 8721–8726.
Townley-Tilson, W. H., Callis, T. E., & Wang, D. (2010). MicroRNAs 1, 133, and 206: Critical factors of skeletal and cardiac muscle development, function, and disease. The International Journal of Biochemistry & Cell Biology, 42, 1252–1255.
Chistiakov, D. A., Orekhov, A. N., & Bobryshev, Y. V. (2016). Cardiac-specific miRNA in cardiogenesis, heart function, and cardiac pathology (with focus on myocardial infarction). Journal of Molecular and Cellular Cardiology, 94, 107–121.
Zhao, Y., Samal, E., & Srivastava, D. (2005). Serum response factor regulates a muscle-specific microRNA that targets Hand2 during cardiogenesis. Nature, 436, 214–220.
Ikeda, S., He, A., Kong, S. W., Lu, J., Bejar, R., Bodyak, N., et al. (2009). MicroRNA-1 negatively regulates expression of the hypertrophy-associated calmodulin and Mef2a genes. Molecular and Cellular Biology, 29, 2193–2204.
Chen, J. F., Mandel, E. M., Thomson, J. M., Wu, Q., Callis, T. E., Hammond, S. M., et al. (2006). The role of microRNA-1 and microRNA-133 in skeletal muscle proliferation and differentiation. Nature Genetics, 38, 228–233.
Wang, G., Zhu, J., Zhang, J., Li, Q., Li, Y., He, J., et al. (2010). Circulating microRNA: A novel potential biomarker for early diagnosis of acute myocardial infarction in humans. European Heart Journal, 31, 659–666.
Kuwabara, Y., Ono, K., Horie, T., Nishi, H., Nagoa, K., Kinoshita, M., et al. (2011). Increased microRNA-1 and microRNA-133a levels in serum of patients with cardiovascular disease indicate myocardial damage. Circulation. Cardiovascular Genetics, 4(4), 446–454.
Briasoulis, A., Tousoulis, D., Vogiatzi, G., Siasos, G., Papageorgiou, N., Oikonomou, E., et al. (2013). MicroRNAs: Biomarkers for cardiovascular disease in patients with diabetes mellitus. Current Topics in Medicinal Chemistry, 13(13), 1533–1539.
Al-Kafaji, G., Al-Mahroos, G., Al-Muhtaresh, H., Sabry, M. A., Abdul Razzak, R., & Salem, A. H. (2017). Circulating endothelium-enriched microRNA-126 as a potential biomarker for coronary artery disease in type 2 diabetes mellitus patients. Biomarkers, 22(3–4), 268–278.
AL-Subaihi, A. (2003). Sample size determination. Influencing factors and calculation strategies for survey research. Saudi Medical Journal, 24(4), 323–330.
Alberti, K. G., & Zimmet, P. Z. (1998). Definition, diagnosis and classification of diabetes mellitus and its complications. Part 1: Diagnosis and classification of diabetes mellitus provisional report of a WHO consultation. Diabetic Medicine, 15, 539–553.
Wang, X. (2009). A PCR-based platform for microRNA expression profiling studies. RNA, 15, 716–723.
Feng, Y., Niu, L.-L., Wei, W., Zhang, W.-Y., Li, X.-Y., Cao, J.-H., & Zhao, S.-H. (2013). A feedback circuit between miR-133 and the ERK1/2 pathway involving an exquisite mechanism for regulating myoblast proliferation and differentiation. Cell Death & Disease, 4, e934.
Wong, L., Lee, K., Russell, I., & Chen, C. (2007). Endogenous controls for real time quantitation of miRNA using TaqManVR MicroRNA assays. New York: Macmillan Publishers Ltd..
Roggli, E., Britan, A., Gattesco, S., Lin-Marq, N., Abderrahmani, A., Meda, P., et al. (2010). Involvement of microRNAs in the cytotoxic effects exerted by proinflammatory cytokines on pancreatic beta-cells. Diabetes, 59, 978–986.
Poy, M. N., Hausser, J., Trajkovski, M., Braunc, M., Collinsc, S., Rorsmanc, P., et al. (2009). miR-375 maintains normal pancreatic alpha- and beta-cell mass. Proceedings of the National Academy of Sciences of the United States of America, 106, 5813–5818.
Care, A., Catalucci, D., Felicetti, F., Bonci, D., Addario, A., Gallo, P., et al. (2007). MicroRNA-133 controls cardiac hypertrophy. Nature Medicine, 13, 613–618.
Terentyev, D., Belevych, A. E., Terentyeva, R., Martin, M. M., Malana, G. E., Kuhn, D. E., et al. (2009). miR-1 overexpression enhances Ca(2+) release and promotes cardiac arrhythmogenesis by targeting PP2A regulatory subunit B56alpha and causing CaMKII-dependent hyperphosphorylation of RyR2. Circulation Research, 104, 514–521.
Luo, X., Lin, H., Pan, Z., Xiao, J., Zhang, Y., Lu, Y., et al. (2008). Down-regulation of miR-1/miR-133 contributes to re-expression of pacemaker channel genes HCN2 and HCN4 in hypertrophic heart. The Journal of Biological Chemistry, 283, 20045–20052.
Williams, A. H., Liu, N., van Rooij, E., & Olson, E. N. (2009). MicroRNA control of muscle development and disease. Current Opinion in Cell Biology, 21, 461–469.
Liu, N., Bezprozvannaya, S., Williams, A. H., Qi, X., Richardson, J. A., Bassel-Duby, R., et al. (2008). microRNA-133a regulates cardiomyocyte proliferation and suppresses smooth muscle gene expression in the heart. Genes & Development, 22, 3242–3254.
D’Alessandra, Y., Devanna, P., Limana, F., Straino, S., Di Carlo, A., Brambilla, P. G., et al. (2010). Circulating microRNAs are new and sensitive biomarkers of myocardial infarction. European Heart Journal, 31, 2765–2773.
Matheus, A. S., Tannus, L. R., Cobas, R. A., Palm, A. C. C., Negrato, C. A., & Gomes, M. B. (2013). Impact of diabetes on cardiovascular disease: An update. International Journal of Hypertension, 2013, 653789.
Dokken, B. B. (2008). The pathophysiology of cardiovascular disease and diabetes: Beyond blood pressure and lipids. Diabetes Spectrum: A Publication of the American Diabetes Association, 21(3), 160–165.
Grundy, S. M., Pasternak, R., Greenland, P., Smith, S., Jr., & Fuster, V. (1999). Assessment of cardiovascular risk by use of multiple-risk-factor assessment equations. A statement for healthcare professionals from the American heart association and the American college of cardiology. Circulation, 100, 1481–1492.
Howard, B. V., Robbins, D. C., Sievers, M. L., et al. (2000). LDL cholesterol as a strong predictor of coronary heart disease in diabetic individuals with insulin resistance and low LDL. The Strong heart study. Arteriosclerosis, Thrombosis, and Vascular Biology, 20, 830–835.
Escobar, E. (2002). Hypertension and coronary heart disease. Journal of Human Hypertension, 16(1), S61–S63.
Otsuka, T., Takada, H., Nishiyama, Y., Kodani, E., Saiki, Y., Kato, K., & Kawada, T. (2016). Dyslipidemia and the risk of developing hypertension in a working-age male population. Journal of the American Heart Association, 5, e003053.
de Gonzalo-Calvo, D., van der Meer, R. W., Rijzewijk, L. J., Smit, J. W. A., Revuelta-Lopez, E., Nasarre, L., et al. (2017). Serum microRNA-1 and microRNA-133a levels reflect myocardial steatosis in uncomplicated type 2 diabetes. Scientific Reports, 7(1), 47.
Zhang, Z., Joyce, B. T., Kresovich, J. K., Zheng, Y., Zhong, J., Patel, R., et al. (2017). Blood pressure and expression of microRNAs in whole blood. PLoS One, 12(3), e0173550.
Jepsen, A. M., Langsted, A., Varbo, A., Bang, L. E., Kamstrup, P. R., & Nordestgaard, B. G. (2016). Increased remnant cholesterol explains part of residual risk of all-cause mortality in 5414 patients with ischemic heart disease. Clinical Chemistry, 62(4), 593–604.
Sampson, U. K., Fazio, S., & Linton, M. F. (2012). Residual cardiovascular risk despite optimal LDL cholesterol reduction with statins: The evidence, etiology, and therapeutic challenges. Current Atherosclerosis Reports, 14(1), 1–10.
Fruchart, J. C., Davignon, J., Hermans, M. P., Al-Rubeaan, K., Amarenco, P., Assmann, G., Barter, P., Betteridge, J., Bruckert, E., Cuevas, A., Farnier, M., et al. (2014). Residual macrovascular risk in 2013: What have we learned? Cardiovascular Diabetology, 13(1), 26.
Cui, Y., Blumenthal, R. S., Flaws, J. A., Whiteman, M. K., Langenberg, P., Bachorik, P. S., & Bush, T. L. (2001). Non-high-density lipoprotein cholesterol level as a predictor of cardiovascular disease mortality. Archives of Internal Medicine, 161, 1413–1419.
Lu, W., Resnick, H. E., Jablonski, K. A., Jones, K. L., Jain, A. K., Howard, W. J., Robbins, D. C., & Howard, B. V. (2003). Non-HDL cholesterol as a predictor of cardiovascular disease in type 2 diabetes. The strong heart study. Diabetes Care, 26(1), 16–23.
Kroh, E. M., Parkin, R. K., Mitchell, P. S., & Tewari, M. (2010). Analysis of circulating microRNA biomarkers in plasma and serum using quantitative reverse transcription-PCR (qRT-PCR). Methods, 50, 298–301.
Li, Y., & Kowdley, K. V. (2012). Method for microRNA isolation from clinical serum samples. Analytical Biochemistry, 431(1), 69–75.
We would like to acknowledge the technical support of the Research Unit staff at Al-Jawhara Centre for Molecular Medicine, Genetics and Inherited Disorders in the College of Medicine and Medical Sciences, Arabian Gulf University.
The present study was supported by a research grant (No. 81) by the College of Medicine and Medical Sciences, Arabian Gulf University, Kingdom of Bahrain and a PhD Research Grant by Kingdom of Saudi Arabia.
Conflict of Interest
The authors declare that they have no conflict of interest.
Ethical approval to conduct the current study was obtained from the Medical Research and Ethics Committee in the College of Medicine and Medical Sciences, Arabian Gulf University, Kingdom of Bahrain. The participants were given a complete description of the study, and provided written informed consents according to the guidelines of the College of Medicine and Medical Sciences, Arabian Gulf University, Kingdom of Bahrain.
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Clinical Relevance of the Study
• Currently, there are no biomarkers for early detection of type 2 diabetes-associated macrovascular complications.
• Circulating miRNAs are increasingly suggested as clinical biomarker for several diseases.
• We show that the upregulation of circulating cardiomyocyte-enriched miR-1 and miR-133 are associated with the risk of coronary artery disease in type 2 diabetes patients and could serve as diagnostic biomarkers.
Associate Editor Craig Stolen oversaw the review of this article
About this article
Cite this article
Al-Muhtaresh, H.A., Salem, A.H. & Al-Kafaji, G. Upregulation of Circulating Cardiomyocyte-Enriched miR-1 and miR-133 Associate with the Risk of Coronary Artery Disease in Type 2 Diabetes Patients and Serve as Potential Biomarkers. J. of Cardiovasc. Trans. Res. 12, 347–357 (2019). https://doi.org/10.1007/s12265-018-9857-2
- Circulating miRNAs
- Type 2 diabetes
- Coronary artery disease