Plasma-derived exosomes implement miR-126-associated regulation of cytokines secretion in PBMCs of CHF patients in vitro

Larysa Natrus; Dmytro Labudzynskyi; Petro Muzychenko; Petro Chernovol; Yuliia Klys

doi:10.23750/abm.v93i3.12449

Plasma-derived exosomes implement miR-126-associated regulation of cytokines secretion in PBMCs of CHF patients in vitro

Authors

Larysa V. Natrus Bogomolets National Medical University, Kyiv, Ukraine https://orcid.org/0000-0003-1763-0618
Dmytro O. Labudzynskyi Palladin Institute of Biochemistry of the National Academy of Sciences of Ukraine, Kyiv, Ukraine https://orcid.org/0000-0003-4389-6049
Petro F. Muzychenko Bogomolets National Medical University, Kyiv, Ukraine
Petro A. Chernovol Bogomolets National Medical University, Kyiv, Ukraine
Yuliia G. Klys Bogomolets National Medical University, Kyiv, Ukraine

Keywords:

Plasma-derived exosomes, PBMC cells, congestive heart failure, miRNA-126, cytokines, paracrine secretion

Abstract

Background The investigation of regulatory effects of intra-exosomal compounds, especially microRNAs, has promising therapeutic prospects in the treatment of numerous diseases, including cardiovascular disorders. In this study, we investigated the effect of healthy donors` plasma exosomes (HDPE) on the production of cytokines by PBMC cells of patients with congestive heart failure (CHF) and showed the integral role of miRNA-126 in CHF-mediated changes of mononuclear paracrine secretion. Methods Peripheral blood mononuclear cells (PBMСs) were isolated from a peripheral blood of fifteen patients with CHF (age, 66,8±9,8 years; left ventricular ejection fraction, 44±19%). The concentration of cytokines (IL-10, ICAM-1, VEGF-A, TNF-α and MCP-1) in culture medium and PBMCs was measured by ELISA. The level of miRNA-126 expression in PBMCs was performed by real-time PCR. Results Dramatic increase of ICAM-1 level in activated PBMCs of CHF patients, as well as an increase of the IL-10, ICAM-1 and TNF-α levels in the culture medium was observed. It was accompanied by CHF-related miRNA-126 overexpression in PBMCs. HDPE treatment distinguished by a tendency to reduction in miRNA-126 expression by CHF PBMCs and correlated with upregulation of IL-10, ICAM-1, TNF-α and MCP-1 with normalization of cytokines secretion. Conclusions The altered paracrine secretion of cytokines by CHF PBMCs and miRNA-126 overexpression in vitro was found. HDPE treatment modulated production and secretion of most of studied cytokines by CHF PBMCs in vitro. The experimental application of exosomes for the modulation of paracrine secretion and PBMCs cellular functions may be promising for CVD therapy, including endothelial dysfunction and CHF.

Author Biographies

Larysa V. Natrus, Bogomolets National Medical University, Kyiv, Ukraine

Prof., MD, Head of Department of Modern Technologies of Medical Diagnostics and Treatment; Bogomolets National Medical University, Kyiv, Ukraine

Dmytro O. Labudzynskyi, Palladin Institute of Biochemistry of the National Academy of Sciences of Ukraine, Kyiv, Ukraine

Research Fellow (PhD) in the Department of Biochemistry Vitamins and Coenzymes; Palladin Institute of Biochemistry of the National Academy of Sciences of Ukraine, Kyiv, Ukraine

Petro F. Muzychenko, Bogomolets National Medical University, Kyiv, Ukraine

Professor, MD, Department of Operative Surgery and Topographic Anatomy; Bogomolets National Medical University, Kyiv, Ukraine

Petro A. Chernovol, Bogomolets National Medical University, Kyiv, Ukraine

Laboratory assistant in the Department of Modern Technologies of Medical Diagnostics and Treatment; Bogomolets National Medical University, Kyiv, Ukraine

Yuliia G. Klys, Bogomolets National Medical University, Kyiv, Ukraine

PhD, Head of the Laboratory of Experimental Research; Research Institute of Experimental and Clinical Medicine; Bogomolets National Medical University, Kyiv, Ukraine

References

Hessel FP. Overview of the socio-economic consequences of heart failure. Cardiovasc Diagn Ther. 2021; 11(1):254-262.

Metra M, Teerlink JR. Heart failure. Lancet 2017; 390 (10106):1981–1995.

Boorsma EM, Ter Maaten JM, Damman K, Dinh W, Gustafsson F, Goldsmith S, Burkhoff D, Zannad F, Udelson JE, Voors AA. Congestion in heart failure: a contemporary look at physiology, diagnosis and treatment. Nat Rev Cardiol. 2020; 17(10):641-655.

Alem MM. Endothelial Dysfunction in Chronic Heart Failure: Assessment, Findings, Significance, and Potential Therapeutic Targets. Int J Mol Sci. 2019; 20(13):3198.

Swirski FK, Nahrendorf M. Cardioimmunology: the immune system in cardiac homeostasis and disease. Nat Rev Immunol. 2018; 18(12):733-744.

Riehle C, Bauersachs J. Key inflammatory mechanisms underlying heart failure. Entzündungsmechanismen bei Herzinsuffizienz. Herz. 2019; 44(2):96-106.

Liu W, Ru L, Su C, Qi S, Qi X. Serum Levels of Inflammatory Cytokines and Expression of BCL2 and BAX mRNA in Peripheral Blood Mononuclear Cells and in Patients with Chronic Heart Failure. Med Sci Monit. 2019; 25:2633-2639.

Shirakawa R, Yokota T, Nakajima T, Takada S, Yamane M, Furihata T, Maekawa S, Nambu H, Katayama T, Fukushima A, Saito A, Ishimori N, Dela F, Kinugawa S, Anzai T. Mitochondrial reactive oxygen species generation in blood cells is associated with disease severity and exercise intolerance in heart failure patients. Sci Rep. 2019; 9(1):14709.

Eskandari V, Amirzargar AA, Mahmoudi MJ, Rahnemoon Z, Rahmani F, Sadati S, Rahmati Z, Gorzin F, Hedayat M, Rezaei N. Gene expression and levels of IL-6 and TNFα in PBMCs correlate with severity and functional class in patients with chronic heart failure. Ir J Med Sci. 2018; 187(2):359-368.

Wong LL, Wang J, Liew OW, Richards AM, Chen YT. MicroRNA and Heart Failure. Int J Mol Sci. 2016; 17(4):502.

Ionescu RF, Cretoiu SM. MicroRNAs as monitoring markers for right-sided heart failure and congestive hepatopathy. J Med Life. 2021; 14(2):142-147.

Wei XJ, Han M, Yang FY, Wei GC, Liang ZG, Yao H, et al. Biological significance of miR-126 expression in a trial fibrillation and heart failure. Braz J Med Biol Res. 2015; 48(11):983–989.

Wang X, Lian Y, Wen X, et al. Expression of miR-126 and its potential function in coronary artery disease. Afr Health Sci. 2017; 17(2):474-480.

Zhong L, Simard MJ, Huot J. Endothelial microRNAs regulating the NF-κB pathway and cell adhesion molecules during inflammation. FASEB J. 2018; 32(8):4070-4084.

Khanaghaei M, Tourkianvalashani F, Hekmatimoghaddam S, et al. Circulating miR-126 and miR-499 reflect progression of cardiovascular disease; correlations with uric acid and ejection fraction. Heart Int. 2016; 11(1):e1-e9.

Mocharla, Pavani; Briand, Sylvie; Giannotti, Giovanna; Dörries, Carola; Jakob, Philipp; Paneni, Francesco; Lüscher, Thomas F; Landmesser, Ulf. AngiomiR-126 expression and secretion from circulating CD34+ and CD14+ PBMCs: role for pro-angiogenic effects and alterations in type-2 diabetics. Blood 2013; 121(1):226-236.

Qin A, Wen Z, Zhou Y, et al. MicroRNA-126 regulates the induction and function of CD4(+) Foxp3(+) regulatory T cells through PI3K/AKT pathway. J Cell Mol Med. 2013; 17(2):252-264.

Capetini VC, Quintanilha BJ, Sampaio GR, Ferreira FM, Rogero M. Blood Orange Juice Intake Modulates the Expression of miR-126–3p and let-7f-5p in PBMC of Overweight and Insulin Resistance Women. Curr Dev Nutr. 2021; 5(Suppl 2):936.

He N, Zhang Y, Zhang S, Wang D, Ye H. Exosomes: Cell-Free Therapy for Cardiovascular Diseases. J Cardiovasc Transl Res. 2020; 13(5):713-721.

Zamani P, Fereydouni N, Butler AE, Navashenaq JG, Sahebkar A. The therapeutic and diagnostic role of exosomes in cardiovascular diseases. Trends Cardiovasc Med. 2019; 29(6):313-323.

Zhou Y, Ming J, Li Y, Li B, Deng M, Ma Y, Chen Z, Zhang Y, Li J, Liu S. Exosomes derived from miR-126-3p-overexpressing synovial fibroblasts suppress chondrocyte inflammation and cartilage degradation in a rat model of osteoarthritis. Cell Death Discov. 2021; 7(1):37.

Antonakos N, Tsaganos T, Oberle V, Tsangaris I, Lada M, Pistiki A, Machairas N, Souli M, Bauer M, Giamarellos-Bourboulis EJ. Decreased cytokine production by mononuclear cells after severe gram-negative infections: early clinical signs and association with final outcome. Crit Care. 2017; 21(1):48.

Xu S, Zhang J, Liu J, Ye J, Xu Y, Wang Z, Yu J, Ye D, Zhao M, Feng Y, Pan W, Wang M, Wan J. The role of interleukin-10 family members in cardiovascular diseases. Int Immunopharmacol. 2021; 94:107475.

Byrne A, Reen DJ. Lipopolysaccharide induces rapid production of IL-10 by monocytes in the presence of apoptotic neutrophils. J Immunol. 2002; 168(4):1968-77.

Suh JH, Joo HS, Hong EB, Lee HJ, Lee JM. Therapeutic Application of Exosomes in Inflammatory Diseases. Int J Mol Sci. 2021; 22(3):1144.

Tang TT, Wang B, Lv LL, Liu BC. Extracellular vesicle-based Nanotherapeutics: Emerging frontiers in anti-inflammatory therapy. Theranostics. 2020; 10(18):8111-8129.

Ichiki T, Jougasaki M, Setoguchi M, Imamura J, Nakashima H, Matsuoka T, Sonoda M, Nakamura K, Minagoe S, Tei C. Cardiotrophin-1 stimulates intercellular adhesion molecule-1 and monocyte chemoattractant protein-1 in human aortic endothelial cells. Am J Physiol Heart Circ Physiol. 2008; 294(2):H750-63.

Byrkjeland R, Nilsson BB, Westheim AS, Arnesen H, Seljeflot I. Inflammatory markers as related to disease severity in patients with chronic heart failure: limited effects of exercise training. Scand J Clin Lab Invest. 2011; 71(7):598-605.

Pearson MJ, Mungovan SF, Smart NA. Effect of aerobic and resistance training on inflammatory markers in heart failure patients: systematic review and meta-analysis. Heart Fail Rev. 2018; 23(2):209-223.

Möst J, Schwaeble W, Drach J, Sommerauer A, Dierich MP. Regulation of the expression of ICAM-1 on human monocytes and monocytic tumor cell lines. J Immunol. 1992; 148(6):1635-42.

Sáez T, de Vos P, Kuipers J, Sobrevia L, Faas MM. Exosomes derived from monocytes and from endothelial cells mediate monocyte and endothelial cell activation under high d-glucose conditions. Immunobiology. 2019; 224(2):325-333.

Morimoto H, Takahashi M. Role of monocyte chemoattractant protein-1 in myocardial infarction. Int J Biomed Sci. 2007; 3(3):159-167.

Georgakis MK, Gill D, Rannikmäe K, et al. Genetically Determined Levels of Circulating Cytokines and Risk of Stroke. Circulation. 2019; 139(2):256-268.

Hohensinner PJ, Kaun C, Rychli K, Ben-Tal Cohen E, Kastl SP, Demyanets S, Pfaffenberger S, Speidl WS, Rega G, Ullrich R, Maurer G, Huber K, Wojta J. Monocyte chemoattractant protein (MCP-1) is expressed in human cardiac cells and is differentially regulated by inflammatory mediators and hypoxia. FEBS Lett. 2006; 580(14):3532-8.

Ruiz Silva M, van der Ende-Metselaar H, Mulder HL, Smit JM, Rodenhuis-Zybert IA. Mechanism and role of MCP-1 upregulation upon chikungunya virus infection in human peripheral blood mononuclear cells. Sci Rep. 2016; 6:32288.

Gu W, Yao L, Li L, et al. ICAM-1 regulates macrophage polarization by suppressing MCP-1 expression via miR-124 upregulation. Oncotarget. 2017; 8(67):111882-111901.

Bui TM, Wiesolek HL, Sumagin R. ICAM-1: A master regulator of cellular responses in inflammation, injury resolution, and tumorigenesis. J Leukoc Biol. 2020; 108(3):787-799.

Nash M, McGrath JP, Cartland SP, Patel S, Kavurma MM. Tumour necrosis factor superfamily members in ischaemic vascular diseases. Cardiovasc Res. 2019; 115(4):713-720.

Satoh M, Iwasaka J, Nakamura M, Akatsu T, Shimoda Y, Hiramori K. Increased expression of tumor necrosis factor-alpha converting enzyme and tumor necrosis factor-alpha in peripheral blood mononuclear cells in patients with advanced congestive heart failure. Eur J Heart Fail. 2004; 6(7):869-75.

Arakawa H, Ikeda U, Hojo Y, et al. Decreased serum vascular endothelial growth factor concentrations in patients with congestive heart failure. Heart. 2003; 89(2):207-208.

Jaipersad AS, Lip GY, Silverman S, Shantsila E. The role of monocytes in angiogenesis and atherosclerosis. J Am Coll Cardiol. 2014; 63(1):1-11.

Apostolakis S, Lip GY, Shantsila E. Monocytes in heart failure: relationship to a deteriorating immune overreaction or a desperate attempt for tissue repair? Cardiovasc Res. 2010; 85(4):649-60.

Wu WK, Llewellyn OP, Bates DO, Nicholson LB, Dick AD. IL-10 regulation of macrophage VEGF production is dependent on macrophage polarisation and hypoxia. Immunobiology. 2010; 215(9-10):796-803.

Schwarzenbach H, Gahan PB. MicroRNA Shuttle from Cell-To-Cell by Exosomes and Its Impact in Cancer. Noncoding RNA. 2019; 5(1):28.

Vicencio JM, Yellon DM, Sivaraman V, Das D, Boi-Doku C, Arjun S, Zheng Y, Riquelme JA, Kearney J, Sharma V, Multhoff G, Hall AR, Davidson SM. Plasma exosomes protect the myocardium from ischemia-reperfusion injury. J Am Coll Cardiol. 2015; 65(15):1525-36.

Pomatto MAC, Bussolati B, D'Antico S, et al. Improved Loading of Plasma-Derived Extracellular Vesicles to Encapsulate Antitumor miRNAs. Mol Ther Methods Clin Dev. 2019; 13:133-144.