Circulating Levels of Hypoxia-regulating MicroRNAs in Systemic Lupus Erythematosus Patients with Hemolytic Anemia

Abstract

Objective

MicroRNAs are fine regulators for gene expression during the post-transcriptional stage in many autoimmune diseases. HypoxamiRs (miR-210 and miR-21) play an important role in hypoxia and in inflammation-associated hypoxia. Systemic lupus erythematosus (SLE) is a chronic systemic autoimmune disease that would potentiate many pathological complications, including hemolytic anemia. This study aimed to investigate the role of hypoxamiRs in SLE/hemolytic anemia patients.

Methods

This work was designed to analyze the circulating levels of↱ the miR-210 and miR-21 expressions and hypoxia-inducible factor-1α (HIF-α) in SLE/hemolytic anemia patients. SLE activity was evaluated for all patients by SLE Disease Activity Index (SLEDAI). Clinical manifestations/complications and serological/hematological investigations were reported. HIF-α concentration was assayed by ELISA and expression of miR-21 and miR-210 was analyzed by qRT-PCR.

Results

The results indicated that the fold change of the miR-210/miR-21 expressions in plasma was significantly elevated in SLE/hemolytic anemia patients. A strong positive correlation between the miR-210 and miR-21 expression levels was also recorded. Among the associated-disease complications, hypertension, arthritis, oral ulcers, and serositis were associated with a high circulating miR-210 expression, while the occurrence of renal disorders was associated with the increased miR-21 expression. Furthermore, the HIF-α level was remarkably elevated in SLE/hemolytic anemia patients. A high positive correlation was recorded between the HIF-α concentration and miR-210/miR-21 expression levels. The occurrence of oral ulcers, arthritis, and hypertension was associated with the increased HIF-α concentration. On the other hand, SLEDAI and white blood cell count were positively correlated with miR-21/ miR-210. The erythrocyte sedimentation rate was positively correlated with miR-21.

Conclusion

The dysregulation of the circulating miR-210/miR-210/HIF-1α levels in SLE/hemolytic anemia patients advocated that the hypoxia pathway might have an essential role in the pathogenesis and complications of these diseases.

Keywords

circulating hypoxamiRs / miR-210 and miR-21 / hypoxia-inducing factor-1α / hemolytic anemia / systemic lupus erythematosus

References

[1] LeX, YuX, ShenN. Novel insights of microRNAs in the development of systemic lupus erythematosus. Curr Opin Rheumatol, 2017, 29(5): 450-457

[2] BanchereauR, HongS, CantarelB, et al.. Personalized Immunomonitoring Uncovers Molecular Networks that Stratify Lupus Patients. Cell, 2016, 165(3): 551-565

[3] Pacheco-LugoL, Sáenz-GarcíaJ, Navarro QuirozE, et al.. Plasma cytokines as potential biomarkers of kidney damage in patients with systemic lupus erythematosus. Lupus, 2019, 28(1): 34-43

[4] BartelDP. MicroRNAs: genomics, biogenesis, mechanism, and function. Cell, 2004, 116(2): 281-297

[5] AmbrosV. The functions of animal microRNAs. Nature, 2004, 431(7006): 350-355

[6] TaganovKD, BoldinMP, BaltimoreD. MicroRNAs and immunity: tiny players in a big field. Immunity, 2007, 26(2): 133-137

[7] MehtaA, BaltimoreD. MicroRNAs as regulatory elements in immune system logic. Nat Rev Immunol, 2016, 16(5): 279-294

[8] LiuJ, QianC, CaoX. Post-Translational Modification Control of Innate Immunity. Immunity, 2016, 45(1): 15-30

[9] O’ConnellRM, RaoDS, BaltimoreD. microRNA regulation of inflammatory responses. Annu Rev Immunol, 2012, 30: 295-312

[10] BrackenCP, ScottHS, GoodallGJ. A network-biology perspective of microRNA function and dysfunction in cancer. Nat Rev Genet, 2016, 17(12): 719-732

[11] LeungAK, SharpPA. MicroRNA functions in stress responses. Mol Cell, 2010, 40(2): 205-215

[12] CampsC, BuffaFM, ColellaS, et al.. hsa-miR-210 is induced by hypoxia and is an independent prognostic factor in breast cancer. Clin Cancer Res, 2008, 14(5): 1340-1348

[13] GiannakakisA, SandaltzopoulosR, GreshockJ, et al.. miR-210 links hypoxia with cell cycle regulation and is deleted in human epithelial ovarian cancer. Cancer Biol Ther, 2008, 7(2): 255-264

[14] ChanYC, BanerjeeJ, ChoiSY, et al.. miR-210: the master hypoxamir. Microcirculation, 2012, 19(3): 215-223

[15] HuangX, LeQT, GiacciaAJ. MiR-210—micromanager of the hypoxia pathway. Trends Mol Med, 2010, 16(5): 230-237

[16] EltzschigHK, CarmelietP. Hypoxia and inflammation. N Engl J Med, 2011, 364(7): 656-665

[17] WangH, FlachH, OnizawaM, et al.. Negative regulation of Hif1a expression and TH17 differentiation by the hypoxia-regulated microRNA miR-210. Nat Immunol, 2014, 15(4): 393-401

[18] DangEV, BarbiJ, YangHY, et al.. Control of T(H)17/T(reg) balance by hypoxia-inducible factor 1. Cell, 2011, 146(5): 772-784

[19] Crowther M, Podolak-Dawidziak M. Hemolytic Anemia: General Considerations. McMaster Textbook of Internal Medicine. Kraków: Medycyna Praktyczna. 2020

[20] TanEM, CohenAS, FriesJF, et al.. The 1982 revised criteria for the classification of systemic lupus erythematosus. Arthritis Rheum, 1982, 25(11): 1271-1277

[21] BombardierC, GladmanDD, UrowitzMB, et al.. Derivation of the SLEDAI. A disease activity index for lupus patients. The Committee on Prognosis Studies in SLE. Arthritis Rheum, 1992, 35(6): 630-640

[22] ChunHY, ChungJW, KimHA, et al.. Cytokine IL-6 and IL-10 as biomarkers in systemic lupus erythematosus. J Clin Immunol, 2007, 27(5): 461-466

[23] MokMY, WuHJ, LoY, et al.. The relation of interleukin 17 (IL-17) and IL-23 to Th1/Th2 cytokines and disease activity in systemic lupus erythematosus. J Rheumatol, 2010, 37(10): 2046-2052

[24] World Medical Association. World Medical Association Declaration of Helsinki: ethical principles for medical research involving human subjects. JAMA, 2013, 310(20): 2191-2194

[25] LivakKJ, SchmittgenTD. Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method. Methods, 2001, 25(4): 402-408

[26] WuYH, ChanYF, HsiehHL, et al.. Upregulation of miR-210-5p in Systemic Lupus Erythematosus Impairs Silent Clearance of Dead Cell Remnants. FASEB J, 2020, 34(S1): 1-1Supplement: Experimental Biology, Meeting Abstracts

[27] HuangQ, ChenSS, LiJ, et al.. miR-210 expression in PBMCs from patients with systemic lupus erythematosus and rheumatoid arthritis. Ir J Med Sci, 2018, 187(1): 243-249

[28] DaiY, HuangYS, TangM, et al.. Microarray analysis of microRNA expression in peripheral blood cells of systemic lupus erythematosus patients. Lupus, 2007, 16(12): 939-946

[29] HigginsDF, KimuraK, BernhardtWM, et al.. Hypoxia promotes fibrogenesis in vivo via HIF-1 stimulation of epithelial-to-mesenchymal transition. J Clin Invest, 2007, 117(12): 3810-3820

[30] KimuraK, IwanoM, HigginsDF, et al.. Stable expression of HIF-1alpha in tubular epithelial cells promotes interstitial fibrosis. Am J Physiol Renal Physiol, 2008, 295(4): F1023-1029

[31] MaC, WeiJ, ZhanF, et al.. Urinary hypoxia-inducible factor-1alpha levels are associated with histologic chronicity changes and renal function in patients with lupus nephritis. Yonsei Med J, 2012, 53(3): 587-592

[32] DengW, RenY, FengX, et al.. Hypoxia inducible factor-1 alpha promotes mesangial cell proliferation in lupus nephritis. Am J Nephrol, 2014, 40(6): 507-515

[33] GarchowB, Maque AcostaY, et al.. HIF-1α and miR-210 differential and lineage-specific expression in systemic lupus erythematosus. Mol Immunol, 2021, 133: 128-134

[34] RigóJ, MolvarecA, NagyB, et al.. Expression analysis of circulating exosomal hsa-miR-210 in hypertensive disorders of pregnancy: Biomarkers, prediction of preeclampsia. Pregnancy Hypertens, 2016, 6(3): 183