Reduced miR-99a-3p levels in systemic lupus erythematosus may promote B cell proliferation via NCAPG and the PI3K/AKT signaling pathway

Abstract

Background

Systemic lupus erythematosus is an immunologically dysregulated disease characterized by the presence of multiple autoantibodies. In SLE, B lymphocytes contribute to the dysregulated production of autoantibodies and cytokines. Recently, we discovered that miR-99a-3p binds to both EIF4EBP1 and NCAPG mRNA and that lowering miR-99a-3p can promote B cell autophagy in SLE by increasing EIF4EBP1 expression. However, the functions of miR-99a-3p and NCAPG in SLE have not been extensively investigated.

Objective

This work aims to evaluate the levels of miR-99a-3p and NCAPG expression in SLE B cells and to determine whether the aberrant expression of miR-99a-3p and NCAPG contributes to the pathological mechanisms in SLE.

Methods

B lymphocytes were obtained through immunomagnetic negative selection. Using RT-qPCR, miR-99a-3p and NCAPG mRNA expressions in B lymphocytes and in the BALL-1 cell line were measured. To determine the relative abundance of NCAPG, PI3K, p-PI3K, AKT, and p-AKT, we normalize them to the level of β-actin using Western blotting. Evaluation of miR-99a-3p and NCAPG’s impact on cell proliferation was done utilizing CCK-8 assay. Using flow cytometry, the cell cycle and apoptosis were both measured.

Results

Comparing SLE B cells to healthy controls, miR-99a-3p expression was significantly downregulated. Additionally, it was observed that SLE B cells had significantly higher NCAPG mRNA expression. Blocking miR-99a-3p expression in BALL-1 cells with an antagomir elevated NCAPG expression, facilitated PI3K/AKT pathway activation, improved cell proliferation, raised the fraction of S-phase cells, and prevented cell apoptosis. The opposite effects of upregulated miR-99a-3p levels on BALL-1 cells were observed by using an agomir. Furthermore, the effect of decreased miR-99a-3p expression on cell proliferation was partially mediated by elevating NCAPG levels and activating the PI3K/AKT pathway.

Conclusion

Our research indicates that lower miR-99a-3p expression in SLE B cells appears to boost B cell number via the NCAPG and PI3K/AKT pathways.

Keywords

miR-99a-3p NCAPG PI3K/AKT pathway proliferation systemic lupus erythematosus

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References

Kiriakidou

Ching

. Systemic Lupus Erythematosus. Ann Intern Med 2020; 172: ITC81–ITC96.

Parks

de Souza Espindola Santos

Barbhaiya

, et al. Understanding the role of environmental factors in the development of systemic lupus erythematosus. Best Pract Res Clin Rheumatol 2017; 31: 306–320.

Yap

DYH

Chan

. B cell abnormalities in systemic lupus erythematosus and lupus nephritis-role in pathogenesis and effect of immunosuppressive treatments. Int J Mol Sci 2019; 20: E6231.

Dhillon

. Telitacicept: first approval. Drugs 2021; 81: 1671–1675.

Shen

. miRNAs in the pathogenesis of systemic lupus erythematosus. Int J Mol Sci 2015; 16: 9557–9572.

Schell

Rahman

ZSM

. miRNA-mediated control of B cell responses in immunity and SLE. Front Immunol 2021; 12: 683710.

Luo

Ding

Liao

, et al. Excessive miR-152-3p results in increased BAFF expression in SLE B-cells by inhibiting the KLF5 expression. Front Immunol 2019; 10: 1127.

Yang

Huang

, et al. Decreased miR-4512 levels in monocytes and macrophages of individuals with systemic lupus erythematosus contribute to innate immune activation and neutrsophil NETosis by targeting TLR4 and CXCL2. Front Immunol 2021; 12: 756825.

Yang

Deng

. Targeting of EIF4EBP1 by miR-99a-3p affects the functions of B lymphocytes via autophagy and aggravates SLE disease progression. J Cell Mol Med 2021; 25: 10291–10305.

10.

Sutani

Sakata

Nakato

, et al. Condensin targets and reduces unwound DNA structures associated with transcription in mitotic chromosome condensation. Nat Commun 2015; 6: 7815.

11.

Gong

Fan

, et al. NCAPG promotes the proliferation of hepatocellular carcinoma through PI3K/AKT signaling. Onco Targets Ther 2019; 12: 8537–8552.

12.

Lin

Pan

, et al. NCAPG promotes the progression of lung adenocarcinoma via the TGF-β signaling pathway. Cancer Cell Int 2021; 21: 443.

13.

Sun

, et al. NCAPG, mediated by miR-378a-3p, regulates cell proliferation, cell cycle progression, and apoptosis of oral squamous cell carcinoma through the GSK-3β/β-catenin signaling. Neoplasma 2021; 68: 1201–1211.

14.

Hochberg

. Updating the American College of Rheumatology revised criteria for the classification of systemic lupus erythematosus. Arthritis Rheum 1997; 40: 1725.

15.

Nie

Zhao

Zhang

. B cell aberrance in lupus: the ringleader and the solution. Clin Rev Allergy Immunol 2022; 62: 301–323.

16.

Gonzalez-Martin

Adams

Lai

, et al. The microRNA miR-148a functions as a critical regulator of B cell tolerance and autoimmunity. Nat Immunol 2016; 17: 433–440.

17.

Wang

Chen

Qiu

, et al. Antagonizing miR-7 suppresses B cell hyperresponsiveness and inhibits lupus development. J Autoimmun 2020; 109: 102440.

18.

Shi

, et al. Downregulated miR-29a promotes B cell overactivation by upregulating Crk-like protein in systemic lupus erythematosus. Mol Med Rep 2020; 22: 841–849.

19.

Guo

Zhang

, et al. Delivery of AntagomiR-7 through polymer nanoparticles for assisting B Cell to alleviate systemic lupus erythematosus. Front Bioeng Biotechnol 2023; 11: 1180302.

20.

Eniafe

Jiang

. MicroRNA-99 family in cancer and immunity, 12. Wiley Interdiscip Rev RNA, 2021, p. e1635.

21.

Zhao

Cheng

, et al. Up-regulation of the expressions of MiR-149-5p and MiR-99a-3p in exosome inhibits the progress of pituitary adenomas. Cell Biol Toxicol 2021; 37: 633–651.

22.

Yan

Zhang

Cen

, et al. Expression of microRNA-99a-3p in prostate cancer based on bioinformatics data and meta-analysis of a literature review of 965 cases. Med Sci Monit Int Med J Exp Clin Res 2018; 24: 4807–4822.

23.

Jaiswal

Maurya

, et al. Lin28B regulates angiotensin II-mediated let-7c/miR-99a MicroRNA formation consequently affecting macrophage polarization and allergic inflammation. Inflammation 2020; 43: 1846–1861.

24.

Warth

Hoefig

Hiekel

, et al. Induced miR-99a expression represses Mtor cooperatively with miR-150 to promote regulatory T-cell differentiation. EMBO J 2015; 34: 1195–1213.

25.

Hua

Wang

, et al. Aberrant expression of microRNA in CD4+ cells contributes to Th17/Treg imbalance in primary immune thrombocytopenia. Thromb Res 2019; 177: 70–78.

26.

Zhang

Zhu

Wang

, et al. Overexpression of NCAPG inhibits cardia adenocarcinoma apoptosis and promotes epithelial-mesenchymal transition through the Wnt/β-catenin signaling pathway. Gene 2021; 766: 145163.

27.

Wang

Tian

Sui

, et al. Increased expression of NCAPG (Non-SMC condensing I complex subunit G) is associated with progression and poor prognosis of lung adenocarcinoma. Bioengineered 2022; 13: 6113–6125.

28.

Bacalao

Satterthwaite

. Recent advances in lupus B cell biology: PI3K, IFN γ, and chromatin. Front Immunol 2020; 11: 615673.

29.

Xie

Geng

, et al. The multi-targets mechanism of hydroxychloroquine in the treatment of systemic lupus erythematosus based on network pharmacology. Lupus 2020; 29: 1704–1711.

30.

Zhang

Dong

, et al. Network pharmacology modeling identifies synergistic interaction of therapeutic and toxicological mechanisms for Tripterygium hypoglaucum Hutch. BMC Complement Med Ther 2021; 21: 38.

31.

Wang

Xie

Wang

, et al. Azithromycin promotes alternatively activated macrophage phenotype in systematic lupus erythematosus via PI3K/Akt signaling pathway. Cell Death Dis 2018; 9: 1080.

32.

Iacobas

Wen

Iacobas

, et al. TWEAKing the Hippocampus: the effects of TWEAK on the genomic fabric of the Hippocampus in a neuropsychiatric lupus mouse model. Genes 2021; 12: 1172.

33.

Iacobas

Wen

Iacobas

, et al. Remodeling of neurotransmission, chemokine, and PI3K-AKT signaling genomic fabrics in neuropsychiatric systemic lupus erythematosus. Genes 2021; 12: 251.

34.

Liang

Zhang

Qiu

, et al. Dysregulation of tRNA-derived small RNAs and their potential roles in lupus nephritis. Lupus 2021; 30: 2248–2255.