Effect of TUG1 non-coding RNA knockdown on CD20 receptor expression

Rojhannezhad, Mahbubeh; Behmanesh, Mehrdad; Nikravesh, Abas; Naser Moghadasi, Abdolreaza

Effect of TUG1 non-coding RNA knockdown on CD20 receptor expression

Document Type : Original Research

Authors

mahbubeh rojhannezhad ¹

Mehrdad Behmanesh ²

Abas Nikravesh ³

Abdolreaza Naser Moghadasi ⁴

¹ Tarbiat Modares university, Faculty of Biological Sciences, Genetics department, PhD student of Genetics

² Department of Genetics,Faculty of Biological Science, TarbiatModares University, Tehran, Iran

³ Associate Professor of Molecular GeneticsDepartment of Advanced Technologies, School of MedicineNorth Khorasan University of Medical Sciences

⁴ Assistant Professor of NeurologyDepartment of Neurology, School of MedicineMultiple Sclerosis Research CenterSina HospitalTehran University of Medical Sciences

Abstract

Multiple sclerosis (MS) is one of the most common autoimmune diseases in Iran and the world. To date, many drugs have been developed to control the progression of MS as a chronic inflammatory disease of the central nervous system. Rituximab is a chimeric mouse-human monoclonal antibody that binds to the CD20 receptor on the surface of B cells and induces apoptosis. Today, Numerous studies have confirmed the increasing role of non-coding RNAs in regulating the expression of genes and molecular processes, including apoptosis. Furthermore, bioinformatic analysis results indicate that TUG1 LncRNA is differentially expressed in MS patients. Thus, In the present study the possible role of TUG1 in regulating rituximab mechanism of action and apoptosis induction was experimentally investigated. To do this, specific DNAzyme against TUG1 was designed and transfected into Raji cells in the presence or absence of the drug. After transfection, RNA extraction and cDNA synthesis were performed. Then, the expression of target genes was examined by Real-Time PCR technique. The results showed an increase in CD20 expression and a decrease in SMAD2 expression levels. Furthermore, decreased TUG1 gene expression led to an increase in apoptosis and cell accumulation in the G1 phase. It seems that TUG1 expression level can play a significant role in CD20 expression in B cells and therefore on the therapeutic efficacy of rituximab.

Keywords

Rituximab

CD20

long non coding RNA

TUG1

20.1001.1.23222115.1401.13.4.5.4

Subjects

Molecular biotechnology

[1] C. Du and X. Xie, “G protein-coupled receptors as therapeutic targets for multiple sclerosis,” Cell Res., vol. 22, no. 7, pp. 1108–1128, 2012, doi: 10.1038/cr.2012.87.
[2] B. N. Gargari, M. Behmanesh, and M. A. Sahraian, “Effect of vitamin D treatment on interleukin-2 and interleukin-4 genes expression in multiple sclerosis,” Physiol. Pharmacol., vol. 19, no. 1, pp. 14–21, 2015.
[3] K. Cervantes-Gracia and H. Husi, “Integrative analysis of Multiple Sclerosis using a systems biology approach,” Sci. Rep., vol. 8, no. 1, pp. 1–14, 2018, doi: 10.1038/s41598-018-24032-8.
[4] T. Olsson, L. F. Barcellos, and L. Alfredsson, “Interactions between genetic, lifestyle and environmental risk factors for multiple sclerosis,” Nat. Rev. Neurol., vol. 13, no. 1, pp. 26–36, 2016, doi: 10.1038/nrneurol.2016.187.
[5] I. Jelcic et al., “Memory B Cells Activate Brain-Homing, Autoreactive CD4+ T Cells in Multiple Sclerosis,” Cell, vol. 175, no. 1, pp. 85-100.e23, 2018, doi: 10.1016/j.cell.2018.08.011.
[6] A. Abulayha, A. Bredan, H. El Enshasy, and I. Daniels, “Rituximab: Modes of action, remaining dispute and future perspective,” Futur. Oncol., vol. 10, no. 15, pp. 2481–2492, 2014, doi: 10.2217/fon.14.146.
[7] V. F. Cuzzola et al., “Pharmacogenomic update on multiple sclerosis: A focus on actual and new therapeutic strategies,” Pharmacogenomics J., vol. 12, no. 6, pp. 453–461, 2012, doi: 10.1038/tpj.2012.41.
[8] E. Tsareva, O. Kulakova, A. Boyko, and O. Favorova, “Pharmacogenetics of multiple sclerosis: Personalized therapy with immunomodulatory drugs,” Pharmacogenet. Genomics, vol. 26, no. 3, pp. 103–115, 2016, doi: 10.1097/FPC.0000000000000194.
[9] C. Fenoglio et al., “LncRNAs expression profile in peripheral blood mononuclear cells from multiple sclerosis patients,” J. Neuroimmunol., vol. 324, no. August, pp. 129–135, 2018, doi: 10.1016/j.jneuroim.2018.08.008.
[10] Q. M. Liu et al., “Silencing lncRNA TUG1 Alleviates LPS-Induced Mouse Hepatocyte Inflammation by Targeting miR-140/TNF,” Front. Cell Dev. Biol., vol. 8, no. February, pp. 1–11, 2021, doi: 10.3389/fcell.2020.616416.
[11] C. Guo, Y. Qi, J. Qu, L. Gai, Y. Shi, and C. Yuan, “Pathophysiological Functions of the lncRNA TUG1,” Current Pharmaceutical Design, vol. 26, no. 6. pp. 688–700, 2019, doi: 10.2174/1381612826666191227154009.
[12] Y. Li et al., “Effects of complement and serum IgG on rituximab ‑ dependent natural killer cell ‑ mediated cytotoxicity against Raji cells,” pp. 339–347, 2019, doi: 10.3892/ol.2018.9630.
[13] Behmanesh, M ; Moradi, “Evaluation of the role of long noncoding RNAs lnc-DC and TUG1 on the regulation of BDNF expression and cell cycle,” Tarbiat Modares, 1398.
[14] S. T. Livak KJ, “Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method. Methods.,” no. 2001 Dec;25(4):402–8., doi: doi: 10.1006/meth.2001.1262.
[15] N. Nissimov et al., “B cells reappear less mature and more activated after their anti-CD20-mediated depletion in multiple sclerosis,” Proc. Natl. Acad. Sci. U. S. A., vol. 117, no. 41, pp. 25690–25699, 2020, doi: 10.1073/pnas.2012249117.
[16] C. G. Chisari, E. Sgarlata, S. Arena, S. Toscano, M. Luca, and F. Patti, “Rituximab for the treatment of multiple sclerosis: a review,” J. Neurol., vol. 269, no. 1, pp. 159–183, 2022, doi: 10.1007/s00415-020-10362-z.
[17] S. Brancati, L. Gozzo, L. Longo, D. C. Vitale, and F. Drago, “Rituximab in Multiple Sclerosis: Are We Ready for Regulatory Approval?,” Front. Immunol., vol. 12, no. July, 2021, doi: 10.3389/fimmu.2021.661882.
[18] M. S. Czuczman et al., “Acquirement of rituximab resistance in lymphoma cell lines is associated with both global CD20 gene and protein down-regulation regulated at the pretranscriptional and posttranscriptional levels,” Clin. Cancer Res., vol. 14, no. 5, pp. 1561–1570, 2008, doi: 10.1158/1078-0432.CCR-07-1254.
[19] M. R. Smith, “Rituximab ( monoclonal anti-CD20 antibody ): mechanisms of action and resistance,” pp. 7359–7368, 2003, doi: 10.1038/sj.onc.1206939.
[20] J. Edelmann et al., “Rituximab Activates NOTCH1 Signaling in CLL Cells and Induces Changes in the Cytokine Repertoire Favoring a Tumor-Protective Microenvironment,” Blood, vol. 130, no. Supplement 1. pp. 2996–2996, 2017, doi: 10.1182/blood.V130.Suppl_1.2996.2996.
[21] K. Katsushima et al., “Targeting the Notch-regulated non-coding RNA TUG1 for glioma treatment,” Nature Communications, vol. 7. 2016, doi: 10.1038/ncomms13616.
[22] E. b. Zhang et al., “P53-regulated long non-coding RNA TUG1 affects cell proliferation in human non-small cell lung cancer, partly through epigenetically regulating HOXB7 expression.,” Cell Death Dis., vol. 5, pp. 1–12, 2014, doi: 10.1038/cddis.2014.201.
[23] W. Zhong et al., “Increased interleukin-17A levels promote rituximab resistance by suppressing p53 expression and predict an unfavorable prognosis in patients with diffuse large B cell lymphoma,” Int. J. Oncol., vol. 52, no. 5, pp. 1528–1538, 2018, doi: 10.3892/ijo.2018.4299.
[24] D. Wu et al., “Long noncoding RNA TUG1 promotes osteogenic differentiation of human periodontal ligament stem cell through sponging microRNA-222-3p to negatively regulate Smad2/7,” Arch. Oral Biol., vol. 117, 2020, doi: 10.1016/j.archoralbio.2020.104814.
[25] K. C. Kawabata, S. Ehata, A. Komuro, K. Takeuchi, and K. Miyazono, “TGF-β-induced apoptosis of B-cell lymphoma Ramos cells through reduction of MS4A1/CD20,” Oncogene, vol. 32, no. 16, pp. 2096–2106, 2013, doi: 10.1038/onc.2012.219.
[26] H. Zhou, L. Sun, and F. Wan, “Molecular mechanisms of TUG1 in the proliferation, apoptosis, migration and invasion of cancer cells (Review),” Oncol. Lett., vol. 18, no. 5, pp. 4393–4402, 2019, doi: 10.3892/ol.2019.10848.
[27] C. Qin and F. Zhao, “Long non-coding RNA TUG1 can promote proliferation and migration of pancreatic cancer via EMT pathway,” pp. 2377–2384, 2017.
[28] Q. Li et al., “lncRNA TUG1 modulates proliferation, apoptosis, invasion, and angiogenesis via targeting miR-29b in trophoblast cells,” Hum. Genomics, vol. 13, no. 1, p. 50, 2019, doi: 10.1186/s40246-019-0237-z.