lncRNA NTT/PBOV1 Axis Promotes Monocyte Differentiation and Is Elevated in Rheumatoid Arthritis
Abstract
:1. Introduction
2. Results
2.1. NTT (Noncoding Transcript in T Cells) Is Expressed in Human Monocytic Cells and Is Regulated by C/EBPβ
2.2. NTT Regulates Downstream Gene PBOV-1 via HnRNP-U Binding
2.3. C/EBPβ, NTT, and PBOV1 Expression Levels Were Highly Elevated in Fresh Rheumatoid Arthritis (RA) Patients
2.4. Association of C/EBPβ, NTT, and PBOV1 Expression Levels with RA Disease Severity
2.5. Overexpression of PBOV1 in THP-1 Cells Led to Cell Cycle Arrest and Differentiation to Macrophages
3. Discussion
4. Materials and Methods
4.1. Patient Subjects
4.2. THP-1 Cell Culture, Differentiation, and Polarization Assays
4.3. Isolation of Primary Immune Cells
4.4. Chromatin Immunoprecipitation Assay (ChIP)
4.5. Electroporation for Knockdown and Overexpression Assays
4.6. Reverse-Transcriptase PCR (RT-PCR)
4.7. Cell Cycle Analysis
4.8. Immunofluorescence Staining
4.9. Statistical Analyses
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
Abbreviations
lncRNA | Long noncoding RNA |
PBMC | Peripheral blood mononuclear cells |
RA | Peripheral blood mononuclear cells |
NTT | Noncoding transcript in T cells |
References
- Shi, C.; Pamer, E.G. Monocyte recruitment during infection and inflammation. Nat. Rev. Immunol. 2011, 11, 762–774. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Lawrence, T.; Natoli, G. Transcriptional regulation of macrophage polarization: Enabling diversity with identity. Nat. Rev. Immunol. 2011, 11, 750–761. [Google Scholar] [CrossRef] [PubMed]
- Roberts, C.A.; Dickinson, A.K.; Taams, L.S. The interplay between monocytes/macrophages and CD4(+) T cell subsets in rheumatoid arthritis. Front. Immunol. 2015, 6, 571. [Google Scholar] [CrossRef] [PubMed]
- Roy, S. miRNA in macrophage development and function. Antioxid. Redox Signal. 2016, 25, 795–804. [Google Scholar] [CrossRef] [PubMed]
- Self-Fordham, J.B.; Naqvi, A.R.; Uttamani, J.R.; Kulkarni, V.; Nares, S. MicroRNA: Dynamic regulators of macrophage polarization and plasticity. Front. Immunol. 2017, 8, 1062. [Google Scholar] [CrossRef] [PubMed]
- Zhou, H.; Xiao, J.; Wu, N.; Liu, C.; Xu, J.; Liu, F.; Wu, L. MicroRNA-223 regulates the differentiation and function of intestinal dendritic cells and macrophages by targeting C/EBPβ. Cell Rep. 2015, 13, 1149–1160. [Google Scholar] [CrossRef] [PubMed]
- Simpson, L.J.; Ansel, K.M. MicroRNA regulation of lymphocyte tolerance and autoimmunity. J. Clin. Investig. 2015, 125, 2242–2249. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Chen, Y.G.; Satpathy, A.T.; Chang, H.Y. Gene regulation in the immune system by long noncoding RNAs. Nat. Immunol. 2017, 18, 962–972. [Google Scholar] [CrossRef] [PubMed]
- Heward, J.A.; Lindsay, M.A. Long non-coding RNAs in the regulation of the immune response. Trends Immunol. 2014, 35, 408–419. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Jiang, R.; Tang, J.; Chen, Y.; Deng, L.; Ji, J.; Xie, Y.; Wang, K.; Jia, W.; Chu, W.M.; Sun, B. The long noncoding RNA lnc-EGFR stimulates T-regulatory cells differentiation thus promoting hepatocellular carcinoma immune evasion. Nat. Commun. 2017, 8, 15129. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Mathy, N.W.; Chen, X.M. Long non-coding RNAs (lncRNAs) and their transcriptional control of inflammatory responses. J. Biol. Chem. 2017, 292, 12375–12382. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Murphy, M.B.; Medvedev, A.E. Long noncoding RNAs as regulators of Toll-like receptor signaling and innate immunity. J. Leukoc. Biol. 2016, 99, 839–850. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Zhang, Y.; Cao, X. Long noncoding RNAs in innate immunity. Cell Mol. Immunol. 2016, 13, 138–147. [Google Scholar] [CrossRef] [PubMed]
- Satpathy, A.T.; Chang, H.Y. Long noncoding RNA in hematopoiesis and immunity. Immunity 2015, 42, 792–804. [Google Scholar] [CrossRef] [PubMed]
- Wang, P.; Xue, Y.; Han, Y.; Lin, L.; Wu, C.; Xu, S.; Jiang, Z.; Xu, J.; Liu, Q.; Cao, X. The STAT3-binding long noncoding RNA lnc-DC controls human dendritic cell differentiation. Science 2014, 344, 310–313. [Google Scholar] [CrossRef] [PubMed]
- Wei, S.; Zhao, M.; Wang, X.; Li, Y.; Wang, K. PU.1 controls the expression of long noncoding RNA HOTAIRM1 during granulocytic differentiation. J. Hematol. Oncol. 2016, 9, 44. [Google Scholar] [CrossRef] [PubMed]
- Carpenter, S.; Aiello, D.; Atianand, M.K.; Ricci, E.P.; Gandhi, P.; Hall, L.L.; Byron, M.; Monks, B.; Henry-Bezy, M.; Lawrence, J.B.; et al. A long noncoding RNA mediates both activation and repression of immune response genes. Science 2013, 341, 789–792. [Google Scholar] [CrossRef] [PubMed]
- Li, Z.; Chao, T.C.; Chang, K.Y.; Lin, N.; Patil, V.S.; Shimizu, C.; Head, S.R.; Burns, J.C.; Rana, T.M. The long noncoding RNA THRIL regulates TNFalpha expression through its interaction with hnRNPL. Proc. Natl. Acad. Sci. USA 2014, 111, 1002–1007. [Google Scholar] [CrossRef] [PubMed]
- Amarante, M.K.; de Lucca, F.L.; de Oliveira, C.E.; Pelegrinelli Fungaro, M.H.; Reiche, E.M.; Muxel, S.M.; Ehara Watanabe, M.A. Expression of noncoding mRNA in human blood cells activated with synthetic peptide of HIV. Blood Cells Mol. Dis. 2005, 35, 286–290. [Google Scholar] [CrossRef] [PubMed]
- Liu, A.Y.; Torchia, B.S.; Migeon, B.R.; Siliciano, R.F. The human NTT gene: Identification of a novel 17-kb noncoding nuclear RNA expressed in activated CD4+ T cells. Genomics 1997, 39, 171–184. [Google Scholar] [CrossRef] [PubMed]
- Eshleman, E.M.; Delgado, C.; Kearney, S.J.; Friedman, R.S.; Lenz, L.L. Down regulation of macrophage IFNGR1 exacerbates systemic L. monocytogenes infection. PLoS Pathog. 2017, 13, e1006388. [Google Scholar] [CrossRef] [PubMed]
- Vande Walle, L.; Van Opdenbosch, N.; Jacques, P.; Fossoul, A.; Verheugen, E.; Vogel, P.; Beyaert, R.; Elewaut, D.; Kanneganti, T.D.; Van Loo, G.; et al. Negative regulation of the NLRP3 inflammasome by A20 protects against arthritis. Nature 2014, 512, 69–73. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Pattabiraman, D.R.; Gonda, T.J. Role and potential for therapeutic targeting of MYB in leukemia. Leukemia 2013, 27, 269–277. [Google Scholar] [CrossRef] [PubMed]
- Samusik, N.; Krukovskaya, L.; Meln, I.; Shilov, E.; Kozlov, A.P. PBOV1 is a human de novo gene with tumor-specific expression that is associated with a positive clinical outcome of cancer. PLoS ONE 2013, 8, e56162. [Google Scholar] [CrossRef] [PubMed]
- Pan, T.; Wu, R.; Liu, B.; Wen, H.; Tu, Z.; Guo, J.; Yang, J.; Shen, G. PBOV1 promotes prostate cancer proliferation by promoting G1/S transition. Onco Targets Ther. 2016, 9, 787–795. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- An, G.; Ng, A.Y.; Meka, C.S.; Luo, G.; Bright, S.P.; Cazares, L.; Wright, G.L.J.; Veltri, R.W. Cloning and characterization of UROC28, a novel gene overexpressed in prostate, breast, and bladder cancers. Cancer Res. 2000, 60, 7014–7020. [Google Scholar] [PubMed]
- Wang, L.; Niu, C.H.; Wu, S.; Wu, H.M.; Ouyang, F.; He, M.; He, S.Y. PBOV1 correlates with progression of ovarian cancer and inhibits proliferation of ovarian cancer cells. Oncol. Rep. 2016, 35, 488–496. [Google Scholar] [CrossRef] [PubMed]
- Xin, J.; Li, J.; Feng, Y.; Wang, L.; Zhang, Y.; Yang, R. Downregulation of long noncoding RNA HOTAIRM1 promotes monocyte/dendritic cell differentiation through competitively binding to endogenous miR-3960. Onco Targets Ther. 2017, 10, 1307–1315. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Li, Z.; Li, X.; Jiang, C.; Qian, W.; Tse, G.; Chan, M.T.V.; Wu, W.K.K. Long non-coding RNAs in rheumatoid arthritis. Cell Prolif. 2018, 51, 51. [Google Scholar] [CrossRef] [PubMed]
- Sigdel, K.R.; Cheng, A.; Wang, Y.; Duan, L.; Zhang, Y. The emerging functions of long noncoding rna in immune cells: Autoimmune diseases. J. Immunol. Res. 2015, 2015, 848790. [Google Scholar] [CrossRef] [PubMed]
- Song, J.; Kim, D.; Han, J.; Kim, Y.; Lee, M.; Jin, E.J. PBMC and exosome-derived Hotair is a critical regulator and potent marker for rheumatoid arthritis. Clin. Exp. Med. 2015, 15, 121–126. [Google Scholar] [CrossRef] [PubMed]
- Wu, G.C.; Pan, H.F.; Leng, R.X.; Wang, D.G.; Li, X.P.; Li, X.M.; Ye, D.Q. Emerging role of long noncoding RNAs in autoimmune diseases. Autoimmun. Rev. 2015, 4, 798–805. [Google Scholar] [CrossRef] [PubMed]
- Ruffell, D.; Mourkioti, F.; Gambardella, A.; Kirstetter, P.; Lopez, R.G.; Rosenthal, N.; Nerlov, C. A CREB-C/EBPβ cascade induces M2 macrophage-specific gene expression and promotes muscle injury repair. Proc. Natl. Acad. Sci. USA 2009, 106, 17475–17480. [Google Scholar] [CrossRef] [PubMed]
- Huber, R.; Pietsch, D.; Panterodt, T.; Brand, K. Regulation of C/EBPβ and resulting functions in cells of the monocytic lineage. Cell Signal. 2012, 24, 1287–1296. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Pham, T.H.; Langmann, S.; Schwarzfischer, L.; El Chartouni, C.; Lichtinger, M.; Klug, M.; Krause, S.W.; Rehli, M. CCAAT enhancer-binding protein beta regulates constitutive gene expression during late stages of monocyte to macrophage differentiation. J. Biol. Chem. 2007, 282, 21924–21933. [Google Scholar] [CrossRef] [PubMed]
- Marchese, F.P.; Raimondi, I.; Huarte, M. The multidimensional mechanisms of long noncoding RNA function. Genome Biol. 2017, 18, 206. [Google Scholar] [CrossRef] [PubMed]
- Lu, Y.; Liu, X.; Xie, M.; Liu, M.; Ye, M.; Li, M.; Chen, X.M.; Li, X.; Zhou, R. The NF-κβ-responsive long noncoding RNA FIRRE regulates posttranscriptional regulation of inflammatory gene expression through interacting with hnRNPU. J. Immunol. 2017, 199, 3571–3582. [Google Scholar] [CrossRef] [PubMed]
- Petrovic-Djergovic, D.; Popovic, M.; Chittiprol, S.; Cortado, H.; Ransom, R.F.; Partida-Sanchez, S. CXCL10 induces the recruitment of monocyte-derived macrophages into kidney, which aggravate puromycin aminonucleoside nephrosis. Clin. Exp. Immunol. 2015, 180, 305–315. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Xu, B.; Li, J.; Wu, C.; Liu, C.; Yan, X.; Chang, X. CXCL10 and TRAIL are upregulated by TXNDC5 in rheumatoid arthritis fibroblast-like synoviocytes. J. Rheumatol. 2018, 45, 335–340. [Google Scholar] [CrossRef] [PubMed]
- Zhao, Q.; Kim, T.; Pang, J.; Sun, W.; Yang, X.; Wang, J.; Song, Y.; Zhang, H.; Sun, H.; Rangan, V.; et al. A novel function of CXCL10 in mediating monocyte production of proinflammatory cytokines. J. Leukoc. Biol. 2017, 102, 1271–1280. [Google Scholar] [CrossRef] [PubMed]
- Karin, N.; Razon, H. Chemokines beyond chemo-attraction: CXCL10 and its significant role in cancer and autoimmunity. Cytokine 2018, 109, 24–28. [Google Scholar] [CrossRef] [PubMed]
- Carvalheiro, T.; Horta, S.; Van Roon, J.A.G.; Santiago, M.; Salvador, M.J.; Trindade, H.; Radstake, T.; da Silva, J.A.P.; Paiva, A. Increased frequencies of circulating CXCL10-, CXCL8- and CCL4-producing monocytes and Siglec-3-expressing myeloid dendritic cells in systemic sclerosis patients. Inflamm. Res. 2018, 67, 169–177. [Google Scholar] [CrossRef] [PubMed]
- Lee, J.H.; Kim, B.; Jin, W.J.; Kim, H.H.; Ha, H.; Lee, Z.H. Pathogenic roles of CXCL10 signaling through CXCR3 and TLR4 in macrophages and T cells: Relevance for arthritis. Arthritis Res. Ther. 2017, 19, 63. [Google Scholar] [CrossRef] [PubMed]
Basic Information | Initial Lab Data | Initial Disease Activity | RNA Expression at RA Diagnosis | Clinical Parameters at 2-Year Follow-Up | |||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|---|
Patient No. | Sex | Age | RF (IU/mL) | Anti-CCP (U/mL) | ESR (mm/h) | CRP (mg/dL) | DAS28 | SDAI | C/EBPβ (fold) | NTT (fold) | PBOV1 (fold) | Number of Medications Used in Addition to NSAID | DAS28 |
1 | F | 57 | 28.2 | 29 | 44 | 1.15 | 5.53 | 19.7 | 58.69 | 1379.57 | 46,663.28 | 4 (Hydroxychloroquine, Sulfasalazine, Cyclosporine, Prednisolone) | 3.86 |
2 | F | 56 | 688 | 2.2 | 18 | 0.02 | 4.22 | 15 | 56.69 | 1314.23 | 52,136.28 | NA | NA |
3 | F | 55 | 23.4 | 0.8 | 50 | 4.96 | 5.67 | 12.5 | 25.19 | 1045.52 | 39,786.74 | 3 (Hydroxychloroquine, MTX, Adalimumab) | 1.46 |
4 | F | 25 | 52.3 | 76 | 106 | 2.57 | 4.35 | 7.8 | 14.27 | 1541.37 | 27,554.49 | 3 (Hydroxychloroquine, MTX, Sulfasalazine) | 1.13 |
5 | F | 57 | <20 | 1.4 | 46 | 3.4 | 4.16 | 16 | 12.51 | 522.76 | 8659.09 | 3 (Hydroxychloroquine, Sulfasalazine, Prednisolone) | 2.36 |
6 | F | 66 | 413 | 338 | 53 | 1.81 | 4.92 | 11.2 | 7.75 | 216.77 | 6251.56 | 2 (Hydroxychloroquine, Sulfasalazine) | 2.08 |
7 | F | 34 | 79.8 | 121 | 8 | 0.03 | 3.45 | 8 | 5.68 | 114.56 | 4420.519 | 2 (Hydroxychloroquine, MTX) | 1.74 |
8 | F | 43 | 20 | 3.5 | 28 | 0.68 | 3.46 | 7.1 | 6.52 | 195.36 | 4299.64 | NA | NA |
9 | F | 70 | <20 | 0.7 | 50 | 2.52 | 4.74 | 12.8 | 4.13 | 101.83 | 2033.853 | 0 | 2.67 |
10 | F | 50 | 137 | 10 | 25 | 0.25 | 3.78 | 8 | 4.77 | 163.14 | 6038.51 | 2 (Hydroxychloroquine, MTX) | 2.20 |
© 2018 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
Share and Cite
Yang, C.-A.; Li, J.-P.; Yen, J.-C.; Lai, I.-L.; Ho, Y.-C.; Chen, Y.-C.; Lan, J.-L.; Chang, J.-G. lncRNA NTT/PBOV1 Axis Promotes Monocyte Differentiation and Is Elevated in Rheumatoid Arthritis. Int. J. Mol. Sci. 2018, 19, 2806. https://doi.org/10.3390/ijms19092806
Yang C-A, Li J-P, Yen J-C, Lai I-L, Ho Y-C, Chen Y-C, Lan J-L, Chang J-G. lncRNA NTT/PBOV1 Axis Promotes Monocyte Differentiation and Is Elevated in Rheumatoid Arthritis. International Journal of Molecular Sciences. 2018; 19(9):2806. https://doi.org/10.3390/ijms19092806
Chicago/Turabian StyleYang, Chin-An, Ju-Pi Li, Ju-Chen Yen, I-Lu Lai, Yu-Chen Ho, Yu-Chia Chen, Joung-Liang Lan, and Jan-Gowth Chang. 2018. "lncRNA NTT/PBOV1 Axis Promotes Monocyte Differentiation and Is Elevated in Rheumatoid Arthritis" International Journal of Molecular Sciences 19, no. 9: 2806. https://doi.org/10.3390/ijms19092806