Hsa_circ_0007460 affects the survival of intracellular Mycobacterium tuberculosis by regulating autophagy and apoptosis of macrophages
Received date: 2023-06-30
Revised date: 2023-08-31
Online published: 2023-09-18
Supported by
National Natural Science Foundation of China(81971900)
Circular RNA (circRNA) is a category of non-coding RNAs characterized by the absence of a 5'-cap and 3′-poly(A) tail, and participates in the physiological processes of various human diseases. Nonetheless, the diagnostic and functional significance of circRNAs in active pulmonary tuberculosis (ATB) remains uncertain. Consequently, the purpose of this study is to investigate whether hsa_circ_0007460 can be employed as a potential diagnostic biomarker in ATB patients and explore its function. The result of real-time quantitative fluorescent PCR (RT-qPCR) validated a notable increase in the expression of hsa_circ_0007460 in the peripheral blood of 32 ATB patients, as well as in THP-1 human macrophages infected with Bacillus Calmette Guerin (BCG) which is an attenuated strain of Mycobacterium bovis. Additionally, the receiver operating curve (ROC) illustrated that the area under the ROC curve (AUC), sensitivity and specificity were 0.7474, 76.67%, and 78.13% respectively. RNase R, Actinomycin D and other experiments confirmed that hsa_circ_0007460 was stabler than its linear mRNA, indicating that hsa_circ_0007460 has potential as a diagnostic biomarker of ATB. Furthermore, Western blot (WB), Cell Counting Kit-8 (CCK-8), plate counting, and immunofluorescence experiments revealed that hsa_circ_0007460 could regulate apoptosis and autophagy of macrophages. The downstream miRNAs and mRNAs were subsequently predicted using bioinformatics, and the hsa circ 0007460/hsa-miR-3127-5p/PATZ1 axis was built. These above results suggest that hsa_circ_0007460 is substantially up-regulated in the peripheral blood of patients with ATB and can be utilized as a potential diagnostic biomarker. In addition, hsa_circ_0007460 can promote apoptosis of macrophages and inhibit autophagy of macrophages, thereby promoting the survival of BCG.
Key words： hsa_circ_0007460; autophagy; apoptosis; TB
Jinyi Zhang, Yumo He, Jingyu Zhou, Shufeng Weng, Huixia Ma, Taiyue Lin, Ying Xu . Hsa_circ_0007460 affects the survival of intracellular Mycobacterium tuberculosis by regulating autophagy and apoptosis of macrophages[J]. Hereditas(Beijing), 2023 , 45(11) : 1039 -1051 . DOI: 10.16288/j.yczz.23-160
| [1] | World Health Organization.Global tuberculosis report 2022. 2022. |
| [2] | Dookie N, Rambaran S, Padayatchi N, Mahomed S, Naidoo K. Evolution of drug resistance in Mycobacterium tuberculosis: a review on the molecular determinants of resistance and implications for personalized care. J Antimicrob Chemother, 2018, 73(5): 1138-1151. |
| [3] | Kaushik AC, Wu QQ, Lin L, Li HB, Zhao LQ, Wen ZL, Song YZ, Wu QH, Wang J, Guo XK, Wang HL, Yu XL, Wei DQ, Zhang SL. Exosomal ncRNAs profiling of mycobacterial infection identified miRNA-185-5p as a novel biomarker for tuberculosis. Brief Bioinform, 2021, 22(6): bbab210. |
| [4] | Memczak S, Jens M, Elefsinioti A, Torti F, Krueger J, Rybak A, Maier L, Mackowiak SD, Gregersen LH, Munschauer M, Loewer A, Ziebold U, Landthaler M, Kocks C, Le Noble F, Rajewsky N. Circular RNAs are a large class of animal RNAs with regulatory potency. Nature, 2013, 495(7441): 333-338. |
| [5] | Liu HJ, Lu G, Wang WX, Jiang XR, Gu SS, Wang J, Yan X, He F, Wang J. A panel of circRNAs in the serum serves as biomarkers for Mycobacterium tuberculosis infection. Front Microbiol, 2020, 11: 1215. |
| [6] | Huang ZK, Su RG, Deng Z, Xu JQ, Peng YP, Luo Q, Li JM. Identification of differentially expressed circular RNAs in human monocyte derived macrophages response to Mycobacterium tuberculosis infection. Sci Rep, 2017, 7(1): 13673. |
| [7] | Zhang XL, Zhang Q, Wu QG, Tang HC, Ye LX, Zhang QL, Hua DM, Zhang YB, Li F. Integrated analyses reveal hsa_circ_0028883 as a diagnostic biomarker in active tuberculosis. Infect Genet Evol, 2020, 83: 104323. |
| [8] | Yuk JM, Yoshimori T, Jo EK. Autophagy and bacterial infectious diseases. Exp Mol Med, 2012, 44(2): 99-108. |
| [9] | Shi HJ, Zhou Y, Jia ET, Liu ZY, Pan M, Bai YF, Zhao XW, Ge QY. Comparative analysis of circular RNA enrichment methods. RNA Biol, 2022, 19(1): 55-67. |
| [10] | Zhai WJ, Wu FJ, Zhang YY, Fu YR, Liu ZJ. The immune escape mechanisms of Mycobacterium tuberculosis. Int J Mol Sci, 2019, 20(2): 340. |
| [11] | Deretic V. Multiple regulatory and effector roles of autophagy in immunity. Curr Opin Immunol, 2009, 21(1): 53-62. |
| [12] | Jayachandran R, Bosedasgupta S, Pieters J. Surviving the Macrophage: Tools and Tricks Employed by Mycobacterium tuberculosis. Curr Top Microbiol Immunol, 2013, 374: 189-209. |
| [13] | Wang Y, Wang SC, Jing HY, Zhang TY, Song N, Xu SW. CircRNA-IGLL1/miR-15a/RNF43 axis mediates ammonia- induced autophagy in broilers jejunum via Wnt/β-catenin pathway. Environ Pollut, 2022, 292(Pt A): 118332. |
| [14] | Wang CS, Chen RS, Zhu XT, Zhang XB.Long non-coding RNAs LINC00689 inhibits the apoptosis of human nucleus pulposus cells via miR-3127-5p/ATG7 axis- mediated autophagy. Open Med (Wars), 2022, 17(1): 1821-1832. |
| [15] | Li J, Han JY, Zhao AM, Zhang GX. CircPAPPA regulates the proliferation, migration, invasion, apoptosis, and cell cycle of trophoblast cells through the miR-3127-5p/ HOXA7 Axis. Reprod Sci, 2022, 29(4): 1215-1225. |
| [16] | Huang YJ, Ren L, Li JJ, Zou HB. Long non-coding RNA PVT1/microRNA miR-3127-5p/NCK-associated protein 1-like axis participates in the pathogenesis of abdominal aortic aneurysm by regulating vascular smooth muscle cells. Bioengineered, 2021, 12(2): 12583-12596. |
| [17] | Yu Z, Wang YZ, Wang B, Zhai JW. Metformin affects paclitaxel sensitivity of ovarian cancer cells through autophagy mediated by long noncoding RNASNHG7/ miR-3127-5p axis. Cancer Biother Radiopharm, 2022, 37(9): 792-801. |
| [18] | Friedman RC, Farh KKH, Burge CB, Bartel DP. Most mammalian mRNAs are conserved targets of microRNAs. Genome Res, 2009, 19(1): 92-105. |
| [19] | Tritz R, Mueller BM, Hickey MJ, Lin AH, Gomez GG, Hadwiger P, Sah DWY, Muldoon L, Neuwelt EA, Kruse CA. siRNA down-regulation of the PATZ1 gene in human glioma cells increases their sensitivity to apoptotic stimuli. Cancer Ther, 2008, 6(B): 865-876. |
| [20] | Fedele M, Crescenzi E, Cerchia L. The POZ/BTB and AT-Hook containing zinc finger 1 (PATZ1) transcription regulator: physiological functions and disease involvement. Int J Mol Sci, 2017, 18(12): 2524. |
| [21] | Ow JR, Ma H, Jean A, Goh Z, Lee YH, Chong YM, Soong R, Fu XY, Yang H, Wu Q. Patz1 regulates embryonic stem cell identity. Stem Cells Dev, 2014, 23(10): 1062-1073. |
| [22] | Fedele M, Franco R, Salvatore G, Paronetto MP, Barbagal F, Pero R, Chiariotti L, Sette C, Tramontano D, Chieffi G, Fusco A, Chieffi P. PATZ1 gene has a critical role in the spermatogenesis and testicular tumours. J Pathol, 2008, 215(1): 39-47. |
| [23] | Valentino T, Palmieri D, Vitiello M, Pierantoni GM, Fusco A, Fedele M. PATZ1 interacts with p53 and regulates expression of p53-target genes enhancing apoptosis or cell survival based on the cellular context. Cell Death Dis, 2013, 4(12): e963. |
| [24] | Guo S, Zhu KX, Yu WH, Wang T, Li S, Wang YX, Zhang CC, Guo JQ. SH3PXD2A-AS1/miR-330-5p/UBA2 ceRNA network mediates the progression of colorectal cancer through regulating the activity of the Wnt/β-catenin signaling pathway. Environ Toxicol, 2021, 36(10): 1969-1980. |
| [25] | Zhuang ZG, Zhang JA, Luo HL, Liu GB, Lu YB, Ge NH, Zheng BY, Li RX, Chen C, Wang X, Liu YQ, Liu FH, Zhou Y, Cai XZ, Chen ZW, Xu JF. The circular RNA of peripheral blood mononuclear cells: hsa_circ_0005836 as a new diagnostic biomarker and therapeutic target of active pulmonary tuberculosis. Mol Immunol, 2017, 90: 264-272. |
| [26] | Huang ZK, Yao FY, Liu J, Xu JQ, Guo Y, Su RG, Luo Q, Li JM.Up-regulation of circRNA-0003528 promotes Mycobacterium tuberculosis associated macrophage polarization via down-regulating miR-224-5p, miR-324-5p and miR-488-5p and up-regulating CTLA4. Aging (Albany NY), 2020, 12(24): 25658-25672. |
| [27] | Luo HL, Pi J, Zhang JA, Yang EZ, Xu H, Luo H, Shen L, Peng Y, Liu GB, Song CM, Li KY, Wu XJ, Zheng BY, Shen HB, Chen ZW, Xu JF.Circular RNA TRAPPC6B inhibits intracellular Mycobacterium tuberculosis growth while inducing autophagy in macrophages by targeting microRNA-874-3p. Clin Transl Immunology, 2021, 10(2): e1254. |
| [28] | Szabo L, Salzman J. Detecting circular RNAs: bioinformatic and experimental challenges. Nat Rev Genet, 2016, 17(11): 679-692. |
| [29] | Deretic V. Autophagy in immunity and cell-autonomous defense against intracellular microbes. Immunol Rev, 2011, 240(1): 92-104. |
| [30] | Gutierrez MG, Master SS, Singh SB, Taylor GA, Colombo MI, Deretic V. Autophagy is a defense mechanism inhibiting BCG and Mycobacterium tuberculosis survival in infected macrophages. Cell, 2004, 119(6): 753-766. |
| [31] | Gajer P, Brotman RM, Bai GY, Sakamoto J, Schütte UME, Zhong X, Koenig SSK, Fu L, Ma ZS, Zhou X, Abdo Z, Forney LJ, Ravel J. Temporal dynamics of the human vaginal microbiota. Sci Transl Med, 2012, 4(132): 132ra52. |
/
| 〈 |
|
〉 |
www.chinagene.cn
备案号：京ICP备09063187号
