Screening and validation of neurogenic bladder miRNA-mRNA regulatory network

doi:10.12307/2023.492

Abstract

Abstract: BACKGROUND: The miRNA-mRNA regulatory network plays an important role in various biological processes, but its specific mechanism in neurogenic bladder after spinal cord injury remains unclear.
OBJECTIVE: To mine the related targets of neurogenic bladder based on bioinformatics and to construct the miRNA-mRNA regulatory network, verified by animal experiments.
METHODS: The differential genes of neurogenic bladder were screened from GEO database to construct the miRNA-mRNA regulatory network. The differential genes were analyzed by gene ontology and Kyoto Encyclopedia of Genes and Genomes analyses using the David database. Protein-protein interaction analysis was performed using the STRING database. Key genes were identified by Cytoscape and ROC curves. Finally the key genes were validated by RT-PCR in a rat model of neurogenic bladder.
RESULTS AND CONCLUSION: A total of 188 differential genes related to neurogenic bladder were identified. Gene ontology and Kyoto Encyclopedia of Genes and Genomes analyses revealed that they were enriched mainly in inflammatory response, cytokine-cytokine receptor interaction, and chemokine signaling pathway. Two key regulatory networks, miR-147-brain-derived neurotrophic factor and miR-362-5p-Scn9a, were identified based on the Cytoscape and ROC curve results. RT-PCR results showed that miR-147, miR-362-5p, and brain-derived neurotrophic factor were highly expressed in the neurogenic bladder rats, while Scn9a was lowly expressed compared with the control rats. Overall, these findings indicate that miR-147-brain-derived neurotrophic factor and miR-362-5p-Scn9a actas important regulators of pathophysiological processes such as inflammation and oxidative stress, which are expected to become new targets for the diagnosis and treatment of neurogenic bladder.

Key words: neurogenic bladder, spinal cord injury, bioinformatics, GEO database, differential genes, regulatory network, miRNA-mRNA, inflammation, oxidative stress

CLC Number:

Guo Shuhui, Yang Ye, Jiang Yangyang, Xu Jianwen. Screening and validation of neurogenic bladder miRNA-mRNA regulatory network[J]. Chinese Journal of Tissue Engineering Research, 2023, 27(20): 3143-3150.

Figures/Tables 14

References

[1] SHANG Z, JIA C, YAN H, et al. Injecting RNA interference lentiviruses targeting the muscarinic 3 receptor gene into the bladder wall inhibits neurogenic detrusor overactivity in rats with spinal cord injury. Neurourol Urodyn. 2019;38(2):615-624.
[2] STOVER SL, DEVIVO MJ, GO BK. History, implementation, and current status of the National Spinal Cord Injury Database. Arch Phys Med Rehabil. 1999;80(11):1365-1371.
[3] 孟祥志,崔慎红,侯晓倩,等.国际国内神经源性膀胱相关研究的可视化分析[J].中国康复理论与实践,2022,28(4):439-446.
[4] LIM V, MAC-THIONG J M, DIONNE A, et al. Clinical Protocol for Identifying and Managing Bladder Dysfunction during Acute Care after Traumatic Spinal Cord Injury. J Neurotrauma. 2021;38(6):718-724.
[5] GUO W, SHAPIRO K, WANG Z, et al. Restoring both continence and micturition after chronic spinal cord injury by pudendal neuromodulation. Exp Neurol. 2021;340:113658.
[6] LIN L, HU K. MiR-147: Functions and Implications in Inflammation and Diseases. Microrna. 2021;10(2):91-96.
[7] SHANG Z, OU T, XU J, et al. MicroRNA expression profile in the spinal cord injured rat neurogenic bladder by next-generation sequencing. Transl Androl Urol. 2020;9(4):1585-1602.
[8] CRUZ C D, COELHO A, ANTUNES-LOPES T, et al. Biomarkers of spinal cord injury and ensuing bladder dysfunction. Adv Drug Deliv Rev. 2015; 82-83:153-159.
[9] KAMBOONLERT K, PANYASRIWANIT S, TANTISIRIWAT N, et al. Effects of Bilateral Transcutaneous Tibial Nerve Stimulation on Neurogenic Detrusor Overactivity in Spinal Cord Injury: A Urodynamic Study. Arch Phys Med Rehabil. 2021;102(6):1165-1169.
[10] GROSS O, LEITNER L, RASENACK M, et al. Detrusor sphincter dyssynergia: can a more specific definition distinguish between patients with and without an underlying neurological disorder? Spinal Cord. 2021;59(9):1026-1033.
[11] BIRKHÄUSER V, LIECHTI MD, ANDERSON CE, et al. TASCI-transcutaneous tibial nerve stimulation in patients with acute spinal cord injury to prevent neurogenic detrusor overactivity: protocol for a nationwide, randomised, sham-controlled, double-blind clinical trial. BMJ Open. 2020;10(8):e039164.
[12] WU SY, JIANG YH, JHANG JF, et al. Inflammation and Barrier Function Deficits in the Bladder Urothelium of Patients with Chronic Spinal Cord Injury and Recurrent Urinary Tract Infections. Biomedicines. 2022;10(2):220.
[13] TOGAN T, AZAP OK, DURUKAN E, et al. The prevalence, etiologic agents and risk factors for urinary tract infection among spinal cord injury patients. Jundishapur J Microbiol. 2014;7(1):e8905.
[14] GONG L, LV Y, LI S, et al. Changes in transcriptome profiling during the acute/subacute phases of contusional spinal cord injury in rats. Ann Transl Med. 2020;8(24):1682.
[15] BAEK A, CHO SR, KIM SH. Elucidation of Gene Expression Patterns in the Brain after Spinal Cord Injury. Cell Transplant. 2017;26(7):1286-1300.
[16] CAO S, YUAN J, ZHANG D, et al. Transcriptome Changes In Dorsal Spinal Cord Of Rats With Neuropathic Pain. J Pain Res. 2019;12:3013-3023.
[17] BELADI RN, VARKOLY KS, SCHUTZ L, et al. Serine Proteases and Chemokines in Neurotrauma: New Targets for Immune Modulating Therapeutics in Spinal Cord Injury. Curr Neuropharmacol. 2021;19(11): 1835-1854.
[18] RUTKOWSKI MD, DELEO JA. The Role of Cytokines in the Initiation and Maintenance of Chronic Pain. Drug News Perspect. 2002;15(10): 626-632.
[19] KIGERL KA, LAI W, WALLACE LM, et al. High mobility group box-1 (HMGB1) is increased in injured mouse spinal cord and can elicit neurotoxic inflammation. Brain Behav Immun. 2018;72:22-33.
[20] NIE H, JIANG Z. Bone mesenchymal stem cell-derived extracellular vesicles deliver microRNA-23b to alleviate spinal cord injury by targeting toll-like receptor TLR4 and inhibiting NF-κB pathway activation. Bioengineered. 2021;12(1):8157-8172.
[21] STIRLING DP, CUMMINS K, MISHRA M, et al. Toll-like receptor 2-mediated alternative activation of microglia is protective after spinal cord injury. Brain. 2014;137(Pt 3):707-723.
[22] STIVERS NS, PELISCH N, OREM BC, et al. The toll-like receptor 2 agonist Pam3CSK4 is neuroprotective after spinal cord injury. Exp Neurol. 2017;294:1-11.
[23] HERMAN P, STEIN A, GIBBS K, et al. Persons with Chronic Spinal Cord Injury Have Decreased Natural Killer Cell and Increased Toll-Like Receptor/Inflammatory Gene Expression. J Neurotrauma. 2018;35(15): 1819-2189.
[24] BUTENSCHöN J, ZIMMERMANN T, SCHMAROWSKI N, et al. PSA-NCAM positive neural progenitors stably expressing BDNF promote functional recovery in a mouse model of spinal cord injury. Stem Cell Res Ther. 2016;7:11.
[25] YOO JM, LEE BD, SOK DE, et al. Neuroprotective action of N-acetyl serotonin in oxidative stress-induced apoptosis through the activation of both TrkB/CREB/BDNF pathway and Akt/Nrf2/Antioxidant enzyme in neuronal cells. Redox Biol. 2017;11:592-529.
[26] CAI H, WANG Y, HE J, et al. Neuroprotective effects of bajijiasu against cognitive impairment induced by amyloid-β in APP/PS1 mice. Oncotarget. 2017;8(54):92621-92634.
[27] TIAN WJ, JEON SH, ZHU GQ, et al. Effect of high-BDNF microenvironment stem cells therapy on neurogenic bladder model in rats. Transl Androl Urol. 2021;10(1):345-355.
[28] VIZZARD MA. Changes in urinary bladder neurotrophic factor mRNA and NGF protein following urinary bladder dysfunction. Exp Neurol. 2000;161(1):273-284.
[29] LI F, WANG X, YANG L. MicroRNA-147 targets BDNF to inhibit cell proliferation, migration and invasion in non-small cell lung cancer. Oncol Lett. 2020;20(2):1931-1937.
[30] LIU G, FRIGGERI A, YANG Y, et al. miR-147, a microRNA that is induced upon Toll-like receptor stimulation, regulates murine macrophage inflammatory responses. Proc Natl Acad Sci U S A. 2009;106(37): 15819-15824.
[31] JIANG C, WU X, LI X, et al. Loss of microRNA-147 function alleviates synovial inflammation through ZNF148 in rheumatoid and experimental arthritis. Eur J Immunol. 2021;51(8):2062-2073.
[32] WU CG, HUANG C. MicroRNA-147 inhibits myocardial inflammation and apoptosis following myocardial infarction via targeting HIPK. Eur Rev Med Pharmacol Sci. 2020; 24(11):6279-6287.
[33] DU Y, YANG F, LV D, et al. MiR-147 inhibits cyclic mechanical stretch-induced apoptosis in L6 myoblasts via ameliorating endoplasmic reticulum stress by targeting BRMS1. Cell Stress Chaperones. 2019; 24(6):1151-1161.
[34] COX JJ, REIMANN F, NICHOLAS AK, et al. An SCN9A channelopathy causes congenital inability to experience pain. Nature. 2006;444(7121): 894-598.
[35] DIB-HAJJ SD, RUSH AM, CUMMINS TR, et al. Gain-of-function mutation in Nav1.7 in familial erythromelalgia induces bursting of sensory neurons. Brain. 2005;128(Pt 8):1847-1854.
[36] NASSAR MA, STIRLING LC, FORLANI G, et al. Nociceptor-specific gene deletion reveals a major role for Nav1.7 (PN1) in acute and inflammatory pain. Proc Natl Acad Sci U S A. 2004;101(34):12706-12711.
[37] WEI X, WANG B, WANG Q, et al. MiR-362-5p, Which Is Regulated by Long Non-Coding RNA MBNL1-AS1, Promotes the Cell Proliferation and Tumor Growth of Bladder Cancer by Targeting QKI. Front Pharmacol. 2020;11: 164.
[38] CHEN Y, LIN L, HU X, et al. Silencing of circular RNA circPDE5A suppresses neuroblastoma progression by targeting the miR-362-5p/NOL4L axis. Int J Neurosci. 2021: 1-11.doi: 10.1080/00207454.2021.1896505.
[39] LIU B, LUO C, LIN H, et al. Long Noncoding RNA XIST Acts as a ceRNA of miR-362-5p to Suppress Breast Cancer Progression . Cancer Biother Radiopharm. 2021;36(6):456-466.