LncRNA NPTN-IT1-201 Ameliorates Depressive-like Behavior by Targeting miR-142-5p and Regulating Inflammation and Apoptosis via BDNF

Abstract

Objective

Long noncoding RNAs (lncRNAs) and microRNAs (miRNAs) are widely expressed in the brain and are associated with the development of neurological and neurodegenerative diseases. However, their roles and molecular mechanisms in major depressive disorder (MDD) remain largely unknown. This study aimed to identify lncRNAs and miRNAs involved in the development of MDD and elucidate their molecular mechanisms.

Methods

Transcriptome and bioinformatic analyses were performed to identify miRNAs and lncRNAs related to MDD. C57 mice were subjected to chronic unpredictable mild stress (CUMS) to establish a depression model. Lentiviruses containing either lncRNA NPTN-IT1-201 or miR-142-5p were microinjected into the hippocampal region of these mice. Behavioral tests including the sucrose preference test (SPT), tail suspension test (TST), and forced swim test (FST) were conducted to evaluate depressive-like behaviors.

Results

The results revealed that overexpression of lncRNA NPTN-IT1-201 or inhibition of miR-142-5p significantly ameliorated depressive-like behaviors in CUMS-treated mice. Dual-luciferase reporter assays confirmed interactions between miR-142-5p with both brain-derived neurotrophic factor (BDNF) and NPTN-IT1-201. ELISA analysis revealed significant alterations in relevant biomarkers in the blood samples of MDD patients compared to healthy controls. Histological analyses, including HE and Nissl staining, showed marked structural changes in brain tissues following CUMS treatment, which were partially reversed by lncRNA NPTN-IT1-201 overexpression or miR-142-5p inhibition. Immunofluorescence imaging demonstrated significant differences in the levels of BAX, Bcl2, p65, Iba1 among different treatment groups. TUNEL assays confirmed reduced apoptosis in brain tissues following these interventions. Western blotting showed the significant differences in BDNF, BAX, and Bcl2 protein levels among different treatment groups.

Conclusion

NPTN-IT1-201 regulates inflammation and apoptosis in MDD by targeting BDNF via miR-142-5p, making it a potential therapeutic target for MDD.

References

[1] Li S, Li Y, Li X, et al.. Regulatory mechanisms of major depressive disorder risk variants. Mol Psychiatry, 2020, 25(9): 1926-1945

[2] Enoch MA. Genetic and environmental influences on the development of alcoholism: Resilience vs. risk. Ann N Y Acad Sci, 2006, 1094(1): 193-201

[3] Li J, Chen H, Fan F, et al.. White-matter functional topology: A neuromarker for classification and prediction in unmedicated depression. Transl Psychiatry, 2020, 10(1): 365

[4] Zhou H, Zhang Q, Martinez E, et al.. Ketamine reduces aversion in rodent pain models by suppressing hyperactivity of the anterior cingulate cortex. Nat Commun, 2018, 9(1): 3751

[5] Bejarano F, Chang CH, Sun K, et al.. A comprehensive in vivo screen for anti-apoptotic miRNAs indicates broad capacities for oncogenic synergy. Dev Biol, 2021, 475: 10-20

[6] Ghose K, Rai KM. Long noncoding RNAs and their implication in novel trait development in soybean. Long Noncoding RNAs in Plants, 2021 133-150

[7] Li FZ, Zang WQ. Knockdown of lncRNAXLOC_001659 inhibits proliferation and invasion of esophageal squamous cell carcinoma cells. World J Gastroenterol, 2019, 25(42): 6299-6310

[8] Liu J, Yang LZ, Chen LL. Understanding lncRNA–protein assemblies with imaging and single-molecule approaches. Curr Opin Genet Dev, 2022, 72: 128-137

[9] Li M, Xue Y, He R. Isolation of Protein Complexes Associated with Long Noncoding RNAs. Methods Mol Biol, 2021, 2372: 27-33

[10] Xu Y, Cao Z, Ding Y, et al.. Long noncoding RNA NEAT1 alleviates acute-on-chronic liver failure through blocking TRAF6 mediated inflammatory response. Front Physiol, 2019, 10: 1503

[11] Mou SJ, Yang PF, Liu YP, et al.. BCLAF1 promotes cell proliferation, invasion and drug-resistance though targeting lncRNA NEAT1 in hepatocellular carcinoma. Life Sci, 2020, 242: 117177

[12] Pasut A, Matsumoto A, Clohessy JG, et al.. The pleiotropic role of noncoding genes in development and cancer. Curr Opin Cell Biol, 2016, 43: 104-113

[13] Jiang Y, Wang Y, Sun Y, et al.. Long noncoding RNA Peg13 attenuates the sevoflurane toxicity against neural stem cells by sponging microRNA-128-3p to preserve Sox13 expression. PLoS One, 2020, 15(12): e0243644

[14] Keihani S, Kluever V, Fornasiero EF. Brain long noncoding RNAs: Multitask regulators of neuronal differentiation and function. Molecules, 2021, 26(13): 3951

[15] Alural B, Genc S, Haggarty SJ. Diagnostic and therapeutic potential of microRNAs in neuropsychiatric disorders: Past, present, and future. Prog Neuro-psychopharmacol Biol Psychiatry, 2017, 73: 87-103

[16] Liu Z, Li X, Sun N, et al.. Microarray profiling and coexpression network analysis of circulating lncRNAs and mRNAs associated with major depressive disorder. PLoS One, 2014, 9(3): e93388

[17] Cui X, Sun X, Niu W, et al.. Long noncoding RNA: Potential diagnostic and therapeutic biomarker for major depressive disorder. Med Sci Monit, 2016, 22: 5240-5248

[18] Ni X, Liao Y, Li L. Therapeutic role of long noncoding RNA TCONS_00019174 in depressive disorders is dependent on Wnt/β-catenin signaling pathway. J Integr Neurosci, 2018, 17(2): 125-132

[19] Cattaneo A, Bocchio-Chiavetto L, Zanardini R, et al.. BDNF Val66Met polymorphism and protein levels in amniotic fluid. BMC Neurosci, 2010, 11(1): 16

[20] Kerckhove M, Tanaka K, Umehara T, et al.. Targeting miR-223 in neutrophils enhances the clearance of Staphylococcus aureus in infected wounds. EMBO Mol Med, 2018, 10(10): e9024

[21] Frank MG, Baratta MV, Sprunger DB, et al.. Microglia serve as a neuroimmune substrate for stress-induced potentiation of CNS pro-inflammatory cytokine responses. Brain Behav Immun, 2007, 21(1): 47-59

[22] Kreisel T, Frank MG, Licht T, et al.. Dynamic microglial alterations underlie stress-induced depressive-like behavior and suppressed neurogenesis. Mol Psychiatry, 2014, 19(6): 699-709

[23] Willner P. Validity, reliability and utility of the chronic mild stress model of depression: a 10-year review and evaluation. Psychopharmacol (Berl), 1997, 134(4): 319-329

[24] Zhang Y, Du L, Bai Y, et al.. CircDYM ameliorates depressive-like behavior by targeting miR-9 to regulate microglial activation via HSP90 ubiquitination. Mol Psychiatry, 2020, 25(6): 1175-1190

[25] Chen D, Liu G, Xu N, et al.. Knockdown of ARK5 expression suppresses invasion and metastasis of gastric cancer. Cell Physiol Biochem, 2017, 42(3): 1025-1036

[26] Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2–ΔΔCT method. Methods, 2001, 25(4): 402-408

[27] Huang R, Zhang Y, Han B, et al.. Circular RNA HIPK2 regulates astrocyte activation via cooperation of autophagy and ER stress by targeting MIR124-2HG. Autophagy, 2017, 13(10): 1722-1741

[28] Liu L, He L, Yin C, et al.. Effects of palmatine on BDNF/TrkB-mediated trigeminal neuralgia. Sci Rep, 2020, 10(1): 4998

[29] Bose D, Saha P, Mondal A, et al.. Obesity worsens gulf war illness symptom persistence pathology by linking altered gut microbiome species to long-term gastrointestinal, hepatic, and neuronal inflammation in a mouse model. Nutrients, 2020, 12(9): 2764

[30] Xia W, Jie W. ZEB1-AS1/miR-133a-3p/LPAR3/EGFR axis promotes the progression of thyroid cancer by regulating PI3K/AKT/mTOR pathway. Cancer Cell Int, 2020, 20(1): 94

[31] Shih CH, Chuang LL, Tsai MH, et al.. Hypoxia-induced MALAT1 promotes the proliferation and migration of breast cancer cells by sponging MiR-3064-5p. Front Oncol, 2021, 11: 658151

[32] Chen Y, Wei L, Zhang X, et al.. 3 Bromopyruvate sensitizes human breast cancer cells to TRAIL induced apoptosis via the phosphorylated AMPK mediated upregulation of DR5. Oncol Rep, 2018, 40(5): 2435-2444

[33] Wang X, Liu H, Liao X, et al.. Dissecting the roles of LncRNAs in the development of periventricular white matter damage. Front Genet, 2021, 12: 641526

[34] Zhou L, Zhu Y, Chen W, et al.. Emerging role of microRNAs in major depressive disorder and its implication on diagnosis and therapeutic response. J Affect Disord, 2021, 286: 80-86

[35] Gugliandolo A, Silvestro S, Sindona C, et al.. MiRNA: Involvement of the MAPK pathway in ischemic stroke, a promising therapeutic target. Medicina (B Aires), 2021, 57(10): 1053

[36] Cheng D, Logge W, Low JK, et al.. Novel behavioral characteristics of the APP(Swe)/PS1ΔE9 transgenic mouse model of Alzheimer’s disease. Behav Brain Res, 2013, 245: 120-127

[37] Liu S, Liu Q, Ju Y, et al.. Downregulation of miR-383 reduces depression-like behavior through targeting Wnt family member 2 (Wnt2) in rats. Sci Rep, 2021, 11(1): 9223

[38] Liu D, Wang Y, Zhao Y, et al.. LncRNA SNHG5 promotes nasopharyngeal carcinoma progression by regulating miR-1179/HMGB3 axis. BMC Cancer, 2020, 20(1): 178

[39] Yu L, Qu H, Yu Y, et al.. LncRNA-PCAT1 targeting miR-145-5p promotes TLR4-associated osteogenic differentiation of adipose-derived stem cells. J Cell Mol Med, 2018, 22(12): 6134-6147

[40] Wang KC, Chang HY. Molecular mechanisms of long noncoding RNAs. Mol Cell, 2011, 43(6): 904-914

[41] Zhu B, Jin LN, Shen JQ, et al.. Differential expression of serum biomarkers in hemodialysis patients with mild cognitive decline: A prospective single-center cohort study. Sci Rep, 2018, 8(1): 12250

[42] Briken S, Rosenkranz SC, Keminer O, et al.. Effects of exercise on Irisin, BDNF and IL-6 serum levels in patients with progressive multiple sclerosis. J Neuroimmunol, 2016, 299: 53-58

[43] Gezen-Ak D, Dursun E, Hanağası H, et al.. BDNF, TNFα, HSP90, CFH, and IL-10 serum levels in patients with early or late onset alzheimer’s disease or mild cognitive impairment. J Alzheimer’s Dis, 2013, 37(1): 185-195

[44] Alshogran OY, Al-Eitan LN, Altawalbeh SM, et al.. Investigating the contribution of NPSR1, IL-6 and BDNF polymorphisms to depressive and anxiety symptoms in hemodialysis patients. Prog Neuro-psychopharmacol Biol Psychiatry, 2019, 94: 109657

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Huazhong University of Science and Technology