Regulating effect of miR-132–3p on the changes of MAPK pathway in rat brains and SH-SY5Y cells exposed to excessive fluoride by targeting expression of MAPK1

Article

作者: He, Wen-Wen ; Hong, Wei ; Dong, Yang-Ting ; Xiao, Yan ; Guan, Zhi-Zhong ; Qi, Xiao-Lan ; He, Yan ; Gui, Chuan-Zhi ; Liao, Wei ; Tu, Xi ; Deng, Jie ; Zeng, Xiao-Xiao

BACKGROUNDAlthough the changes of mitogen-activated protein kinase (MAPK) pathway in the central nervous system (CNS) induced by excessive fluoride has been confirmed by our previous findings, the underlying mechanism(s) of the action remains unclear. Here, we investigate the possibility that microRNAs (miRNAs) are involved in the aspect.METHODSAs a model of chronic fluorosis, SD rats received different concentrations of fluoride in their drinking water for 3 or 6 months and SH-SY5Y cells were exposed to fluoride. Literature reviews and bioinformatics analyses were used to predict and real-time PCR to measure the expression of 12 miRNAs; an algorithm-based approach was applied to identify multiply potential target-genes and pathways; the dual-luciferase reporter system to detect the association of miR-132-3p with MAPK1; and fluorescence in situ hybridization to detect miR-132-3p localization. The miR-132-3p inhibitor or mimics or MAPK1 silencing RNA were transfected into cultured cells. Expression of protein components of the MAPK pathway was assessed by immunofluorescence or Western blotting.RESULTSIn the rat hippocampus exposed with high fluoride, ten miRNAs were down-regulated and two up-regulated. Among these, miR-132-3p expression was down-regulated to the greatest extent and MAPK1 level (selected from the 220 genes predicted) was corelated with the alteration of miR-132-3p. Furthermore, miR-132-3p level was declined, whereas the protein levels MAPK pathway components were increased in the rat brains and SH-SY5Y cells exposed to high fluoride. MiR-132-3p up-regulated MAPK1 by binding directly to its 3'-untranslated region. Obviously, miR-132-3p mimics or MAPK1 silencing RNA attenuated the elevated expressions of the proteins components of the MAPK pathway induced by fluorosis in SH-SY5Y cells, whereas an inhibitor of miR-132-3p just played the opposite effect.CONCLUSIONMiR-132-3p appears to modulate the changes of MAPK signaling pathway in the CNS associated with chronic fluorosis.

2024-03-20·Nan fang yi ke da xue xue bao = Journal of Southern Medical University

[MiR-132-3p negatively regulates CAMTA1 to promote Schwann cell proliferation and migration and alleviates I-125 seeds-induced exacerbation of facial nerve injury in rats].

Article

作者: Ouyang, X ; Shi, Y ; Liu, Y ; Xia, B ; Qian, Y ; Zhu, J ; Yu, L

OBJECTIVETo investigate the regulatory effect of miR-132-3p on calmodulin-binding transcription activator 1 (CAMTA1) and Schwann cell activity in rats with facial nerve injury (FNI) treated with I-125 seeds.METHODSRat Schwann cells were irradiated with I-125 seeds and transfected with miR-132-3p mimic, miR-132-3p inhibitor or sh-CAMTA1. The expressions of S100B and β-tubulin Ⅲ in the cells were detected with immunofluorescence assay, and the expressions of miR-132-3p and CAMTA1 protein were determined using RT-qPCR and Western blotting, respectively. EdU staining and Transwell assay were used to evaluate the changes in cell proliferation and migration ability. In a rat model of FNI, I-125 seeds were implanted into the facial tissues near the facial nerve 2 weeks before modeling, and miR-132-3p mimic was injected subcutaneously in the face after modeling. The pathologies of the facial nerve was assessed by HE, LFB and immunofluorescence staining. The targeting relationship between miR-132-3p and CAMTA1 was verified using StarBase v2.0 database and dual-luciferase reporter assay.RESULTSRat Schwann cells showed high expressions of S100B and β-tubulin Ⅲ. I-125 seeds radiation significantly decreased miR-132-3p expression and repressed proliferation and migration of the cells (P < 0.001). Overexpression of miR-132-3p or CAMTA1 knockdown obviously enhanced proliferation and migration of the Schwann cells, while miR-132-3p knockdown produced the opposite effect. MiR-132-3p negatively regulated CAMTA1 expression. In the rat models of FNI, miR-132-3p injection significantly inhibited CAMTA1 expression and attenuated I-125 seeds-induced exacerbation of FNI.CONCLUSIONOverexpression of miR-132-3p suppresses CAMTA1 expression and promotes Schwann cell proliferation and migration to alleviate I-125 seeds-induced exacerbation of FNI in rats.

2024-01-01·Molecular Metabolism

Lysine tRNA fragments and miR-194-5p co-regulate hepatic steatosis via β-Klotho and perilipin 2

Article

作者: Tzur, Yonat ; Dubnov, Serafima ; Hanin, Geula ; Gammal, Asaad ; Paldor, Iddo ; Arkin, Isaiah T ; Tam, Joseph ; Greenberg, David S ; Soreq, Hermona ; Winek, Katarzyna ; Madrer, Nimrod ; Bennett, Estelle R

OBJECTIVENon-alcoholic fatty liver disease (NAFLD) involves hepatic accumulation of intracellular lipid droplets via incompletely understood processes. Here, we report distinct and cooperative NAFLD roles of LysTTT-5'tRF transfer RNA fragments and microRNA miR-194-5p.METHODSCombined use of diet induced obese mice with human-derived oleic acid-exposed Hep G2 cells revealed new NAFLD roles of LysTTT-5'tRF and miR-194-5p.RESULTSUnlike lean animals, dietary-induced NAFLD mice showed concurrent hepatic decrease of both LysTTT-5'tRF and miR-194-5p levels, which were restored following miR-132 antisense oligonucleotide treatment which suppresses hepatic steatosis. Moreover, exposing human-derived Hep G2 cells to oleic acid for 7 days co-suppressed miR-194-5p and LysTTT-5'tRF levels while increasing lipid accumulation. Inversely, transfecting fattened cells with a synthetic LysTTT-5'tRF mimic elevated mRNA levels of the metabolic regulator β-Klotho while decreasing triglyceride amounts by 30% within 24 h. In contradistinction, antisense suppression of miR-194-5p induced accumulation of its novel target, the NAFLD-implicated lipid droplet-coating PLIN2 protein. Further, two out of 15 steatosis-alleviating screened drug-repurposing compounds, Danazol and Latanoprost, elevated miR-194-5p or LysTTT-5'tRF levels.CONCLUSIONOur findings highlight the different yet complementary roles of miR-194-5p and LysTTT-5'tRF and offer new insights into the complex roles of small non-coding RNAs and the multiple pathways involved in NAFLD pathogenesis.

项与 miR-132 antisense oligonucleotide inhibitor (Regulus Therapeutics) 相关的新闻（医药）

2022-09-08

·BioSpace

Cardior Announces CE Marking of CardiorHealth miR-132 PCR Kit Used in Ongoing HF-REVERT Phase 2 Clinical Study

HANOVER, Germany--(BUSINESS WIRE)-- Cardior Pharmaceuticals, a clinical-stage biotech company developing non-coding RNA (ncRNA)-based therapeutics for patients with cardiac diseases, today announced the CE marking of its PCR kit which measures circulating microRNA-132 (miR-132) levels in the blood of patients receiving the company’s lead candidate CDR132L, an oligonucleotide-based miR-132 inhibitor. The company is currently conducting its Phase 2 clinical study, HF-REVERT (NCT05350969), to evaluate the safety and efficacy of CDR132L in patients with heart failure and reduced left ventricular ejection fraction (LVEF) after a myocardial infarction. By using the CardiorHealth miR-132 PCR kit, Cardior ensures consistent measurement of miR-132 levels following CDR132L treatment across all clinical trial centers as an indicator of the target engagement and the compound’s mechanism of action. Cardior is developing CDR132L as a therapeutic candidate to improve cardiac systolic and diastolic function in patients with heart failure. By selectively blocking aberrant miR-132 levels, CDR132L has the potential to reverse cellular pathology and restore normal cardiac function. The CardiorHealth miR-132 PCR kit is the first PCR kit designed to monitor the treatment with a non-coding RNA-based therapeutic. “Receiving the CE mark for our CardiorHealth miR-132 Plasma PCR kit is another demonstration of our ongoing commitment to pioneering a paradigm shift in the treatment of cardiac disease,” said Prof. Dr. Dr. Thomas Thum, CSO and Founder of Cardior. “The kit allows us to monitor the effect of our lead candidate in patients and provides us and participating clinicians with valuable information at a high level of certainty.” The blocking of abnormal miR-132 levels by CDR132L as measured with the PCR kit will be an exploratory parameter in the Phase 2 HF-REVERT trial. CE mark approval of the kit was based on the clinical evaluation of the test in patients receiving various doses of CDR132L or placebo, where the kit demonstrated comparable results to a similar testing protocol used in Cardior’s Phase 1b clinical study. The CE mark confirms that Cardior’s proprietary kit meets the essential requirements of the European in vitro diagnostic regulation (EU) 2017/746 for its intended use. *** About HF-REVERT HF-REVERT (NCT05350969) is a multicenter, randomized, parallel, three-arm, placebo-controlled clinical proof-of-concept study designed to evaluate the safety and efficacy of CDR132L in 280 patients with heart failure and reduced left ventricular ejection fraction (LVEF) after a myocardial infarction. The primary endpoint of the study is defined as percentage change from baseline in the left ventricular end-systolic volume assessed by echocardiography (ECHO). Patients enrolled in the Phase 2 study will be randomized to receive intravenous CDR132L infusions with either 5 mg/kg or 10 mg/kg or placebo, administered 28 days apart as an add-on to Standard of Care (SoC) treatment. The study will be conducted at approximately 60 clinical study centers across Europe. About CDR132L CDR132L is a synthetic antisense oligonucleotide-based inhibitor directed against the microRNA-132 (miR-132). miR-132 is up-regulated in cardiac tissue of heart failure patients causing an aberrant expression of genes that are crucially involved in cardiac function and lead to pathological cardiac remodeling. By blocking miR-132 with its selective inhibitor CDR132L, Cardior is able to restore healthy levels of miR-132 and trigger a concerted therapeutic effect against key hallmarks of heart disease including pathological remodeling processes, impaired contractility and fibrosis. CDR132L has the potential to restore heart function thereby prolonging the patient’s life span as well as improving quality of life. The candidate is currently under evaluation in the company’s Phase 2 HF-REVERT trial and has successfully completed Phase 1b clinical studies demonstrating a favorable safety pro beneficial cardiac effects in heart failure patients. Results were published in the European Heart Journal in 2021. About Cardior Cardior Pharmaceuticals is a leading clinical-stage biopharmaceutical company pioneering the discovery and development of RNA-based therapeutics designed to prevent, repair and reverse diseases of the heart. Cardior’s therapeutic approach uses distinctive non-coding RNAs as an innovative platform for addressing the root causes of cardiac dysfunctions. The company aspires to bring transformative therapeutics and diagnostics to patients and thereby make a lasting impact on the treatment of cardiac diseases worldwide. View source version on businesswire.com: Contacts Contact for Cardior Dr. Claudia Ulbrich / Eva Straßburg Cardior Pharmaceuticals GmbH Phone: +49 (0) 511 33 85 99 30 info@cardior.de Media Inquiries Trophic Communications Eva Mulder Phone: +31 6 52 33 15 79 cardior@trophic.eu Source: Cardior Pharmaceuticals View this news release online at:

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100 项与 miR-132 antisense oligonucleotide inhibitor (Regulus Therapeutics) 相关的药物交易

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