miR-146a-3p as a potential novel therapeutic by targeting MBD2 to mediate Th17 differentiation in Th17 predominant neutrophilic severe asthma

Abstract

Th17 (T-helper 17) cells subtype of non-T2 (non-type 2) asthma is related to neutrophilic infiltration and resistance to inhaled corticosteroids (ICS), so is also known as severe asthma. Methyl-CpG binding domain protein 2 (MBD2) regulates the differentiation of the Th17 cells, tending to show a therapeutic target in severe asthma. miR-146a-3p is associated with anti-inflammatory characteristics and immunity. Moreover, bioinformatic analysis showed that MBD2 may be a target gene of miR-146a-3p. However, the role of miR-146a-3p in the differentiation of Th17 cells via MBD2 in severe asthma remains unknown. Here, we aimed to explore how miR-146a-3p interacts with MBD2 and affects the differentiation of Th17 cells in severe asthma. First, we recruited 30 eligible healthy people and 30 patients with severe asthma to detect the expression of miR-146a-3p in peripheral blood mononuclear cells (PBMCs) by qRT-PCR. Then, we established a HDM/LPS (house dust mite/lipopolysaccharide) exposure model of bronchial epithelial cells (BECs) to evaluate the expression of miR-146a-3p, the interaction between miR-146a-3p and MBD2 using western blot and luciferase reporter analysis and the effect of miR-146a-3p regulated Th17 cells differentiation by flow cytometry in BECs in vitro. Finally, we constructed a mouse model of Th17 predominant neutrophilic severe asthma to assess the therapeutic potential of miR-146a-3p in severe asthma and the effect of miR-146a-3p regulated Th17 cells differentiation via MBD2 in vivo. Decreased miR-146a-3p expression was noted in severe asthma patients, in the BECs and in the animal severe asthma models. Moreover, we demonstrated that miR-146a-3p suppressed Th17 cells differentiation by targeting the MBD2. miR-146a-3p overexpression significantly reduced airway hyperresponsiveness, airway inflammation and airway mucus secretion, while also inhibiting Th17 cells response in vivo, which relieved severe asthma. By targeting MBD2 to suppress Th17 cells differentiation, miR-146a-3p provides a potential novel therapeutic for Th17 predominant neutrophilic severe asthma.

miRNAs as Modern Biomarkers in Asthma Therapy

“…The above relationships were simultaneously confirmed in mouse lung tissue. Inhalation of the mimic equivalent of miRNA-146a-3p results in a decrease in MBD2 and the number of Th17 lymphocytes involved in the inflammatory process, inhibition of mucus production, and airway hyperreactivity [67]. In the group of 68 pediatric patients with asthma, the reduction of miRNA-135b and the relationship of this molecule with the increase in CXCL12 (C-X-C motif chemokine ligand 12) were confirmed.…”

Identification of miRNAs biomarkers for prostate cancer in serum and tissue based on bioinformatic analysis and experimental validation

“…Two miRNAs, miR-146a-3p and miR-136-3p, both downregulated in serum and tissue were identi ed. Recent research suggested that miR-146a-3p is related to the occurrence and progression of disease including asthma, allergic rhinitis and Paget's disease (Duan et al 2023, Xia et al 2023, Stephens et al 2020). Similarly, miR-136-3p is also reported to inhibit tumorigenesis (Xu 2020).…”

Combination of androgen and estrogen improves asthma by mediating Runx3 expression

“…DHT and E2 (Apex BIO) were first dissolved with 2mL of DMSO (Sigma Aldrich) first and then diluted with corn oil to a final concentration of 2mg/mL in accordance with the manufacturer's protocol. The dosage of DHT and/or E2 (30mg/kg) was adopted in accordance to previous reports with minor modification 14 , 21 . On day 21, mice were anesthetized with intraperitoneal injection of 10% chloral hydrate and northen euthanized with cervical dislocation maneuver.…”