Clinical and Translational Medicine, Год журнала: 2024, Номер 14(3)

Опубликована: Март 1, 2024

Dear Editor, Myocardial fibrosis (MF) is the pathological basis of multiple cardiovascular diseases.1 Heterogeneous nuclear ribonucleoprotein A2/B1 (hnRNPA2B1), a member hnRNP family, participates in transcription and RNA metabolism by RNA-binding. The process includes reading m6A-marked messenger RNAs (mRNAs) mediating microRNA (miRNA) maturation.2 Recently, hnRNPA2B1 was shown to be increased blood acute myocardial infarction patients,3, 4 as well upregulated MF an isoproterenol (ISO)-induced model ISO-treated primary cardiac fibroblasts (Figure S1, S2C–F). In this study, we investigated novel role for MF. knockout (eKO) mice, heart-to-body weight ratio significantly reduced after ISO injection 1B). Echocardiography analysis showed higher LV ejection fraction (%) from 45.93 ± 3.78 61.67 1.51 shortening (FS%, 22.51 2.08 32.03 0.99) eKO mice than wild-type (WT) stimulation 1C,D). HE staining Masson results that deletion ISO-induced inflammatory infiltration collagen deposition 1E,F). Western blotting also confirmed loss expression (CF) biomarkers, including I/III, transforming growth factor beta 1 (TGF-β1) alpha-smooth muscle actin (α-SMA), induced 1G–L). above experimental data suggest improves function suppresses CF mice. Cardiac fibroblasts' proliferation activation are associated with development MF.5 To study whether involved fibroblast activation, knocked down overexpressed pretransfection small interfering (siRNA)-hnRNPA2B1 cDNA plasmids 2A Figure S3A). EdU assay 2B S3B) knockdown decreased EdU+ cells stimulation, while overexpression cells. MTT curve siRNA-hnRNPA2B1+ISO group decreased, however, promoted 2C S3C), which indicate enhances fibroblasts. After extracellular matrix proteins such α-SMA I/III highly expressed, thus, used western detect their or overexpression. HnRNPA2B1 silencing markedly TGF-β1 α-SMA, dramatically these MF-related 2D–H S3D–H). These suggested promotes ISO-stimulated tendency towards fibrosis. Considering can read miRNA mediate maturation, collected on miRNAs regulated hnRNPA2B12 were (Table S1) reported literature. MiR-126, miR-181a, miR-210, miR-99b miR-221 screened out 3A) determined quantitative real-time polymerase chain reaction (qRT-PCR), only three them detected. miR-99b-3p, miR-221-3p miR-210-5p left ventricle more significant 3B). Importantly, elevated miRNA-221-3p blocked compared WT 3C). Moreover, levels 3D). changes at animal cell hnRNPA2B1, selected mechanistic discussion. evaluate CF, transfection mimics inhibitors into performed. Treatment enhanced CF-related genes (Col1a1, Col3a1, Tgfb1 Acta2, 3E–G), retrained those 3H–J). Foxo4, one widely expressed forkhead box (Fox) O family members, regulates oxidative stress/immune response, apoptosis.6 We detected Foxo4 mRNA levels, heart 4A). Subsequently, two binding sites predicted 3′-UTR using StarBase 4B). Thus, speculated could target CF. A luciferase reporter exhibited represses its 3′-UTR. Mutation eliminated repression 4C), confirming direct targeting miR-221-3p. Furthermore, qPCR treatment suppressed 4D), 4E). addition, 4F). Consistent vivo results, 4G). related response.7, 8 evaluated proinflammatory mediators (Il6, Il1b Tnfa). Their ventricles S4A–C). Il6, Tnfa declined under 4H–J). 4K–M). Interestingly, level but did not affect Il6 4N–O S4D,E). confirm hnRNPA2B1-mediated MF, performed rescue assay. hnRNPA2B1-silenced myofibroblasts, numberAs 4P,Q). negative regulation gene Acta2 Tgf1b, 4R–U). indicated key regulator conclusion, our participated mechanism regulating miR-221-3p/Foxo4-mediated response myofibroblast 4V). Xuping Li Shuotao Shi experiments, analyzed wrote manuscript; Zipei Li, Ying Wang Xiaoxiao Qi assisted experiments; Rong Zhang provided technical support review Zhongqiu Liu Yuanyuan Cheng supervised project, edited manuscript. All authors approved final declare no conflict interest. would like thank Guangdong Key Laboratory Translational Cancer Research Chinese Medicine providing platform. This work supported National Natural Science Foundation China (Nos. 82074053, 81930114, 82374070 U22A20368), Province (No. 2019A1515010211), Key-Area Development Program 2020B1111100004) Guangzhou Association Technology Youth Talent Support QT-2023-011). expriments University Animal Care Use Committee number 20221035 IITCM-20190149. Please note: publisher responsible content functionality any supporting information supplied authors. Any queries (other missing content) should directed corresponding author article.

Язык: Английский