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Phenotypic Screening Uncovered Anti-Myocardial Fibrosis Candidates Using a Novel 3D Myocardial Tissue Under Hypoxia

ABSTRACT: Myocardial fibrosis (MF) is a shared hallmark among cardiovascular diseases, reflecting a common thread in their pathogenesis. However, reliable MF models simulating the pathogenesis are limited, which leads to the urgent need for more efficient drugs targeting MF. Herein, a novel contractile three-dimensional (3D) myocardial tissue model integrating cardiomyocytes (CMs), cardiac-fibroblasts (CFs), and bone-marrow-derived macrophages (BMDMs) in collagen hydrogel was developed to simulate the clinical pathological changes of terminal stage of cardiovascular disease, heart failure, and screen compounds with anti-MF activity dependably. The 3D myocardial tissue model demonstrated precise, visualizable, and quantifiable contractile characteristics under hypoxia and drug interventions. 76 compounds extracted from the resins of T.vernicifluum, a traditional Chinese medicine with clear clinical benefits for fibrotic diseases, were screened for anti-fibrotic activity. Following the in vitro assay using 3D oxygen-glucose deprivation (OGD)-treated myocardial tissue model instead of two-dimensional (2D) transforming growth factor-β (TGF-β)-treated CFs model, two candidates including LQ-40 and SQ-3 exerts impressive anti-MF activity, which was further validated in left anterior descending coronary artery (LAD)-MF mouse model. The current results demonstrate the feasibility and advantage of the novel 3D contractile multicellular types model in the discovery of potential drugs for MF, and candidates with promising potential for MF.

ORGANISM(S): Mus musculus Rattus norvegicus

PROVIDER: GSE275028 | GEO | 2025/07/16

REPOSITORIES: GEO

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Project description:MicroRNAs (miRNAs) are a class of highly conserved endogenous noncoding single-stranded small RNAs with a length of 18-22 nucleotides. They are involved in regulation at the posttranscriptional and translational levels through the degradation and translation inhibition of messenger RNA (mRNA). It is estimated that at least 60% of all mammalian genes may be regulated by miRNAs. MiRNAs can help uncover the mechanisms of diseases and provide new entry points into therapy. Accumulated evidence has revealed that miRNAs are involved in the pathological process of cardiovascular disease through specific signaling pathways. To investigate the potential miRNAs for myocardial fibrosis post myocardial infarction, rat models of acute myocardial infarction (AMI) were established by ligating the anterior descending branch of the left coronary artery, while sham-operated rats were only threaded without ligation as a control group. There were three rats in each group. Thirteen differentially expressed miRNAs between the two groups were screened and their expression levels in the model group were all higher than those in the control group. The expression of miR-199a-5p was significantly increased in the model group in qRT-PCR, which was consistent with the results of the gene chip. KEGG enrichment analysis showed that the target genes of miR-199a-5p were enriched in the insulin signaling pathway. Furthermore, dual-luciferase reporter assay indicated that miR-199a-5p could negatively regulate the expression of GSK-3β. After transfection, the expression of miR-199a-5p was increased in the miR-199a-5p mimics group. The protein expression of GSK-3β was decreased in CFs transfected with miR-199a-5p mimics. In summary, our study identified miR-199a-5p could promote the progression of myocardial fibrosis after myocardial infarction by targeting GSK-3β, which provides novel targets for diagnosis and treatment of MF. In summary, our research suggests that miR-199a-5p promotes the progression of myocardial fibrosis after myocardial infarction through the activation of the insulin-PI3K/Akt-GSK-3β signaling pathway.

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Phenotypic Screening Uncovered Anti-Myocardial Fibrosis Candidates Using a Novel 3D Myocardial Tissue Under Hypoxia

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