Regulatory role of lncRNA MSTRG.11540.1 on the novel-m1192-3p/CYP2C70 axis in modulating albendazole resistance in Haemonchus contortus

Abstract Background To investigate the role of long non-coding RNAs (lncRNAs) in the regulation of albendazole resistance in Haemonchus contortus (H. contortus) through competing endogenous RNA (ceRNA) mechanisms, a ceRNA network analysis was performed based on RNA-seq data from albendazole-sensitive and albendazole-resistant strains. Starting with significantly upregulated lncRNAs and incorporating functional enrichment analysis of predicted target genes, a potential regulatory axis—MSTRG.11540.1/novel-m1192-3p/HCON_00073890 (CYP2C70)—was identified from the ceRNA network. Transcriptomic analysis revealed that MSTRG.11540.1 and CYP2C70 were significantly upregulated (fold changes of 1.20 and 4.01, respectively; P < 0.01), while novel-m1192-3p was significantly downregulated (fold change: 5.91; P < 0.05) in resistant strains. This study employed dual-luciferase reporter assays and RNA interference (RNAi) to validate the interactions and functional relevance of the candidate RNA molecules. Furthermore, an egg hatch assay (EHA) was conducted to assess the impact of the lncRNA-mediated ceRNA axis on albendazole resistance. The aim was to provide insights into the molecular mechanisms by which lncRNAs may regulate drug resistance in H. contortus. Results A dual-luciferase reporter plasmid containing the predicted binding sites was constructed to investigate the candidate ceRNA axis MSTRG.11540.1/novel-m1192-3p/CYP2C70. The interaction assay revealed that novel-m1192-3p binds to MSTRG.11540.1, but no direct binding was observed between novel-m1192-3p and its predicted target gene CYP2C70. However, overexpression of novel-m1192-3p downregulated the expression of both MSTRG.11540.1 (45.71% reduction compared to NC group, P > 0.05) and CYP2C70 (40.74% reduction versus NC group, P < 0.05). Subsequent RNAi experiments confirmed the existence of the MSTRG.11540.1/novel-m1192-3p/CYP2C70 regulatory axis. Moreover, EHA results showed that knockdown of MSTRG.11540.1 significantly decreased the LD50 value of albendazole (52.91 µg/mL to 32.02 µg/mL). Conclusions This study confirmed that the lncRNA-mediated ceRNA axis MSTRG.11540.1/novel-m1192-3p/CYP2C70 is involved in the regulation of albendazole resistance in H. contortus. These findings provide potential molecular targets for further elucidation of anthelmintic resistance mechanisms and the development of novel therapeutic strategies.