Fu Shu,1,2 Yaping Wang,3 Yiheng Jiang,4 Linglong Li,3 Zengyi Mu,5 Lei Shi,5 Xiaobao Gong,2 Baoshun Zhang,2 Feng Zhang,3 Dehong Mao3 

1College of Traditional Chinese Medicine, Chongqing Medical University, Chongqing, 400000, People’s Republic of China; 2College of Pharmaceutical Sciences, Southwest University, Chongqing, 400000, People’s Republic of China; 3Department of Otorhinolaryngology, Yongchuan Chinese Medicine Hospital Affiliated to Chongqing Medical University, Chongqing, 400000, People’s Republic of China; 4Department of Laboratory Medicine, West China Hospital, Sichuan University, Chengdu, 610000, People’s Republic of China; 5Department of Otorhinolaryngology, The Affiliated Hospital of Liaoning University of Traditional Chinese Medicine, Shenyang, 110847, People’s Republic of China

Correspondence: Dehong Mao, Yongchuan Chinese Medicine Hospital Affiliated to Chongqing Medical University, No. 2 Yingbin Avenue, Yongchuan District, Chongqing, 400000, People’s Republic of China, Tel +86-023-49822235, Email dehong1963@163.com Feng Zhang, Yongchuan Chinese Medicine Hospital Affiliated to Chongqing Medical University, No. 2 Yingbin Avenue, Yongchuan District, Chongqing, 400000, People’s Republic of China, Tel +86-023-49815555, Email Zhangfeng164382044@163.com

Purpose: Type 2 inflammation and epithelial-mesenchymal transition (EMT) are critical components in the pathogenesis of eosinophilic chronic rhinosinusitis (ECRS), yet their upstream regulatory mechanisms remain poorly understood. This study aimed to explore the regulatory role of miR-301b-3p in these mechanisms and evaluate its therapeutic potential.
Methods: High-throughput miRNA sequencing of human and mouse nasal mucosa tissues identified miR-301b-3p as a key candidate molecule. Mendelian randomization (MR) analysis confirmed its causal relationship with chronic rhinosinusitis (CRS). The role of miR-301b-3p and its target gene PIK3CB in ECRS was further investigated using bioinformatics analysis, dual-luciferase reporter assays, adeno-associated virus (AAV)-mediated modulation of miR-301b-3p expression, histological staining, and a range of molecular biology techniques to elucidate the underlying mechanisms. The role of PIK3CB was assessed using the PIK3CB inhibitor TGX-221.
Results: hsa-miR-301b-3p was significantly downregulated in ECRS patient nasal mucosa (log2FC = – 1.636, P = 0.010), and its expression level negatively correlated with disease severity; simultaneously, mmu-miR-301b-3p was significantly downregulated in the ECRS mouse model (log2FC = – 2.256, P = 0.041). MR analysis demonstrated a causal relationship between reduced miR-301b levels and increased CRS risk (OR = 0.956; 95% CI, 0.918– 0.996; P = 0.033). In vivo experiments revealed that miR-301b-3p knockdown led to PIK3CB upregulation and activation of the PI3K-AKT pathway, triggering type 2 inflammation and EMT. Conversely, overexpression of miR-301b-3p suppressed PIK3CB expression and mitigated these pathological changes. Notably, the PIK3CB inhibitor TGX-221 reversed PI3K-AKT hyperactivation induced by miR-301b-3p knockdown, alleviating type 2 inflammation and EMT.
Conclusion: miR-301b-3p regulates type 2 inflammation and EMT in ECRS by targeting PIK3CB and modulating the PI3K-AKT pathway, suggesting both miR-301b-3p and PIK3CB as promising therapeutic targets. Given the limited sample size of this study, we plan to expand our cohort and implement a longitudinal follow-up design to monitor dynamic changes in miR-301b-3p expression and their relationship to disease progression.

Keywords: miR-301b-3p, PIK3CB, eosinophilic chronic rhinosinusitis, PI3K-AKT pathway, epithelial-mesenchymal transition