Abstract
BACKGROUND: Tracheal stenosis is a common respiratory disease characterized by abnormal narrowing of the tracheal lumen, which restricts airflow and can lead to symptoms such as dyspnea and even asphyxiation. Although the widespread use of imaging and bronchoscopy has improved diagnostic efficiency, the molecular mechanisms underlying tracheal stenosis remain unclear, particularly regarding the identification of predictive molecular markers. METHODS: This was a retrospective study involving 43 patient samples. A range of statistical methods-including Spearman correlation analysis, multivariable logistic regression, LASSO regression, and ROC curve validation-were employed to systematically screen for molecular targets associated with tracheal stenosis and assess their potential for risk prediction. The study also investigated the roles of ion channels and cell cycle-related molecules through literature analysis. Additionally, a tracheal stenosis animal model was constructed, and RT-qPCR was used to detect differences in mRNA expression levels of candidate molecules (CACNA1D, MKI67, CCNB2, CDKL5, and VAMP2). RESULTS: Respiratory failure and scar diathesis were significantly associated with the occurrence of tracheal stenosis (P < 0.05). Patients with respiratory failure had a notably higher risk of developing tracheal stenosis (P < 0.001), while those with scar diathesis also showed increased susceptibility (P = 0.009). Other clinical characteristics, including age, sex, hypertension, and diabetes, were not significantly associated with stenosis risk (P > 0.05). Regarding molecular markers, positively correlated genes-such as CACNA1D, AMPH, and PTPRD-were expressed at significantly higher levels in patients with tracheal stenosis (P < 0.001), suggesting their role in promoting cell proliferation or ion channel dysfunction. In contrast, negatively correlated markers-such as CALML4, CDKL5, and VAMP2-were expressed at lower levels (P < 0.001), potentially exerting protective effects through inhibition of cell proliferation or maintenance of cellular structure. LASSO regression identified CCNB2, PTPRD, and CACNA1D as positively associated with stenosis risk, whereas VAMP2 and CDKL5 were negatively associated. ROC curve analysis demonstrated excellent model performance (AUC = 0.91), indicating strong predictive capacity of the selected biomarkers. Subgroup analysis showed that elevated MKI67 expression was significantly associated with stenosis risk in patients with respiratory failure, but not in those without. Experimental validation using a tracheal stenosis animal model confirmed the differential expression trends observed in clinical samples: upregulation of CACNA1D, MKI67, and CCNB2, and downregulation of CDKL5 and VAMP2. CONCLUSIONS: This study, for the first time, identified several key molecular targets associated with tracheal stenosis through integrated clinical, computational, and experimental analyses. Dysregulation of ion channel-related genes (e.g., CACNA1D) and cell cycle regulators (e.g., MKI67, CCNB1, CCNB2) may contribute to stenosis by promoting airway smooth muscle proliferation and calcium signaling disturbances. Conversely, protective markers such as CALML4 and CDKL5 may inhibit proliferation and preserve cellular homeostasis. Notably, respiratory failure may amplify the pro-stenotic effects of MKI67 via enhanced inflammation or oxidative stress. The identified markers (e.g., CACNA1D, MKI67) show promise as early predictors for tracheal stenosis, facilitating early screening and risk stratification. Furthermore, targeted therapies-such as calcium channel blockers-against high-risk markers may offer novel treatment options. However, further studies with larger cohorts and mechanistic experiments are warranted to confirm these findings and explore clinical translation.