Abstract Rationale Injury and repair, a core life sciences topic, underscores high adaptability and self-recovery capacity amid external challenges or internal imbalances. Injurious factors (e.g., tracheobronchial tuberculosis, tracheotomy, endotracheal intubation) induce abnormal airway repair, frequently leading to inflammation and stenosis that compromise patients’ quality of life. Surgical/interventional treatments carry high recurrence, a refractory clinical issue. No data support an airway microbiota-repair association. Through targeted investigation of the airway microbiota, we aim to identify novel therapeutic targets for abnormal post-injury airway repair. Methods Patients were enrolled from the First Affiliated Hospital of Chongqing Medical University and multiple centers, with clinical lavage fluid samples collected meticulously from various airway sites. A total of 150 non-stenotic patients and 150 with benign airway stenosis were included, along with patients with repeated visits at intervals exceeding one month. Samples were transported under strict low-temperature conditions and concentrated via gradient centrifugation. The resuspended bacterial pellets after concentration were preserved for multi-omics sequencing, including 16S rRNA sequencing. Animal experiments such as antibiotic (ABX) depletion and fecal microbiota transplantation (FMT) were conducted for functional validation and mechanistic investigation. Results In patients with airway stenosis, abundances of bacteria Acinetobacter, Sphingomonas, Tardiphaga, Bradyrhizobium,Burkholderia-Caballeronia-Paraburkholderia and Aquabacterium were significantly decreased , while bacteria Heamophilus, Porphyromonas, and Fusobacterium were significantly increased. Microbiota in the stenotic site differed significantly from the stenotic patient’s own non-stenotic site (con1) and normal airways of non-stenotic patients (con2), with distinct shift directions between con1 and con2.Airway microbiota comprises multiple phyla, forming functional modules via dominant positive correlations; Some core genera regulate community stability and interspecific interactions. Microbiota in normal/injured-free airways showed spatial stability. Transient microbes dominated in species count with group-specific abundances; Persistent microbes were more abundant in con1 and con2.Microbiota clustered into 3 clusters: cluster2 prevailed in stenotic patients, while cluster1 and 3 dominated non-stenotic patients. Dominant bacteria and cluster proportions differed between tracheobronchial tuberculosis and other etiologies. Germ-free mice with pseudo-airway injury had more severe damage and slower repair, confirming microbiota’s regulatory role in injury repair. A prediction model based on clusters is planned for clinical use. Conclusion For the first time , this study reveals significant microbiota dysbiosis following airway injury, with the microbial composition exhibiting spatial stability, etiological specificity, and dynamic change characteristics. Core microbial genera and cluster typing serve as potential biomarkers, and prediction models constructed based on clusters are expected to be applied in clinical disease assessment and prognostic prediction of patients with airway stenosis. This abstract is funded by: no
Shi et al. (Fri,) studied this question.