Abstract Rationale Airway basal cells are tissue stem cells that sustain epithelial homeostasis through proliferation and differentiation. Long non-coding RNAs (lncRNAs) regulate key biological processes, including development, differentiation, and cellular senescence, yet the functions of most lncRNAs remain undefined. MIR205HG, an lncRNA that serves as the host transcript for miR-205, is preferentially expressed in airway basal cells. Although recent studies suggest miR-205-independent functions of MIR205HG, its function in airway basal cells remains unresolved. This study aimed to investigate the role of MIR205HG in airway basal cells. Methods MIR205HG was silenced in primary normal human bronchial epithelial (NHBE) cells via siRNA transfection. Gene expression was assessed by qRT-PCR or bulk RNA-seq, and protein levels by immunocytochemistry. Cell proliferation was measured at baseline and 96 h post-transfection. Cellular senescence was evaluated by SA-β-gal staining. For organoid culture, NHBE were transduced with lentiviral shRNA targeting MIR205HG. Results siRNA-mediated MIR205HG silencing reduced its transcript without affecting mature miR-205. Upon MIR205HG knockdown, transcripts of the basal markers KRT5 and KRT14 decreased. Cell proliferation decreased, whereas the proportion of SA-β-gal-positive cells increased. Consistent with cellular senescence, CDKN1A/p21 increased, while LMNB1/Lamin B1 decreased at both the mRNA and protein levels. Moreover, transcripts of IL6 and several MMPs were upregulated, indicating a senescence-associated secretory phenotype (SASP). Bulk RNA-seq revealed downregulation of cell-cycle gene sets and upregulation of inflammation-, collagen-metabolism-, and differentiation-related gene sets. MIR205HG knockdown impaired organoid formation (Figure). Conclusion In vitro, MIR205HG modulates proliferation and differentiation of airway basal cells independently of miR-205; its loss induces cellular senescence and impairs organoid formation, implicating a role in epithelial homeostasis. This abstract is funded by: None
Mori et al. (Fri,) studied this question.