KRT5⁺/p63⁺ Stem Cells Undergo Senescence in the Human Lung with Pathological Aging

Adult lungs present high cellular plasticity against stress and injury, mobilizing stem/progenitor populations from conducting airways to maintain tissue homeostasis and gas exchange in alveolar spaces. With aging, pulmonary functional and structural deterioration occurs, mainly in pathological conditions, which is associated with impaired stem cell activity and increased senescence in mice. However, the impact of these processes underlying lung physiopathology in relation to aging has not been explored in humans. In this work, we analyzed stem cell (SOX2, p63, KRT5), senescence (p16^INK4A, p21^CIP, Lamin B1) and proliferative (Ki67) markers in lung samples from young and aged individuals, with and without pulmonary pathology. We identified a reduction in SOX2⁺ cells but not p63⁺ and KRT5⁺ basal cells in small airways with aging. In alveoli, we revealed the presence of triple SOX2⁺, p63⁺ and KRT5⁺ cells specifically in aged individuals diagnosed with pulmonary pathologies. Notably, p63⁺ and KRT5⁺ basal stem cells displayed colocalization with p16^INK4A and p21^CIP, as well as with low Lamin B1 staining in alveoli. Further studies revealed that senescence and proliferation markers were mutually exclusive in stem cells with a higher percentage colocalizing with senescence markers. These results provide new evidence of the activity of p63⁺/KRT5⁺ stem cells on human lung regeneration and point out that regeneration machinery in human lung is activated under stress due to aging, but fails to repair in pathological cases, as stem cells would likely become senescent.