Keratin 15 protects against cigarette smoke-induced epithelial mesenchymal transformation by MMP-9

Wensi Zhu; Linxiao Han; Yuanyuan Wu; Lin Tong; Ludan He; Qin Wang; Yu Yan; Ting Pan; Jie Shen; Yuanlin Song; Yao Shen; Qiaoliang Zhu; Jian Zhou

doi:10.1186/s12931-023-02598-w

Keratin 15 protects against cigarette smoke-induced epithelial mesenchymal transformation by MMP-9

Respir Res. 2023 Nov 25;24(1):297. doi: 10.1186/s12931-023-02598-w.

Authors

Wensi Zhu^#^{1

2

3}, Linxiao Han^#^{1

2

3}, Yuanyuan Wu^#^{1

2

3}, Lin Tong^#^{1

2

3}, Ludan He^#^{4

2

3}, Qin Wang^{1

2

3}, Yu Yan^{1

2

3}, Ting Pan^{1

2

3}, Jie Shen^{5

6

7}, Yuanlin Song^{4

3}, Yao Shen⁸, Qiaoliang Zhu⁹, Jian Zhou^{10

11

12

13

14

15}

Affiliations

¹ Department of Pulmonary and Critical Care Medicine, Shanghai Respiratory Research Institute, Zhongshan Hospital, Fudan University, 180 Fenglin Rd, Shanghai, 200032, China.
² Shanghai Engineering Research Center of Internet of Things for Respiratory Medicine, 180 Fenglin Road, Shanghai, 200032, China.
³ Shanghai Key Laboratory of Lung Inflammation and Injury, 180 Fenglin Road, Shanghai, 200032, China.
⁴ Shanghai Institute of Infectious Disease and Biosecurity, Fudan University, Shanghai, 200032, China.
⁵ Research Center for Chemical Injury, Emergency and Critical Medicine of Fudan University, Fudan University, Shanghai, 200540, China.
⁶ Center of Emergency and Critical Medicine in Jinshan Hospital of Fudan University, Fudan University, Shanghai, 200540, China.
⁷ Key Laboratory of Chemical Injury, Emergency and Critical Medicine of Shanghai Municipal Health Commission, Shanghai, 200540, China.
⁸ Department of Respiratory and Critical Care Medicine, Shanghai Pudong Hospital, 2800 Gongwei Rd, Shanghai, 201399, China. [email protected].
⁹ Department of Thoracic Surgery, Zhongshan Hospital, Fudan University, 180 Fenglin Road, Shanghai, 200032, China. [email protected].
¹⁰ Department of Pulmonary and Critical Care Medicine, Shanghai Respiratory Research Institute, Zhongshan Hospital, Fudan University, 180 Fenglin Rd, Shanghai, 200032, China. [email protected].
¹¹ Shanghai Engineering Research Center of Internet of Things for Respiratory Medicine, 180 Fenglin Road, Shanghai, 200032, China. [email protected].
¹² Shanghai Key Laboratory of Lung Inflammation and Injury, 180 Fenglin Road, Shanghai, 200032, China. [email protected].
¹³ Research Center for Chemical Injury, Emergency and Critical Medicine of Fudan University, Fudan University, Shanghai, 200540, China. [email protected].
¹⁴ Center of Emergency and Critical Medicine in Jinshan Hospital of Fudan University, Fudan University, Shanghai, 200540, China. [email protected].
¹⁵ Key Laboratory of Chemical Injury, Emergency and Critical Medicine of Shanghai Municipal Health Commission, Shanghai, 200540, China. [email protected].

^# Contributed equally.

Abstract

Background: Chronic obstructive pulmonary disease (COPD), a chronic inflammatory lung disease, is a leading cause of morbidity and mortality worldwide. Prolonged cigarette smoking (CS) that causes irreversible airway remodeling and significantly reduces lung function is a major risk factor for COPD. Keratin15⁺ (Krt15⁺) cells with the potential of self-renewal and differentiation properties have been implicated in the maintenance, proliferation, and differentiation of airway basal cells; however, the role of Krt15 in COPD is not clear.

Methods: Krt15 knockout (Krt15^-/-) and wild-type (WT) mice of C57BL/6 background were exposed to CS for six months to establish COPD models. Krt15-CrePGR;Rosa26-LSL-tdTomato mice were used to trace the fate of the Krt15⁺ cells. Hematoxylin and eosin (H&E) and Masson stainings were performed to assess histopathology and fibrosis, respectively. Furthermore, lentivirus-delivered short hairpin RNA (shRNA) was used to knock down KRT15 in human bronchial epithelial (HBE) cells stimulated with cigarette smoke extract (CSE). The protein expression was assessed using western blot, immunohistochemistry, and enzyme-linked immunosorbent assay.

Results: Krt15^-/- CS mice developed severe inflammatory cell infiltration, airway remodeling, and emphysema. Moreover, Krt15 knockout aggravated CS-induced secretion of matrix metalloproteinase-9 (MMP-9) and epithelial-mesenchymal transformation (EMT), which was reversed by SB-3CT, an MMP-9 inhibitor. Consistent with this finding, KRT15 knockdown promoted MMP-9 expression and EMT progression in vitro. Furthermore, Krt15⁺ cells gradually increased in the bronchial epithelial cells and were transformed into alveolar type II (AT2) cells.

Conclusion: Krt15 regulates the EMT process by promoting MMP-9 expression and protects the lung tissue from CS-induced injury, inflammatory infiltration, and apoptosis. Furthermore, Krt15⁺ cells transformed into AT2 cells to protect alveoli. These results suggest Krt15 as a potential therapeutic target for COPD.

Keywords: Airway remodeling; Chronic obstructive pulmonary disease; Inflammatory infiltration; Matrix metalloproteinase-9.

MeSH terms

Airway Remodeling
Animals
Cigarette Smoking* / adverse effects
Epithelial Cells / metabolism
Epithelial-Mesenchymal Transition / physiology
Humans
Keratin-15 / metabolism
Matrix Metalloproteinase 9 / genetics
Matrix Metalloproteinase 9 / metabolism
Mice
Mice, Inbred C57BL
Nicotiana / toxicity
Pulmonary Disease, Chronic Obstructive* / drug therapy
Pulmonary Disease, Chronic Obstructive* / genetics
Pulmonary Disease, Chronic Obstructive* / prevention & control

Substances

Keratin-15
Matrix Metalloproteinase 9
Mmp9 protein, mouse
Krt15 protein, mouse

Abstract

MeSH terms

Substances

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