Reduced Cathepsin L expression and secretion into the extracellular milieu contribute to lung fibrosis in systemic sclerosis.

Abstract

OBJECTIVES:

Lung fibrosis is the leading cause of death in SSc, with no cure currently available. Antifibrotic Endostatin (ES) production does not reach therapeutic levels in SSc patients, suggesting a deficit in its release from Collagen XVIII by the main cleavage enzyme, Cathepsin L (CTSL). Thus, elucidating a potential deficit in CTSL expression and activity unravels an underlying molecular cause for SSc-driven lung fibrosis.

METHODS:

Fibrosis was induced experimentally using TGF-β in vitro, in primary human lung fibroblasts (pLFs), and ex vivo, in human lung tissues. ES and CTSL expression was quantified using ELISA, RT-qPCR, immunoblotting or immunofluorescence. Recombinant NC1-FLAG peptide was used to assess CTSL cleavage activity. CTSL expression was also compared between SSc vs normal (NL)-derived pLFs and lung tissues.

RESULTS:

ES levels were significantly reduced in media conditioned by TGF-β-induced pLFs. TGF-β-stimulated pLFs significantly reduced expression and secretion of CTSL into the extracellular matrix (ECM). CTSL was also sequestered in its inactive form into extracellular vesicles, further reducing its availability in the ECM. Media conditioned by TGF-β-induced pLFs showed reduced cleavage of NC1-Flag and reduced release of the antifibrotic ES fragment. SSc-derived pLFs and lung tissues expressed significantly lower levels of CTSL compared with NL.

CONCLUSIONS:

Our findings identify CTSL as a protein protective against lung fibrosis via its activation of antifibrotic ES, and whose expression in SSc pLFs and lung tissues is suppressed. Identifying strategies to boost CTSL endogenous levels in SSc patients could serve as a viable therapeutic strategy.

• Authors: Joe E Mouawad, Shailza Sharma, Ludivine Renaud, Joseph M Pilewski, Satish N Nadig, Carol Feghali-Bostwick
• Journal: Rheumatology (Oxford, England) (Rheumatology (Oxford)) Vol. 62 Issue 3 Pg. 1306-1316 (03 01 2023) ISSN: 1462-0332 [Electronic] England
• PMID: 35900152 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
• Copyright: © The Author(s) 2022. Published by Oxford University Press on behalf of the British Society for Rheumatology. All rights reserved. For permissions, please email: [email protected].

Chemical References

• Cathepsin L
• Transforming Growth Factor beta
• CTSL protein, human

Topics

• Humans
• Cathepsin L (metabolism)
• Cells, Cultured
• Fibroblasts (metabolism)
• Fibrosis
• Pulmonary Fibrosis (pathology)
• Scleroderma, Systemic (pathology)
• Skin (pathology)
• Transforming Growth Factor beta (metabolism)