An in vivo screening platform identifies senolytic compounds that target p16INK4a+ fibroblasts in lung fibrosis

• Published in: The Journal of clinical investigation Vol. 134; no. 9; pp. 1 - 16
• Main Authors: Lee, Jin Young, Reyes, Nabora S, Ravishankar, Supriya, Zhou, Minqi, Krasilnikov, Maria, Ringler, Christian, Pohan, Grace, Wilson, Chris, Ang, Kenny Kean-Hooi, Wolters, Paul J, Tsukui, Tatsuya, Sheppard, Dean, Arkin, Michelle R, Peng, Tien
• Format: Journal Article
• Language: English
• Published: United States American Society for Clinical Investigation 07.03.2024
• Subjects: Age; Animal models; Biomarkers; Cell lines; Cells; Fibroblasts; Fibrosis; Gene expression; High-throughput screening; Hsp90 protein; INK4a protein; Lung diseases; Lungs; p16 Protein; Pulmonary fibrosis; Senescence; Pulmonology; Aging; Cellular senescence; Fibrosis; Drug screens
• ISSN: 1558-8238; 0021-9738; 1558-8238
• DOI: 10.1172/JCI173371

The appearance of senescent cells in age-related diseases has spurred the search for compounds that can target senescent cells in tissues ("senolytics"). However, a major caveat with current senolytic screens is the use of cell lines as targets where senescence is induced in vitro, which does not necessarily reflect the identity and function of pathogenic senescent cells in vivo. Here, we developed a new pipeline leveraging a fluorescent murine reporter that allows for isolation and quantification of p16Ink4a+ cells in diseased tissues. By high-throughput screening in vitro, precision cut lung slice (PCLS) screening ex vivo, and phenotypic screening in vivo, we identified a HSP90 inhibitor (XL888) as a potent senolytic in tissue fibrosis. XL888 treatment eliminated pathogenic p16Ink4a+ fibroblasts in a murine model of lung fibrosis and reduced fibrotic burden. Finally, XL888 preferentially targeted p16INK4a-high human lung fibroblasts isolated from patients with idiopathic pulmonary fibrosis (IPF), and reduced p16INK4a+ fibroblasts from IPF PCLS ex vivo. This study provides proof of concept for a platform where p16INK4a+ cells are directly isolated from diseased tissues to identify compounds with in vivo and ex vivo efficacy in mouse and human respectively and provides a senolytic screening platform for other age-related diseases.