Controlled induction and targeted elimination of p16<sup>INK4a</sup>‐expressing chondrocytes in cartilage explant culture

Wiley - Tập 33 Số 11 - Trang 12364-12373 - 2019
Garrett A. Sessions1, Michaela E. Copp2, Jie‐Yu Liu3, Margaret A. Sinkler4,1, Susan D’Costa1, Brian O. Diekman3,2,1
1Thurston Arthritis Research Center
2Lineberger Comprehensive Cancer Center, the University of North Carolina at Chapel Hill, Chapel Hill, North Carolina, USA
3Joint Department of Biomedical Engineering University of North Carolina at Chapel Hill, Chapel Hill, North Carolina‐North Carolina State University Raleigh North Carolina USA
4Medical College of Georgia, Augusta University, Augusta, Georgia, USA

Tóm tắt

Cellular senescence is a phenotypic state that contributes to age‐related diseases through the secretion of matrix‐degrading and inflammatory molecules. An emerging therapeutic strategy for osteoarthritis (OA) is to selectively eliminate senescent cells by initiating apoptosis. This study establishes a cartilage explant model of senescence induction and senolytic clearance using p16Ink4a expression as a biomarker of senescence. Growth‐factor stimulation of explants increased the expression of p16Ink4a at both the mRNA and protein levels. Applying this culture system to cartilage from p16tdTom reporter mice (a knockin allele with tdTomato fluorescent protein regulated by the endogenous p16Ink4a promoter) demonstrated the emergence of a p16‐high population that was quantified using flow cytometry for tdTomato. Cell sorting was used to separate chondrocytes based on tdTomato fluorescence and p16‐high cells showed higher senescence‐associated β‐galactosidase activity and increased gene expression of the senescence‐associated secretory phenotype as compared with p16‐low cells. The potential for effective senolysis within the cartilage extracellular matrix was assessed using navitoclax (ABT‐263). Navitoclax treatment reduced the percentage of p16‐high cells from 17.9 to 6.1% (mean of 13 matched pairs; P < 0.001) and increased cleaved caspase‐3 confirmed apoptotic activity. Together, these findings establish a physiologically relevant cartilage explant model for testing the induction and elimination of senescent chondrocytes, which will support investigations of senolytic therapy for OA.—Sessions, G. A., Copp, M. E., Liu, J.‐Y., Sinkler, M. A., D'Costa, S., Diekman, B. O. Controlled induction and targeted elimination of p16INK4a‐expressing chondrocytes in cartilage explant culture. FASEB J. 33, 12364–12373 (2019). www.fasebj.org

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Wiley

Tập 33 Số 11

12364-12373

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