OP0008 Deficient autophagy induces chondrocyte dysfunction through lamin a/c accumulation in aging and osteoarthritis

Background Aging-related Osteoarthritis (OA) is characterized by insufficient extracellular matrix synthesis and articular cartilage degradation. Autophagy is essential to maintain chondrocyte homeostasis by regulating the intracellular macromolecule and organelle turnover (1). Previous findings indicated that autophagy is defective in Aging and OA articular cartilage (2,3). However, the specific target/-s that regulates this homeostatic mechanism and affect cartilage integrity are still unknown. Objectives The objective of study is to identify targets regulating autophagy in human chondrocytes. Methods We performed quantitative proteomic analysis of Atg5 knockdown primary uman chondrocytes using iTRAQ (isobaric tags for relative and absolute quantitation) labeling coupled with on-line 2D LC/MS/MS. Protein identification and quantification were performed using Protein Pilot Software v 4.0. Each MS/MS spectrum was searched in the Uniprot/Swissprot database for Homo sapiens . Human chondrocytes and human cartilage from healthy, aged and OA patients were employed to confirm the role of the identified target by Western Blot (WB), Inmunofluorescence (IF) and Inmunohistochemistry (IHC). Importanly, CRISPR/Cas9 genome editing technology was used for mechanism of action studies. Results 24 out of 487 proteins were significantly altered (p Conclusions Lamin A/C, a nuclear protein contributing to structural integrity to the nucleus and matrix was identified as candidate target for regulating cartilage function under defective autophagy, such as aging and OA. These results support the hypothesis that autophagy is decreased with aging. Therefore, targeting Lamin A/C might be a promising strategy to find novel therapeutics for cartilage aging and OA. References Lotz MK and Carames B. Autophagy and cartilage homeostasis mechanisms in joint health, aging and OA. Nat Rev Rheumatol 2011;7:579–587. Carames B, Olmer M, Kiosses WB, Lotz MK. The relationship of autophagy defects to cartilage damage during joint aging in a mouse model. Arthritis Rheumatol 2015;67:1568–76. Acknowledgements This study was supported by Instituto de Salud Carlos III- Ministerio de Economia y Competitividad, Spain-CP11/00095 and Fondo Europeo de Desarrollo Regional (FEDER). Disclosure of Interest None declared