V Deshmukh1, A L O'Green2, C Bossard3, T Seo4, L Lamangan5, M Ibanez6, A Ghias7, C Lai8, L Do9, S Cho10, J Cahiwat11, K Chiu12, M Pedraza13, S Anderson14, R Harris15, L Dellamary16, S Kc17, C Barroga18, B Melchior19, B Tam20, S Kennedy21, J Tambiah22, J Hood23, Y Yazici24. 1. Samumed, LLC, San Diego, CA, USA. Electronic address: vishal@samumed.com. 2. Samumed, LLC, San Diego, CA, USA. Electronic address: alyssao@samumed.com. 3. Samumed, LLC, San Diego, CA, USA. Electronic address: carine@samumed.com. 4. Samumed, LLC, San Diego, CA, USA. Electronic address: tims@samumed.com. 5. Samumed, LLC, San Diego, CA, USA. Electronic address: lisa@samumed.com. 6. Samumed, LLC, San Diego, CA, USA. Electronic address: maureen@samumed.com. 7. Samumed, LLC, San Diego, CA, USA. Electronic address: abdullah@samumed.com. 8. Samumed, LLC, San Diego, CA, USA. Electronic address: carolyn@samumed.com. 9. Samumed, LLC, San Diego, CA, USA. Electronic address: long@samumed.com. 10. Samumed, LLC, San Diego, CA, USA. Electronic address: shawn@samumed.com. 11. Samumed, LLC, San Diego, CA, USA. Electronic address: joec@samumed.com. 12. Samumed, LLC, San Diego, CA, USA. Electronic address: kevin@samumed.com. 13. Samumed, LLC, San Diego, CA, USA. Electronic address: melinda@samumed.com. 14. Samumed, LLC, San Diego, CA, USA. Electronic address: scotta@samumed.com. 15. Samumed, LLC, San Diego, CA, USA. Electronic address: rodney@samumed.com. 16. Samumed, LLC, San Diego, CA, USA. Electronic address: luis@samumed.com. 17. Samumed, LLC, San Diego, CA, USA. Electronic address: sunil@samumed.com. 18. Samumed, LLC, San Diego, CA, USA. Electronic address: charlene@samumed.com. 19. Samumed, LLC, San Diego, CA, USA. Electronic address: benoit@semumed.com. 20. Formerly Samumed, LLC, USA. Electronic address: byytam@sbcglobal.net. 21. Samumed, LLC, San Diego, CA, USA. Electronic address: sarahk@samumed.com. 22. Samumed, LLC, San Diego, CA, USA. Electronic address: jeymi@samumed.com. 23. Formerly Samumed, LLC, USA. Electronic address: hoodpharmaceuticals@gmail.com. 24. Samumed, LLC, San Diego, CA, USA. Electronic address: yusuf@samumed.com.
Abstract
OBJECTIVES: Wnt pathway upregulation contributes to knee osteoarthritis (OA) through osteoblast differentiation, increased catabolic enzymes, and inflammation. The small-molecule Wnt pathway inhibitor, lorecivivint (SM04690), which previously demonstrated chondrogenesis and cartilage protection in an animal OA model, was evaluated to elucidate its mechanism of action. DESIGN: Biochemical assays measured kinase activity. Western blots measured protein phosphorylation in human mesenchymal stem cells (hMSCs), chondrocytes, and synovial fibroblasts. siRNA knockdown effects in hMSCs and BEAS-2B cells on Wnt pathway, chondrogenic genes, and LPS-induced inflammatory cytokines was measured by qPCR. In vivo anti-inflammation, pain, and function were evaluated following single intra-articular (IA) lorecivivint or vehicle injection in the monosodium iodoacetate (MIA)-induced rat OA model. RESULTS: Lorecivivint inhibited intranuclear kinases CDC-like kinase 2 (CLK2) and dual-specificity tyrosine phosphorylation-regulated kinase 1A (DYRK1A). Lorecivivint inhibited CLK2-mediated phosphorylation of serine/arginine-rich (SR) splicing factors and DYRK1A-mediated phosphorylation of SIRT1 and FOXO1. siRNA knockdowns identified a role for CLK2 and DYRK1A in Wnt pathway modulation without affecting β-catenin with CLK2 inhibition inducing early chondrogenesis and DYRK1A inhibition enhancing mature chondrocyte function. NF-κB and STAT3 inhibition by lorecivivint reduced inflammation. DYRK1A knockdown was sufficient for anti-inflammatory effects, while combined DYRK1A/CLK2 knockdown enhanced this effect. In the MIA model, lorecivivint inhibited production of inflammatory cytokines and cartilage degradative enzymes, resulting in increased joint cartilage, decreased pain, and improved weight-bearing function. CONCLUSIONS: Lorecivivint inhibition of CLK2 and DYRK1A suggested a novel mechanism for Wnt pathway inhibition, enhancing chondrogenesis, chondrocyte function, and anti-inflammation. Lorecivivint shows potential to modify structure and improve symptoms of knee OA.
OBJECTIVES: Wnt pathway upregulation contributes to knee osteoarthritis (OA) through osteoblast differentiation, increased catabolic enzymes, and inflammation. The small-molecule Wnt pathway inhibitor, lorecivivint (SM04690), which previously demonstrated chondrogenesis and cartilage protection in an animal OA model, was evaluated to elucidate its mechanism of action. DESIGN: Biochemical assays measured kinase activity. Western blots measured protein phosphorylation in human mesenchymal stem cells (hMSCs), chondrocytes, and synovial fibroblasts. siRNA knockdown effects in hMSCs and BEAS-2B cells on Wnt pathway, chondrogenic genes, and LPS-induced inflammatory cytokines was measured by qPCR. In vivo anti-inflammation, pain, and function were evaluated following single intra-articular (IA) lorecivivint or vehicle injection in the monosodium iodoacetate (MIA)-induced rat OA model. RESULTS:Lorecivivint inhibited intranuclear kinases CDC-like kinase 2 (CLK2) and dual-specificity tyrosine phosphorylation-regulated kinase 1A (DYRK1A). Lorecivivint inhibited CLK2-mediated phosphorylation of serine/arginine-rich (SR) splicing factors and DYRK1A-mediated phosphorylation of SIRT1 and FOXO1. siRNA knockdowns identified a role for CLK2 and DYRK1A in Wnt pathway modulation without affecting β-catenin with CLK2 inhibition inducing early chondrogenesis and DYRK1A inhibition enhancing mature chondrocyte function. NF-κB and STAT3 inhibition by lorecivivint reduced inflammation. DYRK1A knockdown was sufficient for anti-inflammatory effects, while combined DYRK1A/CLK2 knockdown enhanced this effect. In the MIA model, lorecivivint inhibited production of inflammatory cytokines and cartilage degradative enzymes, resulting in increased joint cartilage, decreased pain, and improved weight-bearing function. CONCLUSIONS:Lorecivivint inhibition of CLK2 and DYRK1A suggested a novel mechanism for Wnt pathway inhibition, enhancing chondrogenesis, chondrocyte function, and anti-inflammation. Lorecivivint shows potential to modify structure and improve symptoms of knee OA.
Authors: Tonia L Vincent; Tamara Alliston; Mohit Kapoor; Richard F Loeser; Linda Troeberg; Christopher B Little Journal: Clin Geriatr Med Date: 2022-05 Impact factor: 3.529
Authors: Ning Wang; Yuchen He; Silvia Liu; Meagan J Makarcyzk; Guanghua Lei; Alexander Chang; Peter G Alexander; Tingjun Hao; Anne-Marie Padget; Nuria de Pedro; Tsapekos Menelaos; Hang Lin Journal: Sci China Life Sci Date: 2021-06-04 Impact factor: 6.038