H. Mirazi, S. Wood
University of New England,
United States
Keywords: Microphysiological Systems, Osteoarthritis (OA), Joint on a Chip
Summary:
Tanezumab (RN-624) is a humanized monoclonal antibody that targets nerve growth factor (NGF), a protein known to increase following injury, inflammation, or during chronic pain. After 17 years and $3B USD in development, the drug was rejected by the FDA in the middle of a phase 3 clinical trial due to safety concerns that, despite being investigated as a drug to treat osteoarthritis (OA) pain, it was found to induce rapidly progressing OA in 75% of patients.[1-3] Because it is selective for human NGF, it was not able to be tested preclinically in animals, thus necessitating many early (and expensive) clinical trials for its development. Due to unavailability, preclinical data generated at that time could not include data from in vitro models of multiple joint tissues together that might have predicted the clinical trial outcomes. In this study, the multi-tissue nature of the joint as an organ was recapitulated by co-culturing human osteoblasts, articular chondrocytes, dermal fibroblasts, and THP-1-derived macrophages in Ibidi μ-Slide I Luer chips linked together in series, and the response to tanezumab treatment was assessed to determine if the in vitro co-culture system presented here is capable of detecting cellular responses indicative of OA pathogenesis. To establish a model of healthy joint cell co-culture, cell viability across a 5-day duration using a single shared cell culture media was assessed using multiple assays, including nuclear exclusion-based assessment of viability, PrestoBlue™ assessment of metabolic activity, and lactate dehydrogenase (LDH) cytotoxicity evaluation. To model the OA disease state, macrophages were separately exposed to lipopolysaccharides (LPS) and interferon‐gamma (IFN‐γ) prior to co-culture to induce secretion of inflammatory cytokines relevant to OA pathogenesis. After 24 hrs of co-culture, viability exceeded 83%, metabolic activity increased to >5x monoculture levels, and no measurable increase in LDH release was observed. Secretion of catabolic MMP enzymes and inflammatory cytokines commonly associated with OA were evaluated following up to 5 days of co-culture by utilizing quantitative sandwich ELISA arrays. This work serves as the basis for co-culturing these cells in a biomimetic Microphysiological Articular Joint In a Chip (MAJIC) system being developed the Wood group and commercialized by CellField Technologies, Inc. Dr. Wood is an inventor on a patent related to the technology described in this publication for which he is entitled to receive royalties and/or equity. U.S. Patent No. 12098354B2 was issued to the South Dakota Board of Regents. In addition, Dr. Wood is a partner in a company, CellField Technologies, Inc., that has licensed related technology from the South Dakota Board of Regents. A financial conflict of interest management plan is in place for Dr. Wood through the Office of Sponsored Programs at the University of New England. This material is based upon work supported by National Science Foundation Award 2517512 (2234590) and the Maine Technology Institute.