Engineered Coiled-Coil Protein for Delivery of Inverse Agonist for Osteoarthritis

Liming Yin, Albert S. Agustinus, Carlo Yuvienco, Takeshi Minashima, Nicole L. Schnabel, Thorsten Kirsch, Jin K. Montclare

Research output: Contribution to journalArticle

Abstract

Osteoarthritis (OA) results from degenerative and abnormal function of joints, with localized biochemistry playing a critical role in its onset and progression. As high levels of all-trans retinoic acid (ATRA) in synovial fluid have been identified as a contributive factor to OA, the synthesis of de novo antagonists for retinoic acid receptors (RARs) has been exploited to interrupt the mechanism of ATRA action. BMS493, a pan-RAR inverse agonist, has been reported as an effective inhibitor of ATRA signaling pathway; however, it is unstable and rapidly degrades under physiological conditions. We employed an engineered cartilage oligomeric matrix protein coiled-coil (C cc S ) protein for the encapsulation, protection, and delivery of BMS493. In this study, we determine the binding affinity of C cc S to BMS493 and the stimulator, ATRA, via competitive binding assay, in which ATRA exhibits approximately 5-fold superior association with C cc S than BMS493. Interrogation of the structure of C cc S indicates that ATRA causes about 10% loss in helicity, while BMS493 did not impact the structure. Furthermore, C cc S self-assembles into nanofibers when bound to BMS493 or ATRA as expected, displaying 11-15 nm in diameter. Treatment of human articular chondrocytes in vitro reveals that C cc S ·BMS493 demonstrates a marked improvement in efficacy in reducing the mRNA levels of matrix metalloproteinase-13 (MMP-13), one of the main proteases responsible for the degradation of the extracellular cartilage matrix compared to BMS493 alone in the presence of ATRA, interleukin-1 beta (IL-1β), or IL-1 β together with ATRA. These results support the feasibility of utilizing coiled-coil proteins as drug delivery vehicles for compounds of relatively limited bioavailability for the potential treatment of OA.

Original languageEnglish (US)
Pages (from-to)1614-1624
Number of pages11
JournalBiomacromolecules
Volume19
Issue number5
DOIs
StatePublished - May 14 2018

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ASJC Scopus subject areas

  • Bioengineering
  • Biomaterials
  • Polymers and Plastics
  • Materials Chemistry

Cite this

Yin, L., Agustinus, A. S., Yuvienco, C., Minashima, T., Schnabel, N. L., Kirsch, T., & Montclare, J. K. (2018). Engineered Coiled-Coil Protein for Delivery of Inverse Agonist for Osteoarthritis. Biomacromolecules, 19(5), 1614-1624. https://doi.org/10.1021/acs.biomac.8b00158