posted on 2018-03-30, 00:00authored byLiming Yin, Albert S. Agustinus, Carlo Yuvienco, Takeshi Minashima, Nicole L. Schnabel, Thorsten Kirsch, Jin K. Montclare
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 (CccS) protein for the encapsulation,
protection, and delivery of BMS493. In this study, we determine the
binding affinity of CccS to BMS493 and the stimulator,
ATRA, via competitive binding assay, in which ATRA exhibits approximately
5-fold superior association with CccS than BMS493.
Interrogation of the structure of CccS indicates
that ATRA causes about 10% loss in helicity, while BMS493 did not
impact the structure. Furthermore, CccS 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 CccS·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.