Engineered Coiled-Coil Protein for Delivery of Inverse Agonist for Osteoarthritis.
Engineered Coiled-Coil Protein for Delivery of Inverse Agonist for Osteoarthritis.
复制标题
用于传递骨关节炎反向激动剂的工程卷曲螺旋蛋白。
DOI：
10.1021/acs.biomac.8b00158
复制
发表时间：
2018
影响因子：
6.2
通讯作者：
Montclare,JinK
中科院分区：
文献类型：
--
作者：
Yin,Liming;Agustinus,AlbertS;Yuvienco,Carlo;Minashima,Takeshi;Schnabel,NicoleL;Kirsch,Thorsten;Montclare,JinK
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 ofall-transretinoic acid (ATRA) in synovial fluid have been identified as a contributive factor to OA, the synthesis ofde novoantagonists 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 CccSto BMS493 and the stimulator, ATRA, via competitive binding assay, in which ATRA exhibits approximately 5-fold superior association with CccSthan BMS493. Interrogation of the structure of CccSindicates that ATRA causes about 10% loss in helicity, while BMS493 did not impact the structure. Furthermore, CccSself-assembles into nanofibers when bound to BMS493 or ATRA as expected, displaying 11–15 nm in diameter. Treatment of human articular chondrocytesin vitroreveals 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.
