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 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 (Ccc S) protein for the encapsulation, protection, and delivery of BMS493. In this study, we determine the binding affinity of Ccc S to BMS493 and the stimulator, ATRA, via competitive binding assay, in which ATRA exhibits approximately 5-fold superior association with Ccc S than BMS493. Interrogation of the structure of Ccc S indicates that ATRA causes about 10% loss in helicity, while BMS493 did not impact the structure. Furthermore, Ccc 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 Ccc 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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Tretinoin
Proteins
Acids
Retinoic Acid Receptors
Interleukin-1beta
Cartilage
Cartilage Oligomeric Matrix Protein
Matrix Metalloproteinase 13
Biochemistry
Protein S
Nanofibers
Drug delivery
Encapsulation
Assays
Peptide Hydrolases
Association reactions
Degradation
Messenger RNA
Fluids

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. (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

Engineered Coiled-Coil Protein for Delivery of Inverse Agonist for Osteoarthritis. / Yin, Liming; Agustinus, Albert S.; Yuvienco, Carlo; Minashima, Takeshi; Schnabel, Nicole L.; Kirsch, Thorsten; Montclare, Jin.

In: Biomacromolecules, Vol. 19, No. 5, 14.05.2018, p. 1614-1624.

Research output: Contribution to journalArticle

Yin, L, Agustinus, AS, Yuvienco, C, Minashima, T, Schnabel, NL, Kirsch, T & Montclare, J 2018, 'Engineered Coiled-Coil Protein for Delivery of Inverse Agonist for Osteoarthritis', Biomacromolecules, vol. 19, no. 5, pp. 1614-1624. https://doi.org/10.1021/acs.biomac.8b00158
Yin L, Agustinus AS, Yuvienco C, Minashima T, Schnabel NL, Kirsch T et al. Engineered Coiled-Coil Protein for Delivery of Inverse Agonist for Osteoarthritis. Biomacromolecules. 2018 May 14;19(5):1614-1624. https://doi.org/10.1021/acs.biomac.8b00158
Yin, Liming ; Agustinus, Albert S. ; Yuvienco, Carlo ; Minashima, Takeshi ; Schnabel, Nicole L. ; Kirsch, Thorsten ; Montclare, Jin. / Engineered Coiled-Coil Protein for Delivery of Inverse Agonist for Osteoarthritis. In: Biomacromolecules. 2018 ; Vol. 19, No. 5. pp. 1614-1624.
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AB - 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 (Ccc S) protein for the encapsulation, protection, and delivery of BMS493. In this study, we determine the binding affinity of Ccc S to BMS493 and the stimulator, ATRA, via competitive binding assay, in which ATRA exhibits approximately 5-fold superior association with Ccc S than BMS493. Interrogation of the structure of Ccc S indicates that ATRA causes about 10% loss in helicity, while BMS493 did not impact the structure. Furthermore, Ccc 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 Ccc 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.

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