Session Type: Abstract Submissions (ACR)
Background/Purpose 14-3-3 proteins represent a highly conserved seven-member family of ubiquitously expressed intracellular chaperonins that perform a broad range of signaling functions. The 14-3-3 eta (η) protein is an emerging diagnostic and prognostic biomarker for RA driving inflammation and joint erosion.1 Elevated levels of the η isoform have been reported in synovial fluid and serum from patients with joint inflammation2, but not with other diseases including psoriasis, osteoporosis, SLE, Crohn’s and MS. Here we evaluate the impact of this RA-associated biomarker on the levels of clinically relevant biomarkers across a panel of human primary cell-based disease models in the BioMAP®platform.
Methods BioMAP® systems model disease biology in primary human cells cultured alone or with different stimulus combinations and have been used extensively to characterize compounds based on phenotypic signatures.2,3Phenotypic activity profiles were generated for eight concentrations of 14-3-3η observed in RA patients (0.25-50 ng/ml) across a panel of 13 BioMAP systems modeling vascular, immune, inflammation and tissue remodeling biology relevant for various human diseases. Changes in protein-based, clinically relevant endpoints (biomarkers), cell proliferation and cytotoxicity were evaluated to identify activities of 14-3-3η relative to vehicle control. The resultant 14-3-3η BioMAP profile was analyzed and compared in a similarity search with more than 3000 compounds in the BioMAP database to identify common mechanistic signatures using Pearson’s correlation.
Results 14-3-3η messenger RNA (mRNA) is highly expressed in all BioMAP Diversity Plus Systems with stimulus coupled increases detected in B cell and Fibroblast based systems. The activity profile for 14-3-3η in BioMAP shows highly selective effects in two systems: 1. HPNo, a non-stimulated vascular endothelial cell-PBMC co-culture; 2. BT, a stimulated co-culture of CD19+ B cells plus PBMC modeling T-cell dependent B cell activation. 14-3-3η caused an increase in levels of VCAM-1 and TNFα in HPNo and sIL-6 production in BT, activities consistent with an inflammatory phenotype. Comparison of the profile to the BioMAP database identified mechanistic matches with several pro-inflammatory TLR-like agonists including Pam3CSK4, a TLR-2/-1 agonist (r = 0.735), Flagellin, a TLR-5 agonist (r = 0.723) and HKLM, a TLR-2 agonist (r = 0.721). Of interest, the BioMAP profile of 14-3-3η was not similar to LPS, a potential bacterial endotoxin contaminant of biological preparations that also has TLR-agonist activities.
Conclusion The BioMAP profile for 14-3-3η is consistent with activation of B cell responses that correlates with pro-inflammatory activity associated with disease-relevant biomarkers. This data supports the hypothesis that 14-3-3η may serve both as a diagnostic marker in early RA as well as an important target for therapeutic intervention.5
References: 1. Arthritis Res Ther. 2014;16(2). 2. J Rheumatol. 2007 Aug;34(8): 1650-7. 3. Drug Discov Today. 2014;19:113-25. 4. J Biomol Screen. 2013;18:1260-9. 5. Semin Cell Dev Biol. 2011; 22(7):705-12.
E. L. Berg,
W. P. Maksymowych,
Augurex Life Sciences Corp,
Augurex Life Sciences Corp,
Augurex Life Sciences Corp.,
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ACR Meeting Abstracts - http://acrabstracts.org/abstract/profiling-14-3-3%ce%b7-in-human-primary-cell-based-biomap-disease-models-reveals-a-unique-pro-inflammatory-phenotypic-signature-consistent-with-ra-inflammation-biology/