Bhingarkar, Aboli
(2022)
Duvelisib is a novel NFAT inhibitor that attenuates adalimumab immunogenicity.
Master's Thesis, University of Pittsburgh.
(Unpublished)
Abstract
The immunogenicity of biologics is a major problem in rheumatoid arthritis (RA) because it neutralizes the activity of biologics such as adalimumab via the development of anti-drug antibodies (ADA) that can lead to disease relapse. Although the current prevention strategy involving co-administration of methotrexate (MTX) with the biologic is effective at suppressing ADA development, patients often terminate MTX therapy because of the toxicities associated with its use. Alternative treatment strategies that are well-tolerated and as effective as MTX are required. A recent GWAS found that Nuclear Factor of Activated T cells 2 (NFATC2) was associated with a higher risk of immunotoxicity. Thus, targeting the NFAT pathway may mitigate the risk of drug-induced immunogenicity and protect against disease relapse. Several pharmacological inhibitors are available, but they come with their own set of limitations making their use not feasible. Hence, there is a need to identify novel NFAT inhibitors. Our team performed computational analysis which revealed five compounds potentially binding to NFATC2. We screened the novel compounds using Jurkat-Lucia NFAT cells to measure NFAT inhibitor activity. In the current study, we focus on one of the compounds, duvelisib, that exhibited binding to the DNA pocket of NFATC2 by the computational analysis and strongly attenuated NFAT attenuation at a concentration of 10uM in our inhibitor screen. Through a series of experiments investigating the mechanism behind NFAT inhibition, we demonstrate that duvelisib potentially binds to NFATC2/C1 and does not affect calcium mobilization, calcineurin activation by calmodulin, the phosphatase activity of calcineurin, or nuclear translocation of NFATC2/C1. Interestingly, we also show that treatment with the GSK3β inhibitor, tideglusib abrogated NFAT inhibition demonstrated by other PI3Kγδ inhibitors, but not for duvelisib. This suggests that duvelisib-mediated NFAT suppression was independent of its PI3K suppressive effects. We tested duvelisib in our mice model of adalimumab-induced immunogenicity found that duvelisib protected mice from the development of ADA. Furthermore, we demonstrate that duvelisib suppresses T cell proliferation and B cell antibody secretion in murine and human cells. Our study features duvelisib as a novel NFAT inhibitor that is effective in mitigating drug-induced immunogenicity. Our future directions involve strengthening the mechanistic evidence by using molecular biology, genetic, and mutational approaches.
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Details
| Item Type: |
University of Pittsburgh ETD
|
| Status: |
Unpublished |
| Creators/Authors: |
|
| ETD Committee: |
|
| Date: |
9 September 2022 |
| Date Type: |
Publication |
| Defense Date: |
4 August 2022 |
| Approval Date: |
9 September 2022 |
| Submission Date: |
30 August 2022 |
| Access Restriction: |
2 year -- Restrict access to University of Pittsburgh for a period of 2 years. |
| Number of Pages: |
57 |
| Institution: |
University of Pittsburgh |
| Schools and Programs: |
School of Pharmacy > Pharmaceutical Sciences |
| Degree: |
MS - Master of Science |
| Thesis Type: |
Master's Thesis |
| Refereed: |
Yes |
| Uncontrolled Keywords: |
Drug-mediated immunogenicity, anti-drug antibodies, Nuclear Factor of Activated T cells (NFAT) |
| Date Deposited: |
09 Sep 2022 15:24 |
| Last Modified: |
09 Sep 2022 15:24 |
| URI: |
http://d-scholarship.pitt.edu/id/eprint/43684 |
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