Caspase‐8 Variant G Regulates Rheumatoid Arthritis Fibroblast‐Like Synoviocyte Aggressive Behavior
Objective Fibroblast‐like synoviocytes (FLS) play a pivotal role in rheumatoid arthritis (RA) by contributing to synovial inflammation and progressive joint damage. An imprinted epigenetic state is associated with the FLS aggressive phenotype. We identified CASP8 (encoding for caspase‐8) as a differ...
| Main Authors: | , , , , , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Wiley
2022-04-01
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| Series: | ACR Open Rheumatology |
| Online Access: | https://doi.org/10.1002/acr2.11384 |
| _version_ | 1818365070892072960 |
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| author | Cecilia Ansalone Richard I. Ainsworth Gyrid Nygaard Rizi Ai Edward B. Prideaux Deepa Hammaker Narayanan B. Perumal Ken Weichert Frances Tung Lalitha Kodandapani J. Michael Sauder Elisabeth C. Mertsching Robert J. Benschop David L. Boyle Wei Wang Gary S. Firestein |
| author_facet | Cecilia Ansalone Richard I. Ainsworth Gyrid Nygaard Rizi Ai Edward B. Prideaux Deepa Hammaker Narayanan B. Perumal Ken Weichert Frances Tung Lalitha Kodandapani J. Michael Sauder Elisabeth C. Mertsching Robert J. Benschop David L. Boyle Wei Wang Gary S. Firestein |
| author_sort | Cecilia Ansalone |
| collection | DOAJ |
| description | Objective Fibroblast‐like synoviocytes (FLS) play a pivotal role in rheumatoid arthritis (RA) by contributing to synovial inflammation and progressive joint damage. An imprinted epigenetic state is associated with the FLS aggressive phenotype. We identified CASP8 (encoding for caspase‐8) as a differentially marked gene and evaluated its pathogenic role in RA FLSs. Methods RA FLS lines were obtained from synovial tissues at arthroplasty and used at passage 5‐8. Caspase‐8 was silenced using small interfering RNA, and its effect was determined in cell adhesion, migration and invasion assays. Quantitative reverse transcription PCR and western blot were used to assess gene and protein expression, respectively. A caspase‐8 selective inhibitor was used determine the role of enzymatic activity on FLS migration and invasion. Caspase‐8 isoform transcripts and epigenetic marks in FLSs were analyzed in FLS public databases. Crystal structures of caspase‐8B and G were determined. Results Caspase‐8 deficiency in RA FLSs reduced cell adhesion, migration, and invasion independent of its catalytic activity. Epigenetic and transcriptomic analyses of RA FLSs revealed that a specific caspase‐8 isoform, variant G, is the dominant isoform expressed (~80% of total caspase‐8) and induced by PDGF. The crystal structures of caspase‐8 variant G and B were identical except for a unique unstructured 59 amino acid N‐terminal domain in variant G. Selective knockdown of caspase‐8G was solely responsible for the effects of caspase‐8 on calpain activity and cell invasion in FLS. Conclusion Blocking caspase‐8 variant G could decrease cell invasion in diseases like RA without the potential deleterious effects of nonspecific caspase‐8 inhibition. |
| first_indexed | 2024-12-13T22:14:25Z |
| format | Article |
| id | doaj.art-7bf9364f9ed048b68c6e9061d4e747da |
| institution | Directory Open Access Journal |
| issn | 2578-5745 |
| language | English |
| last_indexed | 2024-12-13T22:14:25Z |
| publishDate | 2022-04-01 |
| publisher | Wiley |
| record_format | Article |
| series | ACR Open Rheumatology |
| spelling | doaj.art-7bf9364f9ed048b68c6e9061d4e747da2022-12-21T23:29:38ZengWileyACR Open Rheumatology2578-57452022-04-014428829910.1002/acr2.11384Caspase‐8 Variant G Regulates Rheumatoid Arthritis Fibroblast‐Like Synoviocyte Aggressive BehaviorCecilia Ansalone0Richard I. Ainsworth1Gyrid Nygaard2Rizi Ai3Edward B. Prideaux4Deepa Hammaker5Narayanan B. Perumal6Ken Weichert7Frances Tung8Lalitha Kodandapani9J. Michael Sauder10Elisabeth C. Mertsching11Robert J. Benschop12David L. Boyle13Wei Wang14Gary S. Firestein15University of California San Diego La Jolla CaliforniaUniversity of California San Diego La Jolla CaliforniaUniversity of California San Diego La Jolla CaliforniaUniversity of California San Diego La Jolla CaliforniaUniversity of California San Diego La Jolla CaliforniaUniversity of California San Diego La Jolla CaliforniaLilly Biotechnology Center San Diego CaliforniaLilly Biotechnology Center San Diego CaliforniaLilly Biotechnology Center San Diego CaliforniaLilly Biotechnology Center San Diego CaliforniaLilly Biotechnology Center San Diego CaliforniaLilly Biotechnology Center San Diego CaliforniaEli Lilly and Company Indianapolis IndianaUniversity of California San Diego La Jolla CaliforniaUniversity of California San Diego La Jolla CaliforniaUniversity of California San Diego La Jolla CaliforniaObjective Fibroblast‐like synoviocytes (FLS) play a pivotal role in rheumatoid arthritis (RA) by contributing to synovial inflammation and progressive joint damage. An imprinted epigenetic state is associated with the FLS aggressive phenotype. We identified CASP8 (encoding for caspase‐8) as a differentially marked gene and evaluated its pathogenic role in RA FLSs. Methods RA FLS lines were obtained from synovial tissues at arthroplasty and used at passage 5‐8. Caspase‐8 was silenced using small interfering RNA, and its effect was determined in cell adhesion, migration and invasion assays. Quantitative reverse transcription PCR and western blot were used to assess gene and protein expression, respectively. A caspase‐8 selective inhibitor was used determine the role of enzymatic activity on FLS migration and invasion. Caspase‐8 isoform transcripts and epigenetic marks in FLSs were analyzed in FLS public databases. Crystal structures of caspase‐8B and G were determined. Results Caspase‐8 deficiency in RA FLSs reduced cell adhesion, migration, and invasion independent of its catalytic activity. Epigenetic and transcriptomic analyses of RA FLSs revealed that a specific caspase‐8 isoform, variant G, is the dominant isoform expressed (~80% of total caspase‐8) and induced by PDGF. The crystal structures of caspase‐8 variant G and B were identical except for a unique unstructured 59 amino acid N‐terminal domain in variant G. Selective knockdown of caspase‐8G was solely responsible for the effects of caspase‐8 on calpain activity and cell invasion in FLS. Conclusion Blocking caspase‐8 variant G could decrease cell invasion in diseases like RA without the potential deleterious effects of nonspecific caspase‐8 inhibition.https://doi.org/10.1002/acr2.11384 |
| spellingShingle | Cecilia Ansalone Richard I. Ainsworth Gyrid Nygaard Rizi Ai Edward B. Prideaux Deepa Hammaker Narayanan B. Perumal Ken Weichert Frances Tung Lalitha Kodandapani J. Michael Sauder Elisabeth C. Mertsching Robert J. Benschop David L. Boyle Wei Wang Gary S. Firestein Caspase‐8 Variant G Regulates Rheumatoid Arthritis Fibroblast‐Like Synoviocyte Aggressive Behavior ACR Open Rheumatology |
| title | Caspase‐8 Variant G Regulates Rheumatoid Arthritis Fibroblast‐Like Synoviocyte Aggressive Behavior |
| title_full | Caspase‐8 Variant G Regulates Rheumatoid Arthritis Fibroblast‐Like Synoviocyte Aggressive Behavior |
| title_fullStr | Caspase‐8 Variant G Regulates Rheumatoid Arthritis Fibroblast‐Like Synoviocyte Aggressive Behavior |
| title_full_unstemmed | Caspase‐8 Variant G Regulates Rheumatoid Arthritis Fibroblast‐Like Synoviocyte Aggressive Behavior |
| title_short | Caspase‐8 Variant G Regulates Rheumatoid Arthritis Fibroblast‐Like Synoviocyte Aggressive Behavior |
| title_sort | caspase 8 variant g regulates rheumatoid arthritis fibroblast like synoviocyte aggressive behavior |
| url | https://doi.org/10.1002/acr2.11384 |
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