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4. Additional file 1: of Methotrexate upregulates circadian transcriptional factors PAR bZIP to induce apoptosis on rheumatoid arthritis synovial fibroblasts

Author

Kohjin Suzuki, Yoshida, Kohsuke, Ueha, Takeshi, Kaneshiro, Kenta, Nakai, Ayako, Naonori Hashimoto, Koto Uchida, Hashimoto, Teppei, Kawasaki, Yoshiko, Shibanuma, Nao, Nakagawa, Natsuko, Yoshitada Sakai, and Hashiramoto, Akira

Subjects
endocrine system
Abstract

The interactions of circadian clock genes and its relative factors. BMAL/CLOCK and PER/CRY create the circadian rhythm via E-box, and DBP, TEF, HLF, E4BP4 via D-box. BMAL and CLOCK heterodimerize and bind to E-box elements on promoter regions of Per and Cry genes to induce their transcription. Thereafter, PER and CRY proteins heterodimerize to inhibit activities of their own and other E-box regulated promoters. (PDF 214 kb)

Published
2018
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5. Additional file 2: of Methotrexate upregulates circadian transcriptional factors PAR bZIP to induce apoptosis on rheumatoid arthritis synovial fibroblasts

Author

Kohjin Suzuki, Yoshida, Kohsuke, Ueha, Takeshi, Kaneshiro, Kenta, Nakai, Ayako, Naonori Hashimoto, Koto Uchida, Hashimoto, Teppei, Kawasaki, Yoshiko, Shibanuma, Nao, Nakagawa, Natsuko, Yoshitada Sakai, and Hashiramoto, Akira

Subjects
musculoskeletal diseases, immune system diseases, skin and connective tissue diseases
Abstract

Detailed information of patients with RA enrolled in this study. Joint samples obtained from 10 different patients to establish primary cultured synovial fibroblast cell lines. Age, sex, disease duration, CRP, DAS28-ESR, and treatment with MTX, PSL, and other DMARDs shown in the table. (PDF 252 kb)

Published
2018
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6. Additional file 5: of Methotrexate upregulates circadian transcriptional factors PAR bZIP to induce apoptosis on rheumatoid arthritis synovial fibroblasts

Author

Kohjin Suzuki, Yoshida, Kohsuke, Ueha, Takeshi, Kaneshiro, Kenta, Nakai, Ayako, Naonori Hashimoto, Koto Uchida, Hashimoto, Teppei, Kawasaki, Yoshiko, Shibanuma, Nao, Nakagawa, Natsuko, Yoshitada Sakai, and Hashiramoto, Akira

Subjects
musculoskeletal diseases
Abstract

The mRNA expression of circadian clock genes over time. mRNA expression of circadian clock genes measured at â 4 h (before synchronization), 0 h (just before MTX stimulation), 24 h, 32 h, and 48 h. Controls and 10/100 nM of MTX showed almost the same expression rhythms, and MTX influenced their expression levels. (PDF 264 kb)

Published
2018
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7. Additional file 6: of Methotrexate upregulates circadian transcriptional factors PAR bZIP to induce apoptosis on rheumatoid arthritis synovial fibroblasts

Author

Kohjin Suzuki, Yoshida, Kohsuke, Ueha, Takeshi, Kaneshiro, Kenta, Nakai, Ayako, Naonori Hashimoto, Koto Uchida, Hashimoto, Teppei, Kawasaki, Yoshiko, Shibanuma, Nao, Nakagawa, Natsuko, Yoshitada Sakai, and Hashiramoto, Akira

Subjects
endocrine system
Abstract

Two different transcriptional pathways by which MTX induces apoptosis to synovial fibroblasts: PAR bZIPâ Per2 transcriptional pathway and PAR bZIPâ Bik transcriptional pathway. We propose MTX induces apoptosis in synovial cells through activated binding of PAR bZIP to D-box in two different genes, Per2 and Bik, and these dual pathways work independently but synergistically. (PDF 206 kb)

Published
2018
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8. Additional file 3: of Methotrexate upregulates circadian transcriptional factors PAR bZIP to induce apoptosis on rheumatoid arthritis synovial fibroblasts

Author

Kohjin Suzuki, Yoshida, Kohsuke, Ueha, Takeshi, Kaneshiro, Kenta, Nakai, Ayako, Naonori Hashimoto, Koto Uchida, Hashimoto, Teppei, Kawasaki, Yoshiko, Shibanuma, Nao, Nakagawa, Natsuko, Yoshitada Sakai, and Hashiramoto, Akira

Abstract

Plasmid constructs of Per2/Bik promoter. D-box(+), plasmid constructs containing D-box. D-box(−), plasmid constructs without D-box. D-box motifs of Per2 promoter were mutated from 5′-TTATGTAA-3′ to 5′-CGCCAGGC-3′ (−372 to −365), and 5′-TTACGTAA-3′ to 5′-CAGCGTAA-3′ (−47 to −40). Human Bik promoter containing D-box (−780 to +176) and human Bik promoter without D-box (−260 to +323) constructed. (PDF 215 kb)

Published
2018
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9. Additional file 4: of Methotrexate upregulates circadian transcriptional factors PAR bZIP to induce apoptosis on rheumatoid arthritis synovial fibroblasts

Author

Kohjin Suzuki, Yoshida, Kohsuke, Ueha, Takeshi, Kaneshiro, Kenta, Nakai, Ayako, Naonori Hashimoto, Koto Uchida, Hashimoto, Teppei, Kawasaki, Yoshiko, Shibanuma, Nao, Nakagawa, Natsuko, Yoshitada Sakai, and Hashiramoto, Akira

Subjects
musculoskeletal diseases, immune system diseases, heterocyclic compounds, skin and connective tissue diseases
Abstract

Cell viability of MTX-treated fibroblasts. Cell viability of RA synovial fibroblasts measured by WST-8 assay after 24 h of stimulation of MTX (1 pM to 1 ÎźM). MTX (1 and 10 nM) significantly decreased cell viability as shown in Fig. 1, while 1â 100 pM of MTX did not. (PDF 274 kb)

Published
2018
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14. Circulating cell free DNA: a marker to predict the therapeutic response for biological DMARDs in rheumatoid arthritis.

Author

Hashimoto, Teppei, Yoshida, Kohsuke, Hashimoto, Naonori, Nakai, Ayako, Kaneshiro, Kenta, Suzuki, Kohjin, Kawasaki, Yoshiko, Shibanuma, Nao, and Hashiramoto, Akira

Subjects
DNA, RHEUMATOID arthritis, POLYMERASE chain reaction, RHEUMATISM, BLOOD sedimentation
Abstract

Aim To evaluate the correlation between circulating cell-free DNA (ccfDNA) in plasma and clinical disease activities in patients with rheumatoid arthritis (RA). Method The study group included 30 patients with RA who started biological disease-modifying anti-rheumatic drugs (DMARDs) therapy. The concentration of ccfDNA in plasma was measured by quantitative real-time polymerase chain reaction at baseline to 24 weeks in every 4-week period from 30 patients and 21 healthy individuals. We also evaluated the correlation between ccfDNA and the clinical activity or the therapeutic response for biological DMARDs, using the simplified disease activity index (SDAI), Disease Activity Score of 28 joints (erythrocyte sedimentation rate) and the European League Against Rheumatism (EULAR) response criteria. Synovial fluid samples of knee joints were collected from 13 patients with RA and 12 with osteoarthritis (OA) to measure ccfDNA. Result The concentration of ccfDNA in RA patients at baseline was higher than healthy controls ( P = 0.016). After introducing biological DMARDs, ccfDNA was increased until 8 weeks from the baseline, and decreased after 12 weeks. The average of SDAI was improved in all patients enrolled. At 12 weeks after treatment, 15 patients were good responders to the EULAR response criteria, nine showed moderate response and six showed no response. ccfDNA in good responders was increased until 8 weeks, while those of moderate or no response were not ( P = 0.042). In joint fluid of RA patients, ccfDNA was remarkably increased as compared to those from OA ( P = 0.00011). Conclusion After introducing biological DMARDs, increase of ccfDNA at 8 weeks was associated with improvement of disease activities. Compared with biomarkers reported, ccfDNA is able to predict the early therapeutic effects of biological DMARDs in RA patients. [ABSTRACT FROM AUTHOR]

Published
2017
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