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Oral cancer treatment by targeted drug delivery system with an anti-desmoglein monoclonal antibody
Abstracts from the 41st Annual Meeting / Journal of Dermatological Science 86 (2017) e1–e95
P04-03[I-4] Oral cancer treatment by targeted drug delivery system with an anti-desmoglein monoclonal antibody Michiyoshi Kouno 1,∗ , Masaki Minabe 2 , Tetsuhiko Tachikawa 3 , John R. Stanley 4 1
Department of Dermatology, Tokyo Dental College Ichikawa General Hospital, Chiba, Japan 2 Department of Oral Medicine, Tokyo Dental College Ichikawa General Hospital, Chiba, Japan 3 Division of Molecular Diagnosis and Cancer Prevention, Saitama Cancer Center, Saitama, Japan 4 Department of Dermatology, University of Pennsylvania, Philadelphia, PA, United States Desmoglein (Dsg) 3 is overexpressed in oral squamous cell carcinoma (OSCC). We hypothesized that Dsg3 could be used for targeted therapy and developed a method of targeting an antitumor agent, tumor necrosis factor-related apoptosis-inducing ligand (TRAIL), to the OSCC by fusing it with Px44, a non-pathogenic antiDsg3 single chain variable fragment (scFv) antibody cloned from a pemphigus patient. TRAIL has been shown to cause apoptosis of rapidly proliferating tumor cells, and we previously showed that Px44TRAIL had stable proapoptotic activity on Jurkat lymphoma cells even after incubation for 48 h at 37 ◦ C and that it did not have toxic effects on normal skin. In this study, we tested the activity of Px44TRAIL on human OSCC cell lines derived from primary tumor and metastatic lymph nodes of 6 OSCC patients. Real time qPCR showed that 11 of the 12 cell lines expressed Dsg3 mRNA. Cells from the metastatic lymph nodes of each patient expressed more Dsg3 than those from the primary tumor of the same patient. Immunofluorescence (IF) detected Px 44TRAIL binding on the cell surface of the 11 cell lines expressing Dsg3, but not on the Dsg3-negative line. TRAIL alone caused apoptosis in 6 of these lines over 16 h, whereas Px44TRAIL targeted for only 2 h (then washed) caused apoptosis of up to 50% of cells in 9 of the 12 cell lines at 16 h. AM3-13TRAIL (in which AM3-13 is an irrelevant scFv) did not bind on the cells or cause apoptosis. Finally, when injected into human SCC organ culture, IF detected Px44TRAIL binding on the invading tumor cells. Our data show the feasibility of targeting TRAIL to SCC, and suggest such therapy may be useful for oral cancer treatment. http://dx.doi.org/10.1016/j.jdermsci.2017.02.078 P04-04[C06-1] Efficacy of the injection of botulinum toxin B on Raynaud’s phenomenon in patients with systemic sclerosis: Single-blind, randomized trial
Sei-ichiro Motegi 1,∗ , Kazuya Yamada 1 , Akihito Uehara 1 , Akiko Sekiguchi 1 , Yuki Date 2 , Tetsuya Nakamura 2 , Osamu Ishikawa 1 1 Department of Dermatology, Gunma University Graduate School of Medicine, Maebashi, Japan 2 Clinical Investigation and Research Unit, Gunma University Graduate School of Medicine, Maebashi, Japan Patients with systemic scleroderma (SSc) typically develop Raynaud’s phenomenon (RP) and persistent digital ischemia and often develop digital ulcers (DUs). Currently, there is no satisfactory
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treatment for RP in SSc. We recently demonstrated that botulinum toxin A (BTX-A) injection was effective for the treatment of RP in SSc patients. However, the efficacy of BTX-B injection has never been examined. The objective of the present study was to assess the efficacy and safety of BTX-B on RP in SSc patients. In the prospective, single-blind, randomized trial, 45 SSc patients with RP were blinded and randomly divided into 4 groups; no treatment control group and treatment groups with 250, 1000 or 2000 U of BTX-B injections per hand. In 4 weeks after injection, the pain VAS and Raynaud’s score (indicating the severity of RP in SSc patients) in the 1000- and 2000-U-treated groups were significantly lower than those in the control group and 250-U-treated group. The beneficial effects of 1000 and 2000 U treatment continued until 16 weeks after one-time injection. The skin temperature recovery after cold-water stimulation in the 2000-U-treated group at 4 weeks after injection was significantly improved compared to those in the control group and 250-U-treated group. The number of DUs in the 1000- and 2000-U-treated groups were significantly lower than in the control group. Transient muscle weakness occurred in 1 patient in the 2000-U-treated group, but no systemic or local adverse events were observed in any patients. We conclude that 1000 and 2000 U BTXB injection significantly improved the activity of RP in SSc patients without any serious adverse events, suggesting that BTX-B might have potential as a long-term preventive and therapeutic agent for RP and DUs in SSc patients. http://dx.doi.org/10.1016/j.jdermsci.2017.02.079 P04-05[C11-1] Activated regulatory T cells are increased in patients with alopecia areata for suppressing disease acitivity Ryoji Kubo ∗ , Shinnosuke Muramatsu, Yoko Sagawa, Chiyo Saito, Saori Kasuya, Akiko Nishioka, Emi Nishida, Akimichi Morita Department of Geriatric and Environmental Dermatology, Nagoya City University Graduate School of Medical Sciences, Nagoya, Japan Alopecia areata is one of refractory inflammatory skin disorders. However the precise mechanism remains obscure and appropriate treatment for severe cases should be developed even patients experience self-regression. We previously evaluated the number of circulating CD4+ CD 25+ regulatory T cells were increased substantially, but not statistically significant in alopecia patients compared with healthy controls (4.69 ± 1.24% vs. 3.81 ± 1.12%, p = 0.074). In the present study, we examined three distinct Treg subsets: activated Treg (aTreg), resting Treg (rTreg), and non-suppressive T cells (non-Treg). aTreg have the strongest suppressive activity among the three subsets and rTreg are moderately suppressive. We examined these subsets from the peripheral blood in 12 alopecia areata and 7 atopic dermatitis patients. aTreg levels and aTreg + rTreg levels of alopecia areata were significantly higher than those of healthy controls (n = 11) (aTreg: 1.87 ± 1.27% vs. 0.92 ± 0.44%, p = 0.022; aTreg + rTreg: 3.22 ± 1.55% vs. 2.38 ± 0.44%, p = 0.045). In the fourgroup comparison, alopecia areata, atopic dermatitis, psoriasis (n = 15) and healthy controls, aTreg + rTreg levels in alopecia areata tended to be higher among these groups (no significance). aTreg levels in patients not having alopecia total is were significantly higher than that of healthy controls (2.02 ± 0.95% vs. 0.92 ± 0.44%, p = 0.047). aTreg and rTreg levels were negatively correlated with the disease duration (aTreg: r = −0.306; aTreg + rTreg: r = −0.544). These results indicated that increased aTreg + rTreg would suppress the disease activity at early phase and induce spontaneous remission to some extent in alopecia areata patients. Therefore, the
P04-03[I-4] Oral cancer treatment by targeted drug delivery system with an anti-desmoglein monoclonal antibody Michiyoshi Kouno 1,∗ , Masaki Minabe 2 , Tetsuhiko Tachikawa 3 , John R. Stanley 4 1
Department of Dermatology, Tokyo Dental College Ichikawa General Hospital, Chiba, Japan 2 Department of Oral Medicine, Tokyo Dental College Ichikawa General Hospital, Chiba, Japan 3 Division of Molecular Diagnosis and Cancer Prevention, Saitama Cancer Center, Saitama, Japan 4 Department of Dermatology, University of Pennsylvania, Philadelphia, PA, United States Desmoglein (Dsg) 3 is overexpressed in oral squamous cell carcinoma (OSCC). We hypothesized that Dsg3 could be used for targeted therapy and developed a method of targeting an antitumor agent, tumor necrosis factor-related apoptosis-inducing ligand (TRAIL), to the OSCC by fusing it with Px44, a non-pathogenic antiDsg3 single chain variable fragment (scFv) antibody cloned from a pemphigus patient. TRAIL has been shown to cause apoptosis of rapidly proliferating tumor cells, and we previously showed that Px44TRAIL had stable proapoptotic activity on Jurkat lymphoma cells even after incubation for 48 h at 37 ◦ C and that it did not have toxic effects on normal skin. In this study, we tested the activity of Px44TRAIL on human OSCC cell lines derived from primary tumor and metastatic lymph nodes of 6 OSCC patients. Real time qPCR showed that 11 of the 12 cell lines expressed Dsg3 mRNA. Cells from the metastatic lymph nodes of each patient expressed more Dsg3 than those from the primary tumor of the same patient. Immunofluorescence (IF) detected Px 44TRAIL binding on the cell surface of the 11 cell lines expressing Dsg3, but not on the Dsg3-negative line. TRAIL alone caused apoptosis in 6 of these lines over 16 h, whereas Px44TRAIL targeted for only 2 h (then washed) caused apoptosis of up to 50% of cells in 9 of the 12 cell lines at 16 h. AM3-13TRAIL (in which AM3-13 is an irrelevant scFv) did not bind on the cells or cause apoptosis. Finally, when injected into human SCC organ culture, IF detected Px44TRAIL binding on the invading tumor cells. Our data show the feasibility of targeting TRAIL to SCC, and suggest such therapy may be useful for oral cancer treatment. http://dx.doi.org/10.1016/j.jdermsci.2017.02.078 P04-04[C06-1] Efficacy of the injection of botulinum toxin B on Raynaud’s phenomenon in patients with systemic sclerosis: Single-blind, randomized trial
Sei-ichiro Motegi 1,∗ , Kazuya Yamada 1 , Akihito Uehara 1 , Akiko Sekiguchi 1 , Yuki Date 2 , Tetsuya Nakamura 2 , Osamu Ishikawa 1 1 Department of Dermatology, Gunma University Graduate School of Medicine, Maebashi, Japan 2 Clinical Investigation and Research Unit, Gunma University Graduate School of Medicine, Maebashi, Japan Patients with systemic scleroderma (SSc) typically develop Raynaud’s phenomenon (RP) and persistent digital ischemia and often develop digital ulcers (DUs). Currently, there is no satisfactory
e27
treatment for RP in SSc. We recently demonstrated that botulinum toxin A (BTX-A) injection was effective for the treatment of RP in SSc patients. However, the efficacy of BTX-B injection has never been examined. The objective of the present study was to assess the efficacy and safety of BTX-B on RP in SSc patients. In the prospective, single-blind, randomized trial, 45 SSc patients with RP were blinded and randomly divided into 4 groups; no treatment control group and treatment groups with 250, 1000 or 2000 U of BTX-B injections per hand. In 4 weeks after injection, the pain VAS and Raynaud’s score (indicating the severity of RP in SSc patients) in the 1000- and 2000-U-treated groups were significantly lower than those in the control group and 250-U-treated group. The beneficial effects of 1000 and 2000 U treatment continued until 16 weeks after one-time injection. The skin temperature recovery after cold-water stimulation in the 2000-U-treated group at 4 weeks after injection was significantly improved compared to those in the control group and 250-U-treated group. The number of DUs in the 1000- and 2000-U-treated groups were significantly lower than in the control group. Transient muscle weakness occurred in 1 patient in the 2000-U-treated group, but no systemic or local adverse events were observed in any patients. We conclude that 1000 and 2000 U BTXB injection significantly improved the activity of RP in SSc patients without any serious adverse events, suggesting that BTX-B might have potential as a long-term preventive and therapeutic agent for RP and DUs in SSc patients. http://dx.doi.org/10.1016/j.jdermsci.2017.02.079 P04-05[C11-1] Activated regulatory T cells are increased in patients with alopecia areata for suppressing disease acitivity Ryoji Kubo ∗ , Shinnosuke Muramatsu, Yoko Sagawa, Chiyo Saito, Saori Kasuya, Akiko Nishioka, Emi Nishida, Akimichi Morita Department of Geriatric and Environmental Dermatology, Nagoya City University Graduate School of Medical Sciences, Nagoya, Japan Alopecia areata is one of refractory inflammatory skin disorders. However the precise mechanism remains obscure and appropriate treatment for severe cases should be developed even patients experience self-regression. We previously evaluated the number of circulating CD4+ CD 25+ regulatory T cells were increased substantially, but not statistically significant in alopecia patients compared with healthy controls (4.69 ± 1.24% vs. 3.81 ± 1.12%, p = 0.074). In the present study, we examined three distinct Treg subsets: activated Treg (aTreg), resting Treg (rTreg), and non-suppressive T cells (non-Treg). aTreg have the strongest suppressive activity among the three subsets and rTreg are moderately suppressive. We examined these subsets from the peripheral blood in 12 alopecia areata and 7 atopic dermatitis patients. aTreg levels and aTreg + rTreg levels of alopecia areata were significantly higher than those of healthy controls (n = 11) (aTreg: 1.87 ± 1.27% vs. 0.92 ± 0.44%, p = 0.022; aTreg + rTreg: 3.22 ± 1.55% vs. 2.38 ± 0.44%, p = 0.045). In the fourgroup comparison, alopecia areata, atopic dermatitis, psoriasis (n = 15) and healthy controls, aTreg + rTreg levels in alopecia areata tended to be higher among these groups (no significance). aTreg levels in patients not having alopecia total is were significantly higher than that of healthy controls (2.02 ± 0.95% vs. 0.92 ± 0.44%, p = 0.047). aTreg and rTreg levels were negatively correlated with the disease duration (aTreg: r = −0.306; aTreg + rTreg: r = −0.544). These results indicated that increased aTreg + rTreg would suppress the disease activity at early phase and induce spontaneous remission to some extent in alopecia areata patients. Therefore, the