Molecular Therapy - Nucleic Acids

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Molecular Therapy - Nucleic Acids A Chymase Inhibitory RNA Aptamer Improves Cardiac Function and Survival after Myocardial Infarction Denan Jin, Shinji Takai, Yosuke Nonaka, Satoko Yamazaki, Masatoshi Fujiwara, Yoshikazu Nakamura Molecular Therapy - Nucleic Acids Volume 14, Pages 41-51 (March 2019) DOI: 10.1016/j.omtn.2018.11.001 Copyright © 2018 The Authors Terms and Conditions

Figure 1 Characteristics of HA28 (A) Structure of HA28. (B) Inhibitory efficacy of HA28 on purified hamster chymase. Each value is the mean of two measurements in the same concentration. (C) Blood concentration 24 hr after final subcutaneous injection of HA28 in the hamster MI model. Each group contained 8 hamsters. HA28 treatment was started 1 day before the induction of MI and continued for 3 days. Molecular Therapy - Nucleic Acids 2019 14, 41-51DOI: (10.1016/j.omtn.2018.11.001) Copyright © 2018 The Authors Terms and Conditions

Figure 2 Cardiac Functional and Histological Examinations (A) Representative M-mode echocardiograms and Doppler spectra of mitral inflow in the sham-operated group (sham) and MI hamsters treated with vehicle (MI) or HA28 3 days after the operation. (B) Effects of HA28 on the LV ejection fraction (EF) and the ratio of the early (E) to late (A) ventricular filling velocities (E:A). HA28 at a dosage of 1, 3, or 10 mg/kg was subcutaneously injected 1 day before surgery and continued for 3 days thereafter. (C) Representative azan Mallory staining, toluidine blue staining, and chymase immunohistochemical staining of cardiac sections obtained 3 days after surgery. (D) Effects of HA28 on chymase-positive mast cells and cardiac chymase activities 3 days after MI. HA28 treatment was started 1 day before the induction of MI and continued for 3 days. Dd, left ventricular dimension end diastole; Ds, left ventricular dimension end systole; E, E-wave velocity; A, A-wave velocity. **p < 0.01 versus sham; #p < 0.05, ##p < 0.01 versus vehicle. Each group contained 6–8 animals. Significant differences among the mean values of multiple groups were evaluated with one-way ANOVA followed by post hoc analysis (Fisher’s test). Molecular Therapy - Nucleic Acids 2019 14, 41-51DOI: (10.1016/j.omtn.2018.11.001) Copyright © 2018 The Authors Terms and Conditions

Figure 3 Cumulative Mortality Rates of MI Hamsters Treated with Vehicle (n = 21) or HA28 (n = 24) HA28 at the dosage of 1 mg/kg was subcutaneously injected 1 day after LCA ligation once a day and continued for 14 days. The survival curves of individual groups were compared with the log-rank test. Molecular Therapy - Nucleic Acids 2019 14, 41-51DOI: (10.1016/j.omtn.2018.11.001) Copyright © 2018 The Authors Terms and Conditions

Figure 4 Effect of HA28 on Cardiac Activity, Infarct Size, and Ratio of Heart Weight to Body Weight 14 Days after Surgery HA28 at the dosage of 1 mg/kg was subcutaneously injected 1 day after LCA ligation once a day and continued for 14 days. (A) Representative azan Mallory staining from the sham-operated group and MI groups treated with vehicle (MI) or HA28. (B) Ratio of the whole-heart weight to body weight, infarct size, and cardiac chymase activity in the sham-operated group (n = 8) and MI hamsters treated with vehicle (V, n = 14) or HA28 (n = 22). **p < 0.01, ***p < 0.001 versus sham; ###p < 0.001 versus vehicle. Significant differences between the mean values of two groups were evaluated with the Student’s t test for unpaired data. Significant differences among the mean values of multiple groups were evaluated with one-way ANOVA followed by post hoc analysis (Fisher’s test). Molecular Therapy - Nucleic Acids 2019 14, 41-51DOI: (10.1016/j.omtn.2018.11.001) Copyright © 2018 The Authors Terms and Conditions

Figure 5 Relative Cardiac mRNA Levels of Chymase, TGF-β1, MMP9, MMP2, and Collagen I in the Sham-Operated Group (Sham) and MI Groups Treated with Vehicle (MI) or HA28 14 Days after Surgery HA28 at the dosage of 1 mg/kg was subcutaneously injected 1 day after LCA ligation once a day and continued for 14 days. *p < 0.05, **p < 0.01, ***p < 0.001 versus sham; ##p < 0.01, ###p < 0.001 versus vehicle. Molecular Therapy - Nucleic Acids 2019 14, 41-51DOI: (10.1016/j.omtn.2018.11.001) Copyright © 2018 The Authors Terms and Conditions

Figure 6 Effect of HA28 on Body Weight, Heart Rate, and Systolic Blood Pressure 14 Days after Surgery HA28 at the dosage of 1 mg/kg was subcutaneously injected 1 day after LCA ligation once a day and continued for 14 days. Molecular Therapy - Nucleic Acids 2019 14, 41-51DOI: (10.1016/j.omtn.2018.11.001) Copyright © 2018 The Authors Terms and Conditions

Figure 7 Changes in Echocardiographic Parameters in Sham-Operated Hamsters and MI Hamsters Treated with Vehicle (MI) or HA28 Numerals in parentheses indicate examined numbers of animals. EF, ejection fraction; FS, fractional shortening; Dd, left ventricular dimension end diastole; Ds, left ventricular dimension end systole; E:A, the ratio of the early (E) to late (A) ventricular filling velocities; E, E-wave velocity; A, A-wave velocity. *p < 0.05, **p < 0.01, ***p < 0.001 versus sham; #p < 0.05, ##p < 0.01, ###p < 0.001 versus vehicle. Molecular Therapy - Nucleic Acids 2019 14, 41-51DOI: (10.1016/j.omtn.2018.11.001) Copyright © 2018 The Authors Terms and Conditions