Journal: Nature Cardiovascular Research

Article Title: Plakophilin 2 gene therapy prevents and rescues arrhythmogenic right ventricular cardiomyopathy in a mouse model harboring patient genetics

doi: 10.1038/s44161-023-00370-3

Figure Lengend Snippet: ( a ) Representative telemetry ECG tracings and premature ventricular contraction (PVC) counts from Ctrl and PKP2 heterozygous (Het) mice at 3 months of age. Red arrows indicate premature ventricular contractions. Scale bar = 0.1 s. Data are presented as mean ± S.E.M. n = 3 biologically independent animals examined per group, two-tailed unpaired t test, * p < 0.05 (p = 0.0135). ( b ) Quantification of end-diastolic volumes (EDV), end-systolic volumes (ESV) and percentage of ejection fraction (EF %) in WT and PKP2 Het mice at 10 months old of age using cardiac MRI. n = 4 biologically independent animals examined per group. Data are presented as mean ± S.E.M. Two-way ANOVA with Tukey’s multiple comparison test.

Article Snippet: For MRI image analyses, two-dimensional endocardial contours were manually segmented and slice volumes and/or ejection fractions calculated using freely available software (Segment CMR version 4.0 R12067, MEDVISO) for each heart at end-diastole and end-systole (both left and right ventricle).

Techniques: Two Tailed Test, Comparison

Journal: Nature Cardiovascular Research

Article Title: Plakophilin 2 gene therapy prevents and rescues arrhythmogenic right ventricular cardiomyopathy in a mouse model harboring patient genetics

doi: 10.1038/s44161-023-00370-3

Figure Lengend Snippet: a , Schemata for early injection of AAV-GFP or AAV-PKP2 to PKP2 Hom mice at postnatal day 2 and post analysis at 6 months. b , Kaplan–Meier survival analysis (log-rank test) of mice (controls, n = 18; Hom, n = 23; Hom-AAV-PKP2, n = 7 biologically independent animals). Ctrl and PKP2 Hom data in Fig. 7 are also presented in Fig. . **** P < 0.0001 ( Χ 2 58.07, d.f. 2). c , Western blot analysis of PKP2 and cell–cell junctional proteins (DSP, DSG2, JUP, N-Cad and CX43) in mouse hearts. GAPDH was used as a loading control. d , Quantification of protein expression shown in c normalized to GAPDH. Data are presented as mean ± s.e.m., n = 5 biologically independent animals. Two-way ANOVA with Sidak’s multiple comparison test. ****Adjusted P < 0.0001. e , Representative short-axis cardiac MRI views. f , Quantification of heart rate and ejection fraction in mice using cardiac MRI (controls, n = 4; Hom-AAV-GFP, n = 6; Hom-AAV-PKP2, n = 5 biologically independent animals). Data are presented as mean ± s.e.m. One-way ANOVA for heart rate comparison. Two-way ANOVA with Bonferroni’s multiple comparison test for other comparisons. Adjusted P values, **** P < 0.0001, * P < 0.05 ( P = 0.0358). Historical data for Hom-AAV-GFP are from 6 weeks as no PKP2 Hom mouse survived to 6 months of age. g , Representative composite surface ECG tracings averaged from four beats in untreated wild-type control and PKP2 Hom mice treated with AAV-PKP2. Scale bar, 10 ms. h , Quantification of heart rate and QRS intervals from composite surface ECG tracings (controls, n = 5; Hom-AAV-PKP2, n = 6 biologically independent animals). Data are presented as mean ± s.e.m. Two-tailed unpaired t -test. NS, not significant. i , Representative ECG tracings from untreated control and PKP2 Hom mice treated with AAV-PKP2. j , Blood serum analysis for ALT and ALP liver enzyme levels in untreated wild-type control and PKP2 Hom mice treated with AAV-PKP2 (controls, n = 5; Hom-AAV-PKP2, n = 6 biologically independent animals). Normal enzyme limits are indicated with dotted lines. Data are presented as mean ± s.e.m. Two-tailed unpaired t -test.

Article Snippet: For MRI image analyses, two-dimensional endocardial contours were manually segmented and slice volumes and/or ejection fractions calculated using freely available software (Segment CMR version 4.0 R12067, MEDVISO) for each heart at end-diastole and end-systole (both left and right ventricle).

Techniques: Injection, Western Blot, Control, Expressing, Comparison, Two Tailed Test

Journal: Nature Cardiovascular Research

Article Title: Plakophilin 2 gene therapy prevents and rescues arrhythmogenic right ventricular cardiomyopathy in a mouse model harboring patient genetics

doi: 10.1038/s44161-023-00370-3

Figure Lengend Snippet: a , Schemata for late injection of AAV-GFP or AAV-PKP2 to PKP2 Hom mice at 4 weeks and post analysis at 6 weeks. gc, genom copies. b , Western blot analysis of PKP2 and desmosomal cell–cell junctional proteins (DSP, DSG2 and JUP) in mouse hearts. GAPDH was used as a loading control. Experiments were repeated independently three times with similar results. c , Representative short-axis MRI views from mice. d , Quantification of heart rate and ejection fraction in mice using cardiac MRI ( n = 6 biologically independent animals). Data are presented as mean ± s.e.m. Two-way ANOVA with Sidak’s multiple comparison test. ** P < 0.01 ( P = 0.0026), * P < 0.05 ( P = 0.0450). e , Schemata for late injection of AAV-GFP or AAV-PKP2 to PKP2 Hom mice at 4 weeks and post analysis at 20 weeks. gc, genom copies. f , Kaplan–Meier survival analysis (log-rank test) of mice (controls, n = 8; Hom-AAV-GFP, n = 8; Hom-AAV-PKP2, n = 7 biologically independent animals. ***adjusted P < 0.001 ( Χ 2 16.59, d.f. 2, P = 0.0003).

Article Snippet: For MRI image analyses, two-dimensional endocardial contours were manually segmented and slice volumes and/or ejection fractions calculated using freely available software (Segment CMR version 4.0 R12067, MEDVISO) for each heart at end-diastole and end-systole (both left and right ventricle).

Techniques: Injection, Western Blot, Control, Comparison

Journal: Nature Cardiovascular Research

Article Title: Plakophilin 2 gene therapy prevents and rescues arrhythmogenic right ventricular cardiomyopathy in a mouse model harboring patient genetics

doi: 10.1038/s44161-023-00370-3

Figure Lengend Snippet: ( a ) Schemata for late injection of AAV GFP or AAV PKP2 to PKP2 Hom mice at 4 weeks and post-analysis at 9 weeks (top). Western blot analysis of PKP2 and desmosomal proteins (DSP, DSG2 and JUP) in mouse hearts. β-actin used as a loading control. Data are presented as mean ± S.E.M. n = 6 biologically independent animals per group, Two-way ANOVA with Tukey’s multiple comparison test. *, p < 0.05 (PKP2, p = 0.0410), **, p < 0.01 (DSG2, p = 0.0099; JUP, p = 0.0020). (b) Representative Masson’s Trichrome stains from left (LV) and right ventricular (RV) sections from 9-week-old Ctrl, PKP2 Hom-AAV GFP (five weeks post-late stage administration) and PKP2 Hom-AAV PKP2 (five weeks post-late stage administration) mice. Scale bar, 2 mm. Experiments were repeated independently three times with similar results. ( c ) Quantification of mice demonstrating PVCs and sudden death at 9 weeks of age in Ctrl, PKP2 Hom-AAV GFP (five weeks post-late stage administration) and PKP2 Hom-AAV PKP2 (five weeks post-late stage administration) mice. (d) Quantification of end-diastolic volumes (EDV), end-systolic volumes (ESV) and percentage of ejection fraction (EF %) in 9 week old Ctrl, PKP2 Hom-AAV GFP (five weeks post-late stage administration) and Hom-AAV PKP2 (five weeks post-late stage administration) mice using cardiac MRI. Data are presented as mean ± S.E.M, n = 4 for Ctrl, n = 5 for Hom-AAV GFP group and n = 6 for Hom-AAV PKP2 biologically independent animals. Two-way ANOVA with Tukey’s multiple comparison test. ****, adjusted p < 0.0001. ***, p < 0.001 (EF, LV, p = 0.0008; RV, p = 0.0008). *, p < 0.05 (EDV, LV, p = 0.0369; RV, p = 0.0148; ESV, LV, p = 0.0352), ns, not significant.

Article Snippet: For MRI image analyses, two-dimensional endocardial contours were manually segmented and slice volumes and/or ejection fractions calculated using freely available software (Segment CMR version 4.0 R12067, MEDVISO) for each heart at end-diastole and end-systole (both left and right ventricle).

Techniques: Injection, Western Blot, Control, Comparison

Journal: Journal of the American Heart Association: Cardiovascular and Cerebrovascular Disease

Article Title: Contrast‐Enhanced Cardiac Magnetic Resonance Imaging With a Manganese‐Based Alternative to Gadolinium for Tissue Characterization of Acute Myocardial Infarction

doi: 10.1161/JAHA.122.026923

Figure Lengend Snippet: Model validation and study protocol overview. We performed 40 to 80 minutes of left anterior descending coronary artery (LAD) occlusion just distal to the second diagonal branch. Delayed enhancement imaging was performed with both contrast agents. In 1 animal subject, triphenyltetrazolium chloride (TTC) staining was performed for model validation. The gross pathological specimen and best‐matching cardiac magnetic resonance imaging (CMR) planes are presented in the top row. T2 was prolonged in the lesioned area, suggestive of edema (bottom right). The green inlay details our imaging protocol. Representative late gadolinium enhancement CMR images and the following segmentation are provided (red=endocardial border, green=epicardial border, and yellow=lesion border). Gd‐DOTA indicates gadoteric acid; and Myo, myocardium.

Article Snippet: Image analysis was performed using the software Medviso Segment CMR version 10187b ( http://segment.heiberg.se ).

Techniques: Biomarker Discovery, Imaging, Staining, Magnetic Resonance Imaging