220 Chapter 8 Figure 6: Genetic engineering and agonist therapy as novel treatment strategies targeting dysregulated lipid metabolism in PLN-R14del cardiomyopathy. A) Immunofluorescence images of Nile red staining showing the lipid accumulation in all hiPSC-CMs groups. Nuclei were stained by DAPI (blue). Bar graph showing a significantly higher intracellular lipid formation in PLN-R14del hiPSC-CMs when compared to the healthy controls as well as the isogenic controls in which PLN-R14del was corrected (n ≥ 4 per group), whereas the lipid accumulation in hiPSC-CMs from PLN-R14del patient, an asymptomatic PLN-R14del carrier, and a homozygous PLN-R14del group reamined comparable. Data were normalised to nuclei and shown as mean ±SEM. One-way ANOVA was used, **P<0.01, ***P<0.001. B) Ca2+ transient parameters for examining the Ca2+ 590 handling property in hiPSC-CMs, including rise time (RT), decay time (DT), calcium full-width half maximum (FWHM), duration at 25% decline from maximum amplitude (CTD25), and duration at 75% decline from maximum amplitude (CTD75). Representative calcium transient in control and PLN-R14del hiPSC-CMs with and without bezafibrate treatment (dashed and solid line, respectively). Each trace is an average DF/F 0 versus time plot (33 Hz) in each well and three wells per condition per group were included in this analysis. C) Immunofluorescence staining of HADHA (red) and HADHB (green) in control and PLN-R14del hiPSC-CMs with and without bezafibrate treatment. Nuclei were stained by DAPI (blue). All images were taken at 63x magnification. Bar graphs showing significantly elevated HADHA and HADHB levels after bezafibrate treatment in PLN-R14del, but not in the control hiPSC-CMs. Data was normalised to the number of nuclei. Each dot represents the normalized data per obtained images and shown as mean ±SD. Two-way ANOVA was used in the bezafibrate treatment experiment, ns= not significant, **P<0.01, ***P<0.001, ****P<0.0001. D) Schematic overview of PPARA-mediated FAO dysregulation and impaired lipid metabolism in PLN-R14del versus control cardiomyocytes
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