ATP-Binding Cassette Transporter A1 Deficiency in Human Induced Pluripotent Stem Cell-Derived Hepatocytes Abrogates HDL Biogenesis and Enhances Triglyceride Secretion

Figures

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  Fig. 1. Impaired cholesterol efflux in TD HLCs. (A) Cholesterol efflux in the absence of exogenous apoA-I was compared between control-1 and TD-1 HLCs. All samples from the 3 clones of each group were compared. n = 9, * denotes p < 0.05. (B) Cholesterol efflux without (white bar) or with (black bar) addition of exogenous apoA-I (20 μg/ml) was compared for each group. n = 3, * denotes p < 0.05. Values are shown as mean ± SEM.

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  Fig. 2. Lack of nascent HDL formation in TD HLCs. (A and B) Gel-filtration chromatography profiles of nascent HDL formed in the absence of exogenous apoA-I based on 3H-choline-PL (A) and 14C-cholesterol (B) counts. (C and D) Gel-filtration chromatography profiles of nascent HDL formed in the presence of 20 μg/ml exogenous apoA-I based on 3H-choline-PL (C) and 14C-cholesterol (D) counts across fractions.

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  Fig. 3. Elevated TG secretion in TD vs. control HLCs. (A) Newly synthesized TG that was secreted into medium. Medium 3H-TG counts were normalized to media albumin mass. Data is presented as cpm 3H-TG/μg secreted albumin. (B) Percent of newly synthesized TG secretion. The percentage of TG secretion was calculated by dividing 3H-TG in the medium by the sum of counts in the medium and cells and multiplying by 100. All samples from the 3 clones of each group were compared. Values are shown as mean ± SEM. n = 8–9, * denotes p < 0.05.

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  Fig. 4. Transcriptome profiles of HLCs and validation of ANGPTL3. (A) The overlap of genes expressed in TD-1 and control-1 HLCs was evaluated by Pearson Correlation Coefficient. (B) Heat map of pluripotency and hepatic markers between groups. (C) Heat map of differentially expressed lipid-related genes according to Gene Ontology lipid terms. (D) Expression of ANGPTL3 measured by RT-PCR. (E) Culture medium ANGPTL3 protein concentrations determined by ELISA. (F) Plasma ANGPTL3 levels were measured by ELISA. Values are shown as mean ± SEM. n = 9 for RT-PCR analysis. n = 3 for culture medium ELISA. n = 4–19 for plasma ANGPTL3 ELISA. * denotes p < 0.05.