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Título

Rare non-coding APOB rs12713771 increases Lipid Droplet accumulation in HepG2 cells

Resumo

Background and Aims:
Non-coding variants refer to genetic variations that occur in regions of the genome that do not code for proteins. These variants can still have functional consequences by affecting gene expression, RNA splicing, or other regulatory processes. In APOB, these variants can disrupt the intracellular trafficking and processing of ApoB-containing lipoproteins, leading to dysregulated lipid metabolism and cholesterol accumulation. This study investigated the effects of the rare non-coding intronic variant APOB rs12713771 (c.4217-42G>A) on Lipid Droplet acumulation in HepG2 cells.

Methods:
LDL particle were isolated from carriers (A allele) and non-carriers (G allele) of APOB rs12713771 by sequential ultracentrifugation (1). LDL particles were measured by dynamic light scattering using Zetasizer NanoZS (Malvern Instruments, Malvern, UK). HepG2 cells (ATCC® HB-8065™) cultured in lipid deprivation medium for 24 hours and incubated with 15 ug/mL LDL for 0, 4 and 8 hours. Cells were fixed in a 4% paraformaldehyde solution overnight at 4˚ C, stained by Oil Red O (ORO) (2) and analyzed by light microscopy.

Results:
LDL - APOB rs12713771 (A allele) increase accumulation in lipid droplets compared to LDL control (G allele) in HepG2 cells in 0 and 4 hours of incubation (Table 1), and after 4 hours cells had more robust morphology and organization, indicating the internalization of LDL particles (Figure 1).

Conclusion:
The intronic APOBrs12713771 increases lipid accumulation in HepG2 cells, suggesting a deleterious effect on cholesterol homeostasis. Futher studies as needed to elucidate the impact of this variant on the intracellular trafficking and processing of ApoB-containing lipoprotein.

(1) Chapman, M. et al. Journal of Lipid Research 22, 339-358 (1981).
https://doi.org/10.1016/S0022-2275(20)35376-1.
(2) Mehlem, A., et al. Nat Protoc 8, 1149–1154 (2013).
https://doi.org/10.1038/nprot.2013.055.

Palavras Chave

Lipid droplets; Dislipidemia; APOB

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