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Energy and Macronutrient Metabolism (Poster Session)

(P04-002-25) Differentiated ARPE-19 Cells Show Variation in Gene Expression Involved in Lipid Utilization When Transformed With mFAT-1

Saturday, May 31, 2025
11:45 AM - 12:45 PM ET
Location: Poster Board B-2

    Presenting Author(s)

  • HB

    Henry H. Brak

    Student
    Lake Erie College of Osteopathic Medicine
    Erie, Pennsylvania, United States

    • Co-Author(s)

  • PB

    Peter M. Baldwin

    Lake Erie College of Osteopathic Medicine, United States

  • BJ

    Bo Song Jang

    Lake Erie College of Osteopathic Medicine, United States

  • RG

    Ryan C. Garich

    Lake Erie College of Osteopathic Medicine, United States

  • DD

    David M. Duriancik, PhD

    Associate Professor of Biochemistry
    Lake Erie College of Osteopathic Medicine

Disclosure(s):
Henry H. Brak: No relevant financial relationship(s) with ineligible companies to disclose.


Objectives: Previous investigations have shown that ARPE-19 cells significantly alter gene expression after differentiation which is particularly important when using ARPE-19 cells as an accurate in vitro model of human pigmented retinal epithelium. Furthermore, ARPE-19 cells are unable to endogenously produce n-3 polyunsaturated fatty acids (PUFA) at levels that are available in the human body. This study investigates the effects differentiation and n-3 PUFA availability have on genes important in lipid and fat-soluble vitamin utilization in ARPE-19 cells.

Methods: ARPE-19 cells were transformed with C. elegans mFAT-1 gene to induce n-3 PUFA synthesis. Cells were cultured for 4 months to induce differentiation. RNA was isolated using Trizol reagent and cDNA was synthesized for qPCR. Relative expression of genes important for lipid and fat-soluble vitamin utilization was determined and compared against cultures that have not differentiated and/or were not transformed with the mFAT-1 transgene.

Results: Differentiated ARPE-19 cells showed significantly increased expression of DCT and PLTP while also displaying decreased expression of TTPA, FASN and FADS2 compared to undifferentiated ARPE-19 cells. mFAT-1 caused decreased expression of STRA6, TTPA, PLTP, FASN, and FADS2 in undifferentiated ARPE-19 cells. This effect decreased at 4 months and only DCT, PLTP and FADS2 exhibit a significant decrease in expression when compared to cells without mFAT-1.

Conclusions: Differentiation of ARPE-19 cells significantly alters fat-soluble vitamin utilization and fatty acid metabolism. Additionally, increased availability of n-3 PUFA decreases expression of endogenous genes important in phospholipid formation and fatty acid desaturation. These findings highlight important considerations researchers can implement to more accurately model the human retina using in vitro cell culture.

Funding Sources: Internal

Funding: Lake Erie College of Osteopathic Medicine