September 2016
Volume 57, Issue 12
Open Access
ARVO Annual Meeting Abstract  |   September 2016
Comparison of Lipid and Cholesterol-Biosynthesis Gene Products in H- and K-mt-DNA haplogroup cybrids
Author Affiliations & Notes
  • Christine Garabetian
    Gavin Herbert Eye Institute, University of California Irvine, Irvine, California, United States
  • Kunal Thaker
    Gavin Herbert Eye Institute, University of California Irvine, Irvine, California, United States
  • Javier Cáceres-del-Carpio
    Gavin Herbert Eye Institute, University of California Irvine, Irvine, California, United States
  • Shari R Atilano
    Gavin Herbert Eye Institute, University of California Irvine, Irvine, California, United States
  • Marilyn Chwa
    Gavin Herbert Eye Institute, University of California Irvine, Irvine, California, United States
  • Anthony B Nesburn
    Gavin Herbert Eye Institute, University of California Irvine, Irvine, California, United States
    Cedars-Sinai Medical Center, Los Angeles, California, United States
  • Cristina M Kenney
    Gavin Herbert Eye Institute, University of California Irvine, Irvine, California, United States
    Department of Pathology and Laboratory Medicine, University of California Irvine, Irvine, California, United States
  • Footnotes
    Commercial Relationships   Christine Garabetian, None; Kunal Thaker, None; Javier Cáceres-del-Carpio, None; Shari Atilano, None; Marilyn Chwa, None; Anthony Nesburn, None; Cristina Kenney, None
  • Footnotes
    Support  Discovery Eye Foundation, Guenther Foundation, Beckman Initiative for Macular Research, Polly and Michael Smith Foundation, Max Factor Family Foundation, Iris and B. Gerald Cantor Foundation, J.C.d.C. was the David & Julianna Pyott Pan-American-Retina Research Fellowship
Investigative Ophthalmology & Visual Science September 2016, Vol.57, 6054. doi:
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    • Get Citation

      Christine Garabetian, Kunal Thaker, Javier Cáceres-del-Carpio, Shari R Atilano, Marilyn Chwa, Anthony B Nesburn, Cristina M Kenney; Comparison of Lipid and Cholesterol-Biosynthesis Gene Products in H- and K-mt-DNA haplogroup cybrids. Invest. Ophthalmol. Vis. Sci. 2016;57(12):6054.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract

Purpose : Cholesterol has been associated in the pathogenesis of neurodegenerative and systemic pathologies, and certain populations have higher incidence of cholesterol-linked familial diseases. The purpose of this experiment was to determine if mitochondrial DNA (mt-DNA) haplogroup H and K transmitochondrial cybrids regulate the cholesterol and lipid biosynthesis pathways differently. Using transmitochondrial cybrids, cell lines with identical nuclei but either H or K mtDNA, we looked at the relationship between the gene expression levels of cholesterol-related genes and neutral lipid deposits in H and K cybrids.

Methods : Transmitochondrial cybrids were created by fusing mitochondria (mt) isolated from human platelets and cells lacking mtDNA; thus these cybrids have identical nuclei but either H or K mtDNA. RNA from H and K cybrids were analyzed using GeneChip arrays (n=3). Specific cholesterol-related genes FDFT1, SQLE, LSS, CYP51A1, EBP, and DHCR24 were analyzed using Q-PCR (H, n=7; K, n=9). Corresponding reference genes were used as housekeepers for each of the six genes studied. H and K cybrids (n=10) were seeded at 104/well and stained with HCS LipidTox Green Neutral Lipid Stain. Statistical analyses were performed using student’s t-test.

Results : Q-PCR showed insignificant differences of the cholesterol biosynthesis genes in H vs. K; however, one K cybrid (CY#13-57) showed elevated expression of 4 out of 6 cholesterol biosynthesis genes compared to H cybrids; (17.4-fold increase in FDFT1 expression, a 31-fold increase in SQLE expression, a 10.8-fold increase in CYP51A1 expression, and a 5.4-fold increase in EBP). LipidTox staining showed cytoplasmic distribution of the lipids with significantly elevated intensity in the CY#13-57 compared to H cybrids or other K cybrids (p<0.001).

Conclusions : Due to all cybrids differing only in their mtDNA content, our findings suggest that H- and K-haplogroup mt-DNA do not necessarily affects genes involved in cholesterol biosynthesis. However, the increased expression of cholesterol-related genes and larger and more vibrant lipid deposits seen in CY#13-57 suggest the mt-DNA in that specific K cybrid is different than other K cybrids and is mediating cholesterol production and lipid transport through some mechanism.

This is an abstract that was submitted for the 2016 ARVO Annual Meeting, held in Seattle, Wash., May 1-5, 2016.

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