June 2013
Volume 54, Issue 15
Free
ARVO Annual Meeting Abstract  |   June 2013
COMPARISON OF THE GENE EXPRESSION BY HAPLOGROUPS H AND J: IMPLICATIONS FOR AMD (AGE RELATED MACULAR DEGENERATION)
Author Affiliations & Notes
  • Claudio Ramirez
    Gavin Herbert Eye Institute, University of California, Irvine, Irvine, CA
  • Marilyn Chwa
    Gavin Herbert Eye Institute, University of California, Irvine, Irvine, CA
  • Shari Atilano
    Gavin Herbert Eye Institute, University of California, Irvine, Irvine, CA
  • Deepika Malik
    Gavin Herbert Eye Institute, University of California, Irvine, Irvine, CA
  • Javier Cáceres del Carpio
    Gavin Herbert Eye Institute, University of California, Irvine, Irvine, CA
  • Mohamed Tarek
    Gavin Herbert Eye Institute, University of California, Irvine, Irvine, CA
  • S Michal Jazwinski
    Tulane Center for Aging, Tulane University, New Orleans, LA
  • Miceli Michael
    Tulane Center for Aging, Tulane University, New Orleans, LA
  • Baruch Kuppermann
    Gavin Herbert Eye Institute, University of California, Irvine, Irvine, CA
  • Cristina Kenney
    Gavin Herbert Eye Institute, University of California, Irvine, Irvine, CA
  • Footnotes
    Commercial Relationships Claudio Ramirez, None; Marilyn Chwa, None; Shari Atilano, None; Deepika Malik, None; Javier Cáceres del Carpio, None; Mohamed Tarek, None; S Michal Jazwinski, None; Miceli Michael, None; Baruch Kuppermann, Alimera (C), Allegro (C), Allergan (C), Genentech (C), Glaukos (C), GSK (F), Novagali (C), Novartis (C), Ophthotech (C), Pfizer (C), Regeneron (C), Santen (C), SecondSight (C), Teva (C), ThromboGenics (C); Cristina Kenney, None
  • Footnotes
    Support None
Investigative Ophthalmology & Visual Science June 2013, Vol.54, 4985. doi:https://doi.org/
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      Claudio Ramirez, Marilyn Chwa, Shari Atilano, Deepika Malik, Javier Cáceres del Carpio, Mohamed Tarek, S Michal Jazwinski, Miceli Michael, Baruch Kuppermann, Cristina Kenney; COMPARISON OF THE GENE EXPRESSION BY HAPLOGROUPS H AND J: IMPLICATIONS FOR AMD (AGE RELATED MACULAR DEGENERATION). Invest. Ophthalmol. Vis. Sci. 2013;54(15):4985. doi: https://doi.org/.

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

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Abstract

Purpose: AMD is a leading cause of vision loss in the elderly population. Mitochondrial genetics is an important area of research that may help us understand the predisposition for AMD between and within racial groups.Previous studies have shown that the mtDNA haplogroup J is associated with AMD, while the H haplogroup is protective. However, the functional consequences of this difference are not understood. We have used cybrids (cytoplasmic hybrids) to study the characteristics and biochemical differences between various haplogroups. Our hypothesis is that cybrids, dissimilar only in their mtDNA haplogroup will behave differently in vitro.

Methods: Cybrids were created by introducing the mitochondria from human individuals platelets into a host cell line (ARPE-19) that was devoid of mitochondrial DNA (Rho0). Therefore, all cybrids carry the same nuclear genes but vary only in their mitochondrial content. Cybrid cultures H and J were pelleted, the RNA was isolated and then quantified. RNA samples were reverse transcribed into cDNA. The RNAs from the three H and the three J haplogroups cybrid cultures were combined into a single sample each for analyses with the Affymetrix Human U133 Plus 2.0 Array. The gene expression results were analyzed with pathway analysis software (INGENUITY Systems). Q-PCR was performed using primers for genes associated with inflammation (TGFA.TGFB2, and IL6) and apoptosis (RARA1, BBC-3 and BCL2L13).

Results: The array analyses showed that H and J cybrids had altered expression of nuclear genes involved in inflammation and apoptotic pathways. Q-PCR analyses showed that J cybrids had decreased expression levels for TGFA (0.43 fold, p=0.03), RARA (0.57 fold, p=0.007), and BLC2L13 (0.56 fold, p=0.005). There were no significant changes in expression levels for TGFB2 (0.86 fold, p=0.41), IL-6 (0.38 fold, p=0.09), and BBC-3 (0.70 fold, p=0.11) in the H versus J cybrids.

Conclusions: This study demonstrates that cybrids may be a useful tool to study the effects of mtDNA variants on gene expression. Our findings suggest that mtDNA haplogroup differences have functional consequences. This data may have implications for understanding the association between haplogroup J and its increased risk for AMD.

Keywords: 412 age-related macular degeneration • 533 gene/expression • 600 mitochondria  
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