June 2013
Volume 54, Issue 15
Free
ARVO Annual Meeting Abstract  |   June 2013
Comparison Between Haplogroup H and L Cybrids in ROS/RNS Production and Gene Expression Further step in understanding the pathogenesis of Age-related Macular Degeneration
Author Affiliations & Notes
  • Mohamed Tarek
    Gavin Herbert Eye Institute, Irvine, CA
  • Javier Cáceres del Carpio
    Gavin Herbert Eye Institute, Irvine, CA
  • Claudio Ramirez
    Gavin Herbert Eye Institute, Irvine, CA
  • Payam Falatoonzadeh
    Gavin Herbert Eye Institute, Irvine, CA
  • Shari Atilano
    Gavin Herbert Eye Institute, Irvine, CA
  • Deepika Malik
    Gavin Herbert Eye Institute, 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, Irvine, CA
  • Cristina Kenney
    Gavin Herbert Eye Institute, Irvine, CA
  • Footnotes
    Commercial Relationships Mohamed Tarek, None; Javier Cáceres del Carpio, None; Claudio Ramirez, None; Payam Falatoonzadeh, None; Shari Atilano, None; Deepika Malik, 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, 6195. doi:https://doi.org/
  • Views
  • Share
  • Tools
    • Alerts
      ×
      This feature is available to authenticated users only.
      Sign In or Create an Account ×
    • Get Citation

      Mohamed Tarek, Javier Cáceres del Carpio, Claudio Ramirez, Payam Falatoonzadeh, Shari Atilano, Deepika Malik, S Michal Jazwinski, Miceli Michael, Baruch Kuppermann, Cristina Kenney; Comparison Between Haplogroup H and L Cybrids in ROS/RNS Production and Gene Expression Further step in understanding the pathogenesis of Age-related Macular Degeneration. Invest. Ophthalmol. Vis. Sci. 2013;54(15):6195. doi: https://doi.org/.

      Download citation file:


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

      ×
  • Supplements
Abstract

Purpose: Mitochondrial DNA haplogroups can be classified according to accumulation of specific single nucleotide polymorphisms (SNPs). The H haplogroup is of European origin and protective for AMD, the L haplogroup which is of African origin shows only early AMD changes. Our work is to investigate the differences in Reactive Oxygen/Nitrogen Species (ROS/RNS) production and gene expression profiles of cybrids (cytoplasmic hybrids) that have either the H haplogroup or the L haplogroup.

Methods: ARPE-19 cells lacking mitochondrial DNA (mtDNA) (Rho0) were fused with platelets from individuals with either H or L haplogroup mtDNA as determined by PCR analysis. After culture for 24 hours, ROS/RNS levels were determined with 2`, 7`-dichlorodihydrofluorescein diacetate dye assay. Experiments were repeated twice and performed in 8 wells per sample. RNA was obtained from the H cybrids (n=3) and L cybrids (n=3) and the gene expression patterns were determined using the Affymetrix Human Genome U133 plus 2.0 array. The pathway profiles were analyzed with the INGENUITY systems program. Quantitative PCR (Q-PCR) was performed using primers for complement component 3 (C3) and complement factor H (CFH) genes for the different H and L cybrids.

Results: Cybrids with the haplogroup L mtDNA produced 25.79±6.21 % lower levels of ROS/RNS compared to haplogroup H cybrids (p=0.026). Complement pathway genes were expressed differently, the haplogroup H cybrids showed higher expression levels in C1S, C3, CFH, C4BPB, CFHR4, and ITGB2 genes by 2.9, 5.5, 2, 3.8, 17.8 and 3.4 folds respectively, while the C1QC, C4A, C8B and CFP genes were expressed at higher levels in the haplogroup L cybrids (3.2, 3.3, 6.2 and 3 folds respectively). By Q-PCR analyses, the L cybrids had a 0.66-fold expression level for the CFH gene (p=0.03) and 0.17-fold level for the C3 gene (p=0.0004) compared to the H cybrids.

Conclusions: Our findings demonstrate that cybrids which are identical except for their mtDNA, have different levels of expression for genes involved in the alternative complement pathway. Our results are significant because they show that mtDNA variants (haplogroups) can mediate not only energy production but also the expression of genes for major cellular pathways involved in inflammation.

Keywords: 412 age-related macular degeneration • 539 genetics • 600 mitochondria  
×
×

This PDF is available to Subscribers Only

Sign in or purchase a subscription to access this content. ×

You must be signed into an individual account to use this feature.

×