April 2011
Volume 52, Issue 14
Free
ARVO Annual Meeting Abstract  |   April 2011
Rescue Of Cybrids Containing Leber Hereditary Optic Neuropathy (LHON) Mutation Using Bax-inhibiting Peptide
Author Affiliations & Notes
  • Dan-Ning Hu
    Pathology & Ophthalmology,
    New York Eye & Ear Infirmary, New York, New York
  • Ian Chan
    Ophthalmology,
    New York Eye & Ear Infirmary, New York, New York
  • Min Chen
    Pathology,
    New York Eye & Ear Infirmary, New York, New York
  • Steven A. McCormick
    Pathology,
    New York Eye & Ear Infirmary, New York, New York
  • Joseph Walsh
    Ophthalmology,
    New York Eye & Ear Infirmary, New York, New York
  • Footnotes
    Commercial Relationships  Dan-Ning Hu, None; Ian Chan, None; Min Chen, None; Steven A. McCormick, None; Joseph Walsh, None
  • Footnotes
    Support  New York Eye & Ear Infirmary, Ophthalmology Research Funds
Investigative Ophthalmology & Visual Science April 2011, Vol.52, 5440. doi:
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    • Get Citation

      Dan-Ning Hu, Ian Chan, Min Chen, Steven A. McCormick, Joseph Walsh; Rescue Of Cybrids Containing Leber Hereditary Optic Neuropathy (LHON) Mutation Using Bax-inhibiting Peptide. Invest. Ophthalmol. Vis. Sci. 2011;52(14):5440.

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

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Abstract
 
Purpose:
 

To investigate the use of Bax-inhibiting peptide V5 (BIP) to block apoptosis in cybrids containing the LHON mutation. BIP is derived from the amino acid sequence of the Bax-binding domain of Ku70 and blocks the Bax-Bak oligomerization on outer mitochondria membranes.

 
Methods:
 

Homeoplasmic 143B osteosarcoma cybrids containing the mtDNA (G11778A) mutation were grown in Dulbecco’s Modified Eagle Medium (DMEM) with high glucose supplemented with 10% fetal bovine serum. Cells were seeded into 96 well plates and cultured with DMEM, glucose-free galactose medium (GFGM) and GFGM with BIP at 10, 30, 100 and 300 µmol concentrations. After 48 hrs, 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT, 50 µl of 1 mg/ml) was added, incubated for 4 hrs, and replaced by DMSO. A microplate spectrophotometer at 540 nm was used to evaluate cell survival.

 
Results:
 

Compared to cybrids in DMEM medium, cell viability from MTT assay showed that cybrid survival % dropped to 34% after incubation in GFGM after 48 hours. Adding BIP to GFGM at 10, 30, 100 and 300 µmol concentrations increased survival in a dose dependent fashion to 42%, 48%, 50% and 56%, respectively, after 48 hrs. P-values for 2-tailed t-test of cell survival for BIP at concentrations of 10, 30, 100 and 300 µmol with GFGM vs. GFGM alone were 0.02, 0.01, 0.0002 and 0.0001 respectively.

 
Conclusions:
 

BIP demonstrated statistically significant dose-dependent protective effects in the LHON cybrid model.  

 
Keywords: neuro-ophthalmology: optic nerve 
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