Aging
Navigate
Submit an Article
Research Paper|Volume 8, Issue 5|pp 945—957

Study of mitochondrial respiratory defects on reprogramming to human induced pluripotent stem cells

Sandy S.C Hung1, Nicole J Van Bergen1, Stacey Jackson1, Helena Liang1, David A. A Mackey2, Damián Hernández3, Shiang Y Lim1,3, Alex W Hewitt1,4, Ian Trounce1, Alice Pébay1, Raymond C.B Wong1
  • 1Centre for Eye Research Australia, Royal Victorian Eye and Ear Hospital and Ophthalmology, Department of Surgery, The University of Melbourne, Melbourne, Australia
  • 2Lions Eye Institute and University of Western Australia, Nedlands, Australia
  • 3O'Brien Institute Department, St Vincent's Institute of Medical Research, Fitzroy, Australia
  • 4School of Medicine, Menzies Institute for Medical Research, University of Tasmania, Tasmania, Australia
Received: March 3, 2016Accepted: April 17, 2016Published: April 26, 2016
https://doi.org/10.18632/aging.100950

Copyright: © 2016 Hung et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

Abstract

Reprogramming of somatic cells into a pluripotent state is known to be accompanied by extensive restructuring of mitochondria and switch in metabolic requirements. Here we utilized Leber's hereditary optic neuropathy (LHON) as a mitochondrial disease model to study the effects of homoplasmic mtDNA mutations and subsequent oxidative phosphorylation (OXPHOS) defects in reprogramming. We obtained fibroblasts from a total of 6 LHON patients and control subjects, and showed a significant defect in complex I respiration in LHON fibroblasts by high-resolution respiratory analysis. Using episomal vector reprogramming, our results indicated that human induced pluripotent stem cell (hiPSC) generation is feasible in LHON fibroblasts. In particular, LHON-specific OXPHOS defects in fibroblasts only caused a mild reduction and did not significantly affect reprogramming efficiency, suggesting that hiPSC reprogramming can tolerate a certain degree of OXPHOS defects. Our results highlighted the induction of genes involved in mitochondrial biogenesis (TFAM, NRF1), mitochondrial fusion (MFN1, MFN2) and glycine production (GCAT) during reprogramming. However, LHON-associated OXPHOS defects did not alter the kinetics or expression levels of these genes during reprogramming. Together, our study provides new insights into the effects of mtDNA mutation and OXPHOS defects in reprogramming and genes associated with various aspects of mitochondrial biology.