Abstract
Leber’s Hereditary Optic Neuropathy is the most prevalent mitochondrial neurological disease caused by mutations in mitochondrial DNA encoded respiratory complex I subunits. Although the genetic origin for Leber’s hereditary optic neuropathy was identified about 30 years ago, the underlying pathogenesis is still unclear primarily due to the lack of a relevant system or cell model. Current models are limited to lymphoblasts, fibroblasts, or cybrid cell lines. As the disease phenotype is limited to retinal ganglion cells, induced pluripotent stem cells will serve as an excellent model for studying this tissue-specific disease, elucidating its underlying molecular mechanisms, and identifying novel therapeutic targets. Here, we describe a detailed protocol for the generation of retinal ganglion cells, and also cardiomyocytes for proof of iPSC pluripotency.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Similar content being viewed by others
References
Al-Enezi M, Al-Saleh H, Nasser M (2008) Mitochondrial disorders with significant ophthalmic manifestations. Middle East Afr J Ophthalmol 15:81–86. https://doi.org/10.4103/0974-9233.51998
Han J, Lee Y-M, Kim SM, Han SY, Lee JB, Han S-H (2015) Ophthalmological manifestations in patients with Leigh syndrome. Br J Ophthalmol 99:528–535. https://doi.org/10.1136/bjophthalmol-2014-305704
Wallace DC, Lott MT (2017) Leber hereditary optic neuropathy: exemplar of an mtDNA disease. Handb Exp Pharmacol 240:339–376. https://doi.org/10.1007/164_2017_2
Wallace D, Singh G, Lott M, Hodge J, Schurr T, Lezza A, Elsas L, Nikoskelainen E (1988) Mitochondrial DNA mutation associated with Leber’s hereditary optic neuropathy. Science 242:1427–1430. https://doi.org/10.1126/science.3201231
Yu-Wai-Man P, Turnbull DM, Chinnery PF (2002) Leber hereditary optic neuropathy. J Med Genet 39:162–169. https://doi.org/10.1136/jmg.39.3.162
Bahr T, Welburn K, Donnelly J, Bai Y (1866) Emerging model systems and treatment approaches for Leber’s hereditary optic neuropathy: challenges and opportunities. Biochim Biophys Acta 2020:165743. https://doi.org/10.1016/j.bbadis.2020.165743
Danielson SR, Wong A, Carelli V, Martinuzzi A, Schapira AHV, Cortopassi GA (2002) Cells bearing mutations causing Leber’s hereditary optic neuropathy are sensitized to Fas-induced apoptosis. J Biol Chem 277:5810–5815. https://doi.org/10.1074/jbc.M110119200
Ghelli A, Zanna C, Porcelli AM, Schapira AHV, Martinuzzi A, Carelli V, Rugolo M (2003) Leber’s hereditary optic neuropathy (LHON) pathogenic mutations induce mitochondrial-dependent apoptotic death in transmitochondrial cells incubated with galactose medium. J Biol Chem 278:4145–4150. https://doi.org/10.1074/jbc.M210285200
Martikainen M (2012) Mitochondrial disease in southwestern Finland. Population-based molecular genetic and clinical studies. In: Undefined. /paper/mitochondrial-disease-in-southwestern-Finland.-and-Martikainen/d8bfb057352eba26ee609e718475bbd3c0fb8055. Accessed 9 Dec 2020
DiMauro S, Davidzon G (2005) Mitochondrial DNA and disease. Ann Med 37:222–232. https://doi.org/10.1080/07853890510007368
Li Y, Nguyen HV, Tsang SH (2016) Skin biopsy and patient-specific stem cell lines. Methods Mol Biol 1353:77–88. https://doi.org/10.1007/7651_2015_225
Vangipuram M, Ting D, Kim S, Diaz R, Schüle B (2013) Skin punch biopsy explant culture for derivation of primary human fibroblasts. J Vis Exp. https://doi.org/10.3791/3779
Kime C, Rand TA, Ivey KN, Srivastava D, Yamanaka S, Tomoda K (2015) Practical Integration-Free Episomal Methods for Generating Human Induced Pluripotent Stem Cells Current Protocols in Human Genetics. Curr Protoc Hum Genet 87. https://doi.org/10.1002/0471142905.hg2102s87
Okita K, Yamakawa T, Matsumura Y, Sato Y, Amano N, Watanabe A, Goshima N, Yamanaka S (2013) An efficient nonviral method to generate integration-free human-induced pluripotent stem cells from cord blood and peripheral blood cells. Stem Cells 31:458–466. https://doi.org/10.1002/stem.1293
Swaidan NT, Salloum-Asfar S, Palangi F, Errafii K, Soliman NH, Aboughalia AT, Wali AHS, Abdulla SA, Emara MM (2020) Identification of potential transcription factors that enhance human iPSC generation. Sci Rep 10:21950. https://doi.org/10.1038/s41598-020-78932-9
Ohlemacher SK, Sridhar A, Xiao Y, Hochstetler AE, Sarfarazi M, Cummins TR, Meyer JS (2016) Stepwise differentiation of retinal ganglion cells from human pluripotent stem cells enables analysis of glaucomatous neurodegeneration: hPSC-derived RGCs and Glaucoma. Stem Cells 34:1553–1562. https://doi.org/10.1002/stem.2356
Ohlemacher SK, Iglesias CL, Sridhar A, Gamm DM, Meyer JS (2015) Generation of highly enriched populations of optic vesicle−like retinal cells from human pluripotent stem cells. Curr Protoc Stem Cell Biol 32. https://doi.org/10.1002/9780470151808.sc01h08s32
Acknowledgments
This work is supported by grants from National Institute of Health (R01 GM109434 and GM130129), and YB is also supported by William and Ella Owens Medical Research Foundation.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2021 Springer Science+Business Media, LLC
About this protocol
Cite this protocol
Singh, P. et al. (2021). Creating Cell Model 2.0 Using Patient Samples Carrying a Pathogenic Mitochondrial DNA Mutation: iPSC Approach for LHON. In: Turksen, K. (eds) Induced Pluripotent Stem Cells and Human Disease. Methods in Molecular Biology, vol 2549. Humana, New York, NY. https://doi.org/10.1007/7651_2021_384
Download citation
DOI: https://doi.org/10.1007/7651_2021_384
Published:
Publisher Name: Humana, New York, NY
Print ISBN: 978-1-0716-2584-2
Online ISBN: 978-1-0716-2585-9
eBook Packages: Springer Protocols