Abstract
Leigh syndrome (LS) is a rare, inherited neurometabolic disorder that presents with bilateral brain lesions caused by defects in the mitochondrial respiratory chain and associated nuclear-encoded proteins. We generated human induced pluripotent stem cells (iPSCs) from three LS patient-derived fibroblast lines. Using wholeexome and mitochondrial sequencing, we identified unreported mutations in pyruvate dehydrogenase (GM0372, PDH; GM13411, MT-ATP6/PDH) and dihydrolipoyl dehydrogenase (GM01503, DLD). These LS patient-derived iPSC lines were viable and capable of differentiating into progenitor populations, but we identified several abnormalities in three-dimensional differentiation models of brain development. LS patient-derived cerebral organoids showed defects in neural epithelial bud generation, size and cortical architecture at 100 days. The double mutant MT-ATP6/PDH line produced organoid neural precursor cells with abnormal mitochondrial morphology, characterized by fragmentation and disorganization, and showed an increased generation of astrocytes. These studies aim to provide a comprehensive phenotypic characterization of available patient-derived cell lines that can be used to study Leigh syndrome.
| Original language | English (US) |
|---|---|
| Article number | dev199914 |
| Journal | Development (Cambridge) |
| Volume | 149 |
| Issue number | 20 |
| DOIs | |
| State | Published - Oct 2022 |
Funding
Funding was provided by National Institutes of Health grants 1R35GM128915-01, 1R21 CA227483-01A1 and 1RF1MH123971-01 to V.G., and 1F99 NS125829-01 to G.L.R., and the Precision Medicine and Mental Health Initiative sponsored by the Vanderbilt Brain Institute at Vanderbilt University to V.G. Open Access funding provided by National Institutes of Health. Deposited in PMC for immediate release. We thank Dr Nicholas Mignemi (Vanderbilt Nikon Center for Excellence) for his technical support with image acquisition and processing. We thank Stellan Riffle for technical help and all the members of the Gama Laboratory for helpful discussions and comments on the manuscript. Image acquisition and analysis were performed in part through the use of the Nikon Center of Excellence within the Vanderbilt Cell Imaging Shared Resource (supported by National Institutes of Health (NIH) grants CA68485, DK20593, DK58404, DK59637 and EY08126), Vanderbilt University Medical Center’s Translational Pathology Shared Resource supported by National Cancer Institute (NCI)/NIH Cancer Center Support Grant 2P30 CA068485-14, and the Vanderbilt Mouse Metabolic Phenotyping Center Grant 5U24DK059637-13. Whole-exome sequencing and mitochondrial sequencing results were analyzed by Creative Solutions. Metabolite measurements were performed by the Northwestern University RHLCCC Metabolomics Core (Dr Peng Gao) and were supported by NIH grants NIH2PO1HL071643-11A1, NIH1R35CA197532-01 and NIH1PO1AG049665-01 to N.S.C., NIH/NCI grant T32CA09560 to G.S.M. and Northwestern University Pulmonary and Critical Care Department’s Cugell Predoctoral Fellowship to R.P.C.
Keywords
- Brain organoids
- Glycolysis
- Leigh syndrome
- Mitochondria
- Neural precursor cells
- Neural rosettes
- Oxidative phosphorylation
- Stem cells
ASJC Scopus subject areas
- Molecular Biology
- Developmental Biology
