Human iPSC-derived cerebral organoids model features of Leigh syndrome and reveal abnormal corticogenesis

Alejandra I. Romero-Morales; Gabriella L. Robertson; Anuj Rastogi; Megan L. Rasmussen; Hoor Temuri; Gregory Scott McElroy; Ram Prosad Chakrabarty; Lawrence Hsu; Paula M. Almonacid; Bryan A. Millis; Navdeep S. Chandel; Jean Philippe Cartailler; Vivian Gama

doi:10.1242/dev.199914/2151195/dev199914

Human iPSC-derived cerebral organoids model features of Leigh syndrome and reveal abnormal corticogenesis

Alejandra I. Romero-Morales, Gabriella L. Robertson, Anuj Rastogi, Megan L. Rasmussen, Hoor Temuri, Gregory Scott McElroy, Ram Prosad Chakrabarty, Lawrence Hsu, Paula M. Almonacid, Bryan A. Millis, Navdeep S. Chandel, Jean Philippe Cartailler, Vivian Gama^*

^*Corresponding author for this work

Medicine, Pulmonary Division

Research output: Contribution to journal › Article › peer-review

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	https://doi.org/10.1242/dev.199914/2151195/dev199914
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

Access to Document

10.1242/dev.199914/2151195/dev199914

Cite this

Romero-Morales, A. I., Robertson, G. L., Rastogi, A., Rasmussen, M. L., Temuri, H., McElroy, G. S., Chakrabarty, R. P., Hsu, L., Almonacid, P. M., Millis, B. A., Chandel, N. S., Cartailler, J. P., & Gama, V. (2022). Human iPSC-derived cerebral organoids model features of Leigh syndrome and reveal abnormal corticogenesis. Development (Cambridge), 149(20), Article dev199914. https://doi.org/10.1242/dev.199914/2151195/dev199914

@article{e2d43ebbd0c847f291fa3ff988d364ef,

title = "Human iPSC-derived cerebral organoids model features of Leigh syndrome and reveal abnormal corticogenesis",

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.",

keywords = "Brain organoids, Glycolysis, Leigh syndrome, Mitochondria, Neural precursor cells, Neural rosettes, Oxidative phosphorylation, Stem cells",

author = "Romero-Morales, {Alejandra I.} and Robertson, {Gabriella L.} and Anuj Rastogi and Rasmussen, {Megan L.} and Hoor Temuri and McElroy, {Gregory Scott} and Chakrabarty, {Ram Prosad} and Lawrence Hsu and Almonacid, {Paula M.} and Millis, {Bryan A.} and Chandel, {Navdeep S.} and Cartailler, {Jean Philippe} and Vivian Gama",

note = "Publisher Copyright: {\textcopyright} 2022. Published by The Company of Biologists Ltd.",

year = "2022",

month = oct,

doi = "10.1242/dev.199914/2151195/dev199914",

language = "English (US)",

volume = "149",

journal = "Development (Cambridge)",

issn = "0950-1991",

publisher = "Company of Biologists Ltd",

number = "20",

}

Romero-Morales, AI, Robertson, GL, Rastogi, A, Rasmussen, ML, Temuri, H, McElroy, GS, Chakrabarty, RP, Hsu, L, Almonacid, PM, Millis, BA, Chandel, NS, Cartailler, JP & Gama, V 2022, 'Human iPSC-derived cerebral organoids model features of Leigh syndrome and reveal abnormal corticogenesis', Development (Cambridge), vol. 149, no. 20, dev199914. https://doi.org/10.1242/dev.199914/2151195/dev199914

TY - JOUR

T1 - Human iPSC-derived cerebral organoids model features of Leigh syndrome and reveal abnormal corticogenesis

AU - Romero-Morales, Alejandra I.

AU - Robertson, Gabriella L.

AU - Rastogi, Anuj

AU - Rasmussen, Megan L.

AU - Temuri, Hoor

AU - McElroy, Gregory Scott

AU - Chakrabarty, Ram Prosad

AU - Hsu, Lawrence

AU - Almonacid, Paula M.

AU - Millis, Bryan A.

AU - Chandel, Navdeep S.

AU - Cartailler, Jean Philippe

AU - Gama, Vivian

PY - 2022/10

Y1 - 2022/10

N2 - 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.

AB - 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.

KW - Brain organoids

KW - Glycolysis

KW - Leigh syndrome

KW - Mitochondria

KW - Neural precursor cells

KW - Neural rosettes

KW - Oxidative phosphorylation

KW - Stem cells

UR - http://www.scopus.com/inward/record.url?scp=85140376661&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85140376661&partnerID=8YFLogxK

U2 - 10.1242/dev.199914/2151195/dev199914

DO - 10.1242/dev.199914/2151195/dev199914

M3 - Article

C2 - 35792828

AN - SCOPUS:85140376661

SN - 0950-1991

VL - 149

JO - Development (Cambridge)

JF - Development (Cambridge)

IS - 20

M1 - dev199914

ER -

Human iPSC-derived cerebral organoids model features of Leigh syndrome and reveal abnormal corticogenesis

Abstract

Funding

Keywords

ASJC Scopus subject areas

Access to Document

Other files and links

Fingerprint

Cite this