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The development of an in vitro cerebral organoid model for investigating the pathomolecular mechanisms associated with the Central Nervous System involvement in Mitochondrial Neurogastrointestinal Encephalomyopathy

Pacitti, D; Bax, BE (2018) The development of an in vitro cerebral organoid model for investigating the pathomolecular mechanisms associated with the Central Nervous System involvement in Mitochondrial Neurogastrointestinal Encephalomyopathy. Nucleosides, Nucleotides and Nucleic Acids, 37 (11). pp. 603-617. ISSN 1525-7770 https://doi.org/10.1080/15257770.2018.1492139
SGUL Authors: Bax, Bridget Elizabeth

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Abstract

Mitochondrial neurogastrointestinal encephalomyopathy (MNGIE) is a rare disorder caused by mutations in the thymidine phosphorylase gene (TYMP), leading to secondary aberrations to the mitochondrial genome. The disease is characterised by gastrointestinal dysmotility, sensorimotor peripheral neuropathy and leukoencephalopathy. The understanding of the molecular mechanisms that underlie the central nervous system (CNS) is hindered by the lack of a representative disease model; to address this we have developed an in vitro 3-D cerebral organoid of MNGIE. Induced pluripotent stem cells (iPSCs) generated from peripheral blood mononuclear cells (PBMC) of a healthy control and a patient with MNGIE were characterised to ascertain bona fide pluripotency through the evaluation of pluripotency markers and the differentiation to the germ layers. iPSC lines were differentiated into cerebral organoids. Thymidine phosphorylase expression in PBMCs, iPSCs and Day 92 organoids was evaluated by immunoblotting and intact organoids were sampled for histological evaluation of neural markers. iPSCs demonstrated the expression of pluripotency markers SOX2 and TRA1-60 and the plasticity to differentiate into the germ layers. Cerebral organoids stained positive for the neural markers GFAP, O4, Tuj1, Nestin, SOX2 and MBP. Consistent with the disease phenotypes, MNGIE cells did not display thymidine phosphorylase expression whereas control PBMCs and Day 92 organoids did. Remarkably, control iPSCs did not stain positive for thymidine phosphorylase. We have established for the first time a MNGIE iPSC line and cerebral organoid model, which exhibited the expression of cells relevant to the study of the disease, such as neural stem cells, astrocytes and myelinating oligodendrocytes.

Item Type: Article
Additional Information: This is an Accepted Manuscript of an article published by Taylor & Francis in Nucleosides, Nucleotides and Nucleic Acids on 27/12/18, available online: http://www.tandfonline.com/10.1080/15257770.2018.1492139.
SGUL Research Institute / Research Centre: Academic Structure > Molecular and Clinical Sciences Research Institute (MCS)
Journal or Publication Title: Nucleosides, Nucleotides and Nucleic Acids
ISSN: 1525-7770
Dates:
DateEvent
27 December 2018Published
14 June 2018Accepted
Publisher License: Publisher's own licence
Projects:
Project IDFunderFunder ID
UNSPECIFIEDPurine Metabolic Patients AssociationUNSPECIFIED
URI: https://openaccess.sgul.ac.uk/id/eprint/110022
Publisher's version: https://doi.org/10.1080/15257770.2018.1492139

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