Manifestation of Huntington’s disease pathology in human induced pluripotent stem cell-derived neurons

Abstract

BackgroundHuntington’s disease (HD) is an incurable hereditary neurodegenerative disorder, which manifests itself as a loss of GABAergic medium spiny (GABA MS) neurons in the striatum and caused by an expansion of the CAG repeat in exon 1 of the huntingtin gene. There is no cure for HD, existing pharmaceutical can only relieve its symptoms.ResultsHere, induced pluripotent stem cells were established from patients with low CAG repeat expansion in the huntingtin gene, and were then efficiently differentiated into GABA MS-like neurons (GMSLNs) under defined culture conditions. The generated HD GMSLNs recapitulated disease pathology in vitro, as evidenced by mutant huntingtin protein aggregation, increased number of lysosomes/autophagosomes, nuclear indentations, and enhanced neuronal death during cell aging. Moreover, store-operated channel (SOC) currents were detected in the differentiated neurons, and enhanced calcium entry was reproducibly demonstrated in all HD GMSLNs genotypes. Additionally, the quinazoline derivative, EVP4593, reduced the number of lysosomes/autophagosomes and SOC currents in HD GMSLNs and exerted neuroprotective effects during cell aging.ConclusionsOur data is the first to demonstrate the direct link of nuclear morphology and SOC calcium deregulation to mutant huntingtin protein expression in iPSCs-derived neurons with disease-mimetic hallmarks, providing a valuable tool for identification of candidate anti-HD drugs. Our experiments demonstrated that EVP4593 may be a promising anti-HD drug.Electronic supplementary materialThe online version of this article (doi:10.1186/s13024-016-0092-5) contains supplementary material, which is available to authorized users.

Huntington’s disease modeling on HEK293 cell line

“…Не обнаружив внутриклеточных агрегатов в мутантных клетках, мы решили индуцировать их формирование с помощью ингибитора протеасом MG132, так как ранее в исследованиях его уже использовали в этих целях (Jeon et al, 2012;Nekrasov et al, 2016). В ходе этого эксперимента показано, что белковые агрегаты также не образуются, но клетки, несущие мутацию, более чувствительны к добавлению ингибитора.…”

A Platform for Studying Molecular and Cellular Mechanisms of Parkinson’s Disease Based on Human Induced Pluripotent Stem Cells

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Genetic basis of neurological disorder with human induced pluripotent stem cells

“…According to Nekrasov et al, 2016 established iPSCs from the patient who has low expansion repeats of CAG in HTT gene and it is differentiated into GABA MS -like neurons (GMSLNs) under suitable culture. This HD GMSLNs was analysed for diseased pathology with the help of information such as mutant huntingtin protein aggregation, increased number of lysosomes/autophagosomes, nuclear indentations, and enhanced neuronal death during cell aging.…”