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.
Highlights
Huntington’s disease (HD) is an incurable hereditary neurodegenerative disorder, which manifests itself as a loss of GABAergic medium spiny (GABA GABAergic Medium Spiny (MS)) neurons in the striatum and caused by an expansion of the CAG repeat in exon 1 of the huntingtin gene
Recapitulation of disease pathology phenotype in the HD neuron model To elucidate specific differences in the ability of pathological vs. normal pluripotent stem cells (PSCs) to differentiate into neurons, we examined proliferation rates, neural progenitor cells (NPCs) forming capacity, and the relative amount of DARPP-32 positive neurons in GABAergic medium spiny (GABA MS)-like neurons (GMSLNs) cultures generated from the HD and the WT PSC lines
We found that allele-specific knockdown of mutant huntingtin expression in HD GMSLNs using antisense oligonucleotide (Additional file 1: Figure S3C) decreased amplitude of abnormal store-operated channel (SOC)-mediated calcium entry from 4.62 ± 0.61 pA/ pF in cells transfected with control LNA(S) to 2.29 ± 0.38 pA/pF in cells transfected with LNA(T) targeting mutant HTT (mHTT) (Fig. 3d, f)
Summary
Huntington’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. Huntington’s disease (HD) is an incurable autosomal dominant hereditary neurodegenerative disorder that typically manifests between 35–55 years of age. HD is caused by an expansion of cytosine-adenineguanine (CAG) repeats in the huntingtin gene (HTT) that leads to a pathological elongation of polyglutamine repeats in the huntingtin protein (HTT). The HD phenotype develops when the number of trinucleotide repeats in the HTT gene exceeds 36. The HTT protein normally interacts with hundreds of other proteins, and probably has multiple biological functions [2]. While wild-type HTT (wtHTT) and mutant HTT (mHTT)
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