Human induced pluripotent stem cells generated from a patient with idiopathic basal ganglia calcification

Idiopathic basal ganglia calcification (IBGC) is a rare neurodegenerative disease, characterized by abnormal calcium deposits in basal ganglia of the brain. The affected individuals exhibit movement disorders, and progressive deterioration of cognitive and psychiatric ability. The genetic cause of the disease is mutation in one of several different genes, SLC20A2, PDGFB, PDGFRB, XPR1 or MYORG, which inheritably or sporadically occurs. Here we generated an induced pluripotent stem cell (iPSC) line from an IBGC patient, which is likely be a powerful tool for revealing the pathomechanisms and exploring potential therapeutic candidates of IBGC

Simple derivation of skeletal muscle from human pluripotent stem cells using temperature‐sensitive Sendai virus vector

温度感受性センダイウイルスベクターを用いて ヒトES細胞／iPS細胞から骨格筋細胞を簡便に作製する技術開発 --神経筋疾患病態モデル構築と創薬研究への利用--. 京都大学プレスリリース. 2021-09-13.Human pluripotent stem cells have the potential to differentiate into various cell types including skeletal muscles (SkM), and they are applied to regenerative medicine or in vitro modelling for intractable diseases. A simple differentiation method is required for SkM cells to accelerate neuromuscular disease studies. Here, we established a simple method to convert human pluripotent stem cells into SkM cells by using temperature-sensitive Sendai virus (SeV) vector encoding myoblast determination protein 1 (SeV-Myod1), a myogenic master transcription factor. SeV-Myod1 treatment converted human embryonic stem cells (ESCs) into SkM cells, which expressed SkM markers including myosin heavy chain (MHC). We then removed the SeV vector by temporal treatment at a high temperature of 38℃, which also accelerated mesodermal differentiation, and found that SkM cells exhibited fibre-like morphology. Finally, after removal of the residual human ESCs by pluripotent stem cell-targeting delivery of cytotoxic compound, we generated SkM cells with 80% MHC positivity and responsiveness to electrical stimulation. This simple method for myogenic differentiation was applicable to human-induced pluripotent stem cells and will be beneficial for investigations of disease mechanisms and drug discovery in the future

Establishment of induced pluripotent stem cells from schizophrenia discordant fraternal twins

Schizophrenia (SCZ) is one of the major psychiatric disorders. The genetic factor is certainly influential in the onset of the disease but is not decisive. There is no identified molecular/cellular marker of the disease, and the pathomechanism is still unknown. In this study, we generated human induced pluripotent stem cells (iPSCs) derived from SCZ-discordant fraternal twins, and they could contribute to elucidation of the pathomechanism of SCZ

Generation of four induced pluripotent stem cell lines (FHUi003-A, FHUi003-B, FHUi004-A and FHUi004-B) from two affected individuals of a familial neurohypophyseal diabetes insipidus family

Four disease-specific induced pluripotent stem cell (iPSC) lines were respectively derived from peripheral blood mononuclear cells of two affected individuals in a family affected by familial neurohypophyseal diabetes insipidus carrying the c.314G>C mutation. The expression of pluripotency markers (NANOG, OCT4, and SOX2), maintenance of a normal karyotype, absence of episomal vectors used for iPSC generation, and presence of the original pathogenic mutation were confirmed for each iPSC line. The ability to differentiate into three germ layers was confirmed by a teratoma formation assay. These iPSC lines can help in disease recapitulation in vitro using organoids and elucidation of disease mechanisms

Recombinant human thrombomodulin inhibits neutrophil extracellular trap formation in vitro

The aim of this study was to investigate the effects of recombinant human-soluble thrombomodulin (rTM) on lipopolysaccharide (LPS)-induced, platelet-dependent neutrophil extracellular trap (NET) formation (NETosis). Human peripheral blood neutrophils and platelets were co-incubated with or without LPS (0.2 μg/ml) in the presence and absence of rTM (2 μg/ml). NETosis was confirmed by immunostaining and confocal microscopy. In the absence of platelets, LPS did not induce NETosis in the neutrophils. NETosis, however, was induced by LPS when neutrophils were co-cultured with platelets (64 % of neutrophils). Notably, rTM was able to fully inhibit NETosis in neutrophils cultured with platelets and in the presence of LPS. rTM did not induce NETosis in this co-culture system (p < 0.01 versus LPS in the absence of rTM). These results show that rTM can suppress LPS-induced platelet-dependent NETosis in vitro

Generation of a human induced pluripotent stem cell line, BRCi009-A, derived from a patient with glycogen storage disease type 1a

Glycogen storage disease type 1a (GSD1a) is an autosomal recessive disorder caused by mutations of the glucose-6-phosphatase (G6PC) gene. Mutations of the G6PC gene lead to excessive accumulation of glycogen in the liver, kidney, and intestinal mucosa due to the deficiency of microsomal glucose-6-phosphatase.Human induced pluripotent stem cells (iPSCs) enable the production of patient-derived hepatocytes in culture and are therefore a promising tool for modeling GSD1a. Here, we report the establishment of human iPSCs from a GSD1a patient carrying a G6PC mutation (c.648G > T; p.Leu216 = )

Die Rolle der HECT-Typ Ubiquitin Ligasen WWP1 und WWP2 bei der Entwicklung und der Funktion von Nervenzellen

Neuste Untersuchen zeigen, dass die posttranslationale Modifikation von Proteinen durch das Ubiquitin-System ein bedeutender Regulationsmechanismus in der neuronalen Entwicklung und der Funktion von neuronalen Systemen darstellt. Die eukaryotischen “WW-Domänen enthaltenen Proteine 1 und 2” (WWP1 und WWP2) sind evolutionär konservierte E3 Ubiquitin Ligasen die eine HECT-Domäne aufweisen. Das zu WWP1/2 orthologe Protein in C. elegans spielt eine zentrale Rolle in der Funktion des Nervensystems z.B. der axonalen Wegfindung und der synaptischen Signalübertragung. Die Funktion von WWP1/2 im Gehirn von Säugetieren wurde jedoch noch nicht analysiert und auch die entsprechenden Substrate dieser Ubiquitin Ligasen sind nicht bekannt. In der vorliegenden Studie deckte ich eine neuartige Funktion von WWP1/2 in den sich entwickelnden Säugetiergehirnen auf. Analysen von Gehirnen von Mäusen, mit einer spezifischen und begrenzten Ausschaltung der Gene für WWP1/2 im Gehirn, in vivo und in vitro zeigten, dass WWP1 und 2 entscheidend für die Etablierung der Polarität von Nervenzellen sind. Die Ausbildung von einzelnen Axonen und mehreren Dendriten ist ein grundlegender Schritt während der Entwicklung einzelner Nervenzellen und neuronaler Systeme. Weiterhin konnte ich zeigen, dass die SAD-A Kinase, ein wichtiges Protein für die Entwicklung neuronaler Polarität in der Großhirnrinde, von WWP1/2 ubiquitiniert wird. Demzufolge schlussfolgere ich, dass WWP1/2 eine Rolle bei der Ausbildung einer polarisierten Zellmorphologie von Nervenzellen, über die Regulation des SAD-A Kinase Signalweges, spielen könnte. Weiterhin suchte ich nach Proteinen, welche mit WWP1/2 im Gehirn interagieren. Dabei konnte ich zeigen, dass die synaptischen Gerüstproteine Liprin-α3 und Shank1a spezifisch von WWP1/2 ubiquitiniert werden. Diese Ergebnisse stimmen mit der Lokalisation von WWP1/2 in den Synapsen von reifen Neuronen überein und deuten darauf hin, dass WWP1 und 2 an der synaptischen Signalübertragung und Synaptogenese beteiligt sind. Zusammenfassend zeigen meine Ergebnisse, dass WWP1 und 2 vielfache Funktionen in der Entwicklung und der Funktion von Synapsen im Säugetiergehirn haben

Modeling Neurological Disorders with Human Pluripotent Stem Cell-Derived Astrocytes

Astrocytes play vital roles in neurological disorders. The use of human induced pluripotent stem cell (iPSC)-derived astrocytes provides a chance to explore the contributions of astrocytes in human diseases. Here we review human iPSC-based models for neurological disorders associated with human astrocytes and discuss the points of each model