Use of induced pluripotent stem cells to investigate the effects of purine nucleoside phosphorylase deficiency on neuronal development

Background: Inherited defects in the function of the purine nucleoside phosphorylase (PNP) enzyme can cause severe T cell immune deficiency and early death from infection, autoimmunity, or malignancy. In addition, more than 50% of patients suffer diverse non-infectious neurological complications. However the cause for the neurological abnormalities are not known. Objectives: Differentiate induced pluripotent stem cells (iPSC) from PNP-deficient patients into neuronal cells to better understand the effects of impaired purine metabolism on neuronal development. Methods: Sendai virus was used to generate pluripotent stem cells from PNP-deficient and healthy control lymphoblastoid cells. Cells were differentiated into neuronal cells through the formation of embryoid bodies. Results: After demonstration of pluripotency, normal karyotype, and retention of the PNP deficiency state, iPSC were differentiated into neuronal cells. PNP-deficient neuronal cells had reduced soma and nuclei size in comparison to cells derived from healthy controls. Spontaneous apoptosis, determined by Caspase-3 expression, was increased in PNP-deficient cells. Conclusions: iPSC from PNP-deficient patients can be differentiated into neuronal cells, thereby providing an important tool to study the effects of impaired purine metabolism on neuronal development and potential treatments. Statement of novelty: We report here the first generation and use of neuronal cells derived from induced pluripotent stem cells to model human PNP deficiency, thereby providing an important tool for better understanding and management of this condition.

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Differentiation of blood T cells: Reprogramming human induced pluripotent stem cells into neuronal cells

Journal of the Chinese Medical Association ◽

10.1016/j.jcma.2015.03.007 ◽

2015 ◽

Vol 78 (6) ◽

pp. 353-359 ◽

Cited By ~ 2

Author(s):

Ping-Hsing Tsai ◽

Yun-Ching Chang ◽

Yi-Yen Lee ◽

Yu-Ling Ko ◽

Yu-Hsuan Yang ◽

...

Keyword(s):

Stem Cells ◽

T Cells ◽

Induced Pluripotent Stem Cells ◽

Pluripotent Stem Cells ◽

Neuronal Cells ◽

Induced Pluripotent

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Using induced pluripotent stem cells to investigate human neuronal phenotypes in 1q21.1 deletion and duplication syndrome

10.1101/2021.02.08.430246 ◽

2021 ◽

Author(s):

Gareth Chapman ◽

Mouhamed Alsaqati ◽

Sharna Lunn ◽

Tanya Singh ◽

Stefanie C Linden ◽

...

Keyword(s):

Stem Cells ◽

Induced Pluripotent Stem Cells ◽

Pluripotent Stem Cells ◽

Cortical Neurons ◽

Neuronal Development ◽

Motor Deficits ◽

Differentiation Potential ◽

Induced Pluripotent ◽

The Impact ◽

1Q21.1 Deletion

AbstractCopy Number Variation (CNV) at the 1q21.1 locus is associated with a range of neurodevelopmental and psychiatric disorders in humans, including abnormalities in head size and motor deficits. Yet, the functional consequences of these CNVs (both deletion and duplication) on neuronal development remain unknown. To determine the impact of CNV at the 1q21.1 locus on neuronal development, we generated induced pluripotent stem cells from individuals harbouring 1q21.1 deletion or duplication and differentiated them into functional cortical neurons. We show that neurons with 1q21.1 deletion or duplication display reciprocal phenotype with respect to proliferation, differentiation potential, neuronal maturation, synaptic density, and functional activity. Deletion of the 1q21.1 locus was also associated with an increased expression of lower cortical layer markers. This difference was conserved in the mouse model of 1q21.1 deletion, which displayed altered corticogenesis. Importantly, we show that neurons with 1q21.1 deletion and duplication are associated with differential expression of calcium channels and demonstrate that physiological deficits in neurons with 1q21.1 deletion or duplication can be pharmacologically modulated by targeting Ca2+ channel activity. These findings provide biological insight into the neuropathological mechanism underlying 1q21.1 associated brain disorder and indicate a potential target for therapeutic interventions.

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ELUCIDATING THE MOLECULAR PATHOPHYSIOLOGY OF MENTAL ILLNESS WITH OLFACTORY NEURONAL CELLS AND INDUCED PLURIPOTENT STEM CELLS

Schizophrenia Research ◽

10.1016/s0920-9964(14)70146-x ◽

2014 ◽

Vol 153 ◽

pp. S45

Author(s):

Akira Sawa ◽

Yasue Horiuchi ◽

Nicola Cascella ◽

David Schretlen ◽

Koko Ishizuka

Keyword(s):

Stem Cells ◽

Mental Illness ◽

Induced Pluripotent Stem Cells ◽

Pluripotent Stem Cells ◽

Neuronal Cells ◽

Induced Pluripotent ◽

Molecular Pathophysiology

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Using induced pluripotent stem cells to investigate human neuronal phenotypes in 1q21.1 deletion and duplication syndrome

Molecular Psychiatry ◽

10.1038/s41380-021-01182-2 ◽

2021 ◽

Author(s):

Gareth Chapman ◽

Mouhamed Alsaqati ◽

Sharna Lunn ◽

Tanya Singh ◽

Stefanie C. Linden ◽

...

Keyword(s):

Stem Cells ◽

Induced Pluripotent Stem Cells ◽

Pluripotent Stem Cells ◽

Cortical Neurons ◽

Neuronal Development ◽

Motor Deficits ◽

Differentiation Potential ◽

Induced Pluripotent ◽

The Impact ◽

1Q21.1 Deletion

AbstractCopy Number Variation (CNV) at the 1q21.1 locus is associated with a range of neurodevelopmental and psychiatric disorders in humans, including abnormalities in head size and motor deficits. Yet, the functional consequences of these CNVs (both deletion and duplication) on neuronal development remain unknown. To determine the impact of CNV at the 1q21.1 locus on neuronal development, we generated induced pluripotent stem cells from individuals harbouring 1q21.1 deletion or duplication and differentiated them into functional cortical neurons. We show that neurons with 1q21.1 deletion or duplication display reciprocal phenotype with respect to proliferation, differentiation potential, neuronal maturation, synaptic density and functional activity. Deletion of the 1q21.1 locus was also associated with an increased expression of lower cortical layer markers. This difference was conserved in the mouse model of 1q21.1 deletion, which displayed altered corticogenesis. Importantly, we show that neurons with 1q21.1 deletion and duplication are associated with differential expression of calcium channels and demonstrate that physiological deficits in neurons with 1q21.1 deletion or duplication can be pharmacologically modulated by targeting Ca2+ channel activity. These findings provide biological insight into the neuropathological mechanism underlying 1q21.1 associated brain disorder and indicate a potential target for therapeutic interventions.

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Selective linkage of mitochondrial enzymes to intracellular calcium stores differs between human induced pluripotent stem cells, neural stem cells and neurons

10.1101/2020.06.20.162040 ◽

2020 ◽

Author(s):

Huanlian Chen ◽

Ankita Thakkar ◽

Abigail C. Cross ◽

Hui Xu ◽

Aiqun Li ◽

...

Keyword(s):

Stem Cells ◽

Neural Stem Cells ◽

Induced Pluripotent Stem Cells ◽

Pluripotent Stem Cells ◽

Neuronal Development ◽

Calcium Stores ◽

Mitochondrial Enzymes ◽

Induced Pluripotent ◽

Er Calcium ◽

Dehydrogenase Complex

AbstractThe coupling of the endoplasmic reticulum (ER) with mitochondria modulates neuronal calcium signaling. Whether this link changes with neuronal development is unknown. The current study first determined whether ER calcium stores are similar during development of human neurons, and then tested if the ER/mitochondrial coupling varied with development. The release of ER calcium to the cytosol by the IP3 agonist bradykinin was determined in human induced-pluripotent stem cells (iPSC), neural stem cells (NSC) and neurons. The concentration dependence for the release of ER calcium was similar at different stages of development. Metabolism changes dramatically with development. Glycolysis is the main energy source in iPSC and NSC whereas mitochondrial metabolism is more prominent in neurons. To test whether the coupling of mitochondria and ER changed with development, bombesin or bradykinin releasable calcium stores (BRCS) were monitored after inhibiting either of two key mitochondrial enzyme complexes: the alpha-ketoglutarate dehydrogenase complex (KGDHC) or the pyruvate dehydrogenase complex (PDHC). Inhibition of KGDHC did not alter BRCS in either iPSC or NSC. Inhibition of PDHC in neurons diminished BRCS whereas decreased KGDHC activity exaggerated BRCS. The latter finding may help understand the pathology of Alzheimer’s disease (AD). BRCS is exaggerated in cells from AD patients and KGDHC is reduced in brains of patients with AD. In summary, a prominent ER/mitochondrial link in neurons is associated with selective mitochondrial enzymes. The ER/mitochondrial link changes with human neuronal development and plausibly links ER calcium changes to AD.

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Modeling the neuropsychiatric manifestations of Lowe syndrome using induced pluripotent stem cells: defective F-actin polymerization and WAVE-1 expression in neuronal cells

Molecular Autism ◽

10.1186/s13229-018-0227-3 ◽

2018 ◽

Vol 9 (1) ◽

Cited By ~ 9

Author(s):

Jesse Barnes ◽

Franklin Salas ◽

Ryan Mokhtari ◽

Hedwig Dolstra ◽

Erika Pedrosa ◽

...

Keyword(s):

Stem Cells ◽

Induced Pluripotent Stem Cells ◽

Pluripotent Stem Cells ◽

Actin Polymerization ◽

Neuronal Cells ◽

Lowe Syndrome ◽

Neuropsychiatric Manifestations ◽

Induced Pluripotent

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Glycoproteomic analysis of the changes in protein N-glycosylation during neuronal differentiation in human-induced pluripotent stem cells and derived neuronal cells

Scientific Reports ◽

10.1038/s41598-021-90102-z ◽

2021 ◽

Vol 11 (1) ◽

Author(s):

Kazumasa Kimura ◽

Takumi Koizumi ◽

Takaya Urasawa ◽

Yuki Ohta ◽

Daisuke Takakura ◽

...

Keyword(s):

Stem Cells ◽

Induced Pluripotent Stem Cells ◽

Neuronal Differentiation ◽

Pluripotent Stem Cells ◽

Neuronal Cells ◽

Neural Regeneration ◽

Post Translational Modification ◽

Integrated Method ◽

Induced Pluripotent

AbstractN-glycosylation of glycoproteins, a major post-translational modification, plays a crucial role in various biological phenomena. In central nervous systems, N-glycosylation is thought to be associated with differentiation and regeneration; however, the state and role of N-glycosylation in neuronal differentiation remain unclear. Here, we conducted sequential LC/MS/MS analyses of tryptic digest, enriched glycopeptides, and deglycosylated peptides of proteins derived from human-induced pluripotent stem cells (iPSCs) and iPSC-derived neuronal cells, which were used as a model of neuronal differentiation. We demonstrate that the production profiles of many glycoproteins and their glycoforms were altered during neuronal differentiation. Particularly, the levels of glycoproteins modified with an N-glycan, consisting of five N-acetylhexosamines, three hexoses, and a fucose (HN5H3F), increased in dopaminergic neuron-rich cells (DAs). The N-glycan was deduced to be a fucosylated and bisected biantennary glycan based on product ion spectra. Interestingly, the HN5H3F-modified proteins were predicted to be functionally involved in neural cell adhesion, axon guidance, and the semaphorin-plexin signaling pathway, and protein modifications were site-selective and DA-selective regardless of protein production levels. Our integrated method for glycoproteome analysis and resultant profiles of glycoproteins and their glycoforms provide valuable information for further understanding the role of N-glycosylation in neuronal differentiation and neural regeneration.

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Neuronal cells derived from human induced pluripotent stem cells as a functional tool of melanocortin system

Neuropeptides ◽

10.1016/j.npep.2017.04.004 ◽

2017 ◽

Vol 65 ◽

pp. 10-20

Author(s):

Nobuko Yamada-Goto ◽

Yukari Ochi ◽

Goro Katsuura ◽

Yui Yamashita ◽

Ken Ebihara ◽

...

Keyword(s):

Stem Cells ◽

Induced Pluripotent Stem Cells ◽

Pluripotent Stem Cells ◽

Neuronal Cells ◽

Melanocortin System ◽

Induced Pluripotent

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Neurobiology meets genomic science: The promise of human-induced pluripotent stem cells

Development and Psychopathology ◽

10.1017/s095457941200082x ◽

2012 ◽

Vol 24 (4) ◽

pp. 1443-1451 ◽

Cited By ~ 3

Author(s):

Hanna E. Stevens ◽

Jessica Mariani ◽

Gianfilippo Coppola ◽

Flora M. Vaccarino

Keyword(s):

Stem Cells ◽

Induced Pluripotent Stem Cells ◽

Pluripotent Stem Cells ◽

Cell Biology ◽

Neuronal Cells ◽

Induced Pluripotent Stem Cell ◽

Messenger Rnas ◽

Induced Pluripotent ◽

Developing Neurons

AbstractThe recent introduction of the induced pluripotent stem cell technology has made possible the derivation of neuronal cells from somatic cells obtained from human individuals. This in turn has opened new areas of investigation that can potentially bridge the gap between neuroscience and psychopathology. For the first time we can study the cell biology and genetics of neurons derived from any individual. Furthermore, by recapitulating in vitro the developmental steps whereby stem cells give rise to neuronal cells, we can now hope to understand factors that control typical and atypical development. We can begin to explore how human genes and their variants are transcribed into messenger RNAs within developing neurons and how these gene transcripts control the biology of developing cells. Thus, human-induced pluripotent stem cells have the potential to uncover not only what aspects of development are uniquely human but also variations in the series of events necessary for normal human brain development that predispose to psychopathology.

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