IDH1R132H in Neural Stem Cells: Differentiation Impaired by Increased Apoptosis

Autor: Marta Winiecka-Klimek, Sylwester Piaskowski, Barbara Krynska, Kamila Rosiak, Piotr Rieske, Dawid P. Grzela, Maciej Smolarz, Ewelina Stoczynska-Fidelus, Wojciech J. Stec, Joanna Peciak
Rok vydání: 2016
Předmět:
0301 basic medicine
Cellular differentiation
lcsh:Medicine
Gene Expression
Apoptosis
Embryoid body
0302 clinical medicine
Neural Stem Cells
Animal Cells
Medicine and Health Sciences
lcsh:Science
Induced pluripotent stem cell
Neurological Tumors
Cells
Cultured
Cultured Tumor Cells
Neurons
Multidisciplinary
Cell Death
Caspase 3
Stem Cells
Neurogenesis
Cell Differentiation
Glioma
Isocitrate Dehydrogenase
Neural stem cell
Cell biology
Isocitrate dehydrogenase
Oncology
Neurology
Cell Processes
030220 oncology & carcinogenesis
Biological Cultures
Cellular Types
Neuronal Differentiation
Research Article
IDH1
Induced Pluripotent Stem Cells
Biology
Research and Analysis Methods
03 medical and health sciences
Developmental Neuroscience
Genetics
Humans
Cell Lineage
Epigenetics
Differentiated Tumors
Embryoid Bodies
lcsh:R
Biology and Life Sciences
Cancers and Neoplasms
Cell Biology
Cell Cultures
Glioma Cells
030104 developmental biology
Cellular Neuroscience
Astrocytes
lcsh:Q
Biomarkers
Developmental Biology
Neuroscience
Zdroj: PLoS ONE
PLoS ONE, Vol 11, Iss 5, p e0154726 (2016)
ISSN: 1932-6203
Popis: BACKGROUND:The high frequency of mutations in the isocitrate dehydrogenase 1 (IDH1) gene in diffuse gliomas indicates its importance in the process of gliomagenesis. These mutations result in loss of the normal function and acquisition of the neomorphic activity converting α-ketoglutarate to 2-hydroxyglutarate. This potential oncometabolite may induce the epigenetic changes, resulting in the deregulated expression of numerous genes, including those related to the differentiation process or cell survivability. METHODS:Neural stem cells were derived from human induced pluripotent stem cells following embryoid body formation. Neural stem cells transduced with mutant IDH1R132H, empty vector, non-transduced and overexpressing IDH1WT controls were differentiated into astrocytes and neurons in culture. The neuronal and astrocytic differentiation was determined by morphology and expression of lineage specific markers (MAP2, Synapsin I and GFAP) as determined by real-time PCR and immunocytochemical staining. Apoptosis was evaluated by real-time observation of Caspase-3 activation and measurement of PARP cleavage by Western Blot. RESULTS:Compared with control groups, cells expressing IDH1R132H retained an undifferentiated state and lacked morphological changes following stimulated differentiation. The significant inhibitory effect of IDH1R132H on neuronal and astrocytic differentiation was confirmed by immunocytochemical staining for markers of neural stem cells. Additionally, real-time PCR indicated suppressed expression of lineage markers. High percentage of apoptotic cells was detected within IDH1R132H-positive neural stem cells population and their derivatives, if compared to normal neural stem cells and their derivatives. The analysis of PARP and Caspase-3 activity confirmed apoptosis sensitivity in mutant protein-expressing neural cells. CONCLUSIONS:Our study demonstrates that expression of IDH1R132H increases apoptosis susceptibility of neural stem cells and their derivatives. Robust apoptosis causes differentiation deficiency of IDH1R132H-expressing cells.
Databáze: OpenAIRE
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