Methotrexate and Valproic Acid Affect Early Neurogenesis of Human Amniotic Fluid Stem Cells from Myelomeningocele

Sahakyan, Vardine; Pozzo, Enrico; Duelen, Robin; Deprest, Jan; Sampaolesi, Maurilio

doi:10.1155/2017/6101609

Cited by 9 publications

(6 citation statements)

References 39 publications

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“…Previous studies also showed that histone methylation modi cation could directly or indirectly activate and inhibit gene expression, thereby affecting embryonic development [30]. It was revealed that valproic acid caused the increased of H3K4me3, and decreased of H3K9me3 in the NTDs mouse model, respectively [31]. In our study, the results showed that histone methylation modi cation was signi cantly decreased in ethionine-induced NTDs embryos, which were consistent with the alteration of H3K79me3 and H3K27me3 in human NTDs samples [27,32].…”

Section: Discussionsupporting

confidence: 87%

Reduced H3K27me3 Suppresses Wnt/β-catenin Signaling by S-adenosylmethionine Deficiency in Neural Tube Development

Zhang

Wang

Cao

et al. 2021

Preprint

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Background: S-adenosylmethionine as a major methyl donor play a key role in methylation modification in vivo, and its disorder was closely related to neural tube defects. However, the underlying mechanism between SAM deficiency and NTDs remained unclear.Methods: we investigated the association between histone methylation modification and Wnt/β-catenin signaling pathway in NTDs induced by SAM deficiency. The levels of SAM and SAH were determined by enzyme linked immunosorbent assay. The expressions of H3K27me3 and Wnt/β-catenin signaling pathway specific markers were demonstrated by western blotting, reverse transcription, and quantitative PCR and immunofluorescence in ethionine induced E11.5 mouse NTDs and NSCs models. Results: we found that the incidence rate of NTDs induced by ethionine were 46.2%, post treatment of ethionine combined with SAM, the incidence rate of NTDs was reduced to 26.2%. The level of SAM was significantly decreased (P<0.05) and a reduction in the SAM/SAH ratio was observed. The SAM depletion caused the reduction of both H3K27me3 modifications and UTX activity, and inhibited the marker proteins (β-catenin, TCF-4, Axin-2, p-GSK-3β, CyclinD1, and C-myc) in Wnt/β-catenin signaling pathway (P<0.05). The differentiations of neural stem cells into neurons and oligodendrocytes were inhibited under SAM deficiency (P<0.05).Conclusions: These results indicated that the depletion of SAM led to reduced H3K27me3 modifications, prevented the activation of Wnt/β-catenin signaling pathway and NSCs differentiation, which provided an understanding of the novel function of epigenetic regulation in NTDs.

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Section: Discussionsupporting

confidence: 87%

Reduced H3K27me3 Suppresses Wnt/β-catenin Signaling by S-adenosylmethionine Deficiency in Neural Tube Development

Zhang

Wang

Cao

et al. 2021

Preprint

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“…There are several hypotheses on how VPA causes NTDs, including alterations in folate metabolism, increase in reactive oxygen species, and epigenetic dysregulation ( 30 - 34 ). For instance, the effect of VPA as a folic acid antagonist is primarily supported by a comparative analysis between the effects of methotrexate and VPA ( 35 - 37 ). This is also supported by the observation that folic acid rescued VPA-induced defects in zebrafish embryos as well as in in vitro neural tube models ( 27 , 38 ).…”

Section: Discussionmentioning

confidence: 99%

Transcriptional Signature of Valproic Acid-Induced Neural Tube Defects in Human Spinal Cord Organoids

Lee,

Shaker,

Park

et al. 2023

Int J Stem Cells

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In vertebrates, the entire central nervous system is derived from the neural tube, which is formed through a conserved early developmental morphogenetic process called neurulation. Although the perturbations in neurulation caused by genetic or environmental factors lead to neural tube defects (NTDs), the most common congenital malformation and the precise molecular pathological cascades mediating NTDs are not well understood. Recently, we have developed human spinal cord organoids (hSCOs) that recapitulate some aspects of human neurulation and observed that valproic acid (VPA) could cause neurulation defects in an organoid model. In this study, we identified and verified the significant changes in cell–cell junctional genes/proteins in VPA-treated organoids using transcriptomic and immunostaining analysis. Furthermore, VPA-treated mouse embryos exhibited impaired gene expression and NTD phenotypes, similar to those observed in the hSCO model. Collectively, our data demonstrate that hSCOs provide a valuable biological resource for dissecting the molecular pathways underlying the currently unknown human neurulation process using destructive biological analysis tools.

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“…The inhibition of neural differentiation has been shown in previous in vitro studies. 25,49 Some parallels can be drawn with human studies of MTX exposure, as fetal MTX syndrome revealed a higher number of microcephaly cases than expected, and regulation of neural crest cell differentiation could result in potential neural crest-related malformations (such as the typical facial dysmorphic features). However, this remains speculative.…”

Section: Discussionmentioning

confidence: 99%

Culture Conditions Affect Chemical-Induced Developmental Toxicity In Vitro: The Case of Folic Acid, Methionine and Methotrexate in the Neural Embryonic Stem Cell Test

et al. 2020

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In vitro tests are increasingly applied in chemical hazard assessment. Basic culture conditions may affect the outcome of in vitro tests and should be optimised to reduce false predictions. The neural embryonic stem cell test (ESTn) can predict early neurodevelopmental effects of chemicals, as it mimics the differentiation of stem cells towards the neuroectodermal lineage. Normal early neural differentiation depends crucially on folic acid (FA) and methionine (MET), both elements of the one-carbon (1C) cycle. The aim of this study was to assess the concentration-dependent influence of FA and MET on neural differentiation in the ESTn, and its consequences for assay sensitivity to methotrexate (MTX), a compound that interferes with the 1C cycle. Neural differentiation was inhibited below 0.007 mM and above 0.22 mM FA, while both stem cell viability (< 0.097 mM, > 1.52 mM) and neural differentiation (< 0.181 mM, > 1.35 mM) were affected when changing MET concentrations. A 10-day exposure to 13 nM MTX inhibited neural differentiation, especially in FA- and MET-deficient conditions. However, a 24-hour exposure to 39 nM MTX decreased neural cell and neural crest cell differentiation markers only when the concentration of FA in the medium was three times the standard concentration, which was expected to have a protective effect against MTX. These results show the importance of nutrient concentrations, exposure scenarios and timing of read-outs for cell differentiation and compound sensitivity in the ESTn. Caution should be taken when interpreting results from a single in vitro test, especially when extrapolating to effects on complex morphogenetic processes, like neural tube development.

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Methotrexate and Valproic Acid Affect Early Neurogenesis of Human Amniotic Fluid Stem Cells from Myelomeningocele

Cited by 9 publications

References 39 publications

Reduced H3K27me3 Suppresses Wnt/β-catenin Signaling by S-adenosylmethionine Deficiency in Neural Tube Development

Reduced H3K27me3 Suppresses Wnt/β-catenin Signaling by S-adenosylmethionine Deficiency in Neural Tube Development

Transcriptional Signature of Valproic Acid-Induced Neural Tube Defects in Human Spinal Cord Organoids

Culture Conditions Affect Chemical-Induced Developmental Toxicity In Vitro: The Case of Folic Acid, Methionine and Methotrexate in the Neural Embryonic Stem Cell Test

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