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1.
Heliyon ; 9(11): e21915, 2023 Nov.
Article in English | MEDLINE | ID: mdl-38034615

ABSTRACT

Nkx2.3, a transcription factor, plays important roles in various developmental processes. However, the mechanisms underlying nkx2.3's regulation of pouch and pharyngeal arch development in zebrafish remain unclear. In this study, we demonstrated that knockdown or knockout of nkx2.3 resulted in the absence of posterior ceratobranchial cartilages in zebrafish. The absence of posterior pharyngeal cartilages is a consequence of the compromised proliferation and differentiation and survival of cranial neural crest cells (CNCCs). Notably, we found that nkx2.3 was not involved in endoderm pouch formation. Additionally, our findings suggested that nkx2.3 negatively regulated Fibroblast growth factor (Fgf) signaling, as overexpression of fgf8 could mimic the phenotype observed in nkx2.3 morphants, suppressing CNCC differentiation. Moreover, inhibiting Fgf signaling restored the abnormalities in posterior cartilages induced by nkx2.3 knockdown. These findings establish the essential role of nkx2.3 in the development of posterior ceratobranchial cartilages through the inhibition of fgf8.

2.
iScience ; 26(9): 107516, 2023 Sep 15.
Article in English | MEDLINE | ID: mdl-37636049

ABSTRACT

Adducin 1 (Add1) is known as a membrane cytoskeletal protein, but its nuclear function remains unclear. In this study, we generated add1-deficient zebrafish to investigate its role in hematopoiesis. Lack of add1 impaired both primitive and definitive hematopoiesis, preventing healthy erythrocyte development. RNA sequencing revealed activation of the p53 pathway in add1-depleted erythroblast cells, leading to apoptosis at the 14-somites stage and 24 hpf. Interestingly, partial rescue of the anemic phenotype and apoptosis was observed with p53 insufficiency. Mechanistically, ADD1 was found to regulate promoter activity. These findings demonstrate that Add1 plays a crucial role in zebrafish erythropoiesis, involving the p53-mediated apoptotic pathway, expanding its regulatory role beyond cytoskeletal functions.

3.
Hum Hered ; 88(1): 29-37, 2023.
Article in English | MEDLINE | ID: mdl-36944328

ABSTRACT

INTRODUCTION: We have reported that high total homocysteine and the coexistence of inadequate thyroid hormones in maternal serum increase the risk of fetal neural tube defects (NTDs). Placental iodothyronine deiodinases (DIOs: DIO1, DIO2, and DIO3) play a role in regulating the conversions between different forms of maternal thyroid hormones. This study hypothesized that single nucleotide polymorphisms (SNPs) in placental DIOs genes could be related to NTDs. METHODS: We performed a case-control study from 2007 to 2009 that included pregnant women from Lüliang, Shanxi Province, China. Nine distinct SNPs in DIOs genes were analyzed, and placental samples were obtained from 83 pregnant women with NTD fetuses and 90 pregnant women with normal fetuses. The nine SNPs were analyzed using the Cochran-Armitage test and the Fisher's exact test. RESULTS: There were no statistically significant differences between case and control in the nine SNPs of DIOs (p > 0.05). CONCLUSIONS: The results of this study suggested that SNPs of DIO genes in the placenta among pregnant women have no statistically significant difference between the two groups, suggesting that other factors might be involved in metabolism of maternal thyroid hormone provided to fetuses, such as epigenetic modification of methylation and homocysteinylation and genomic imprinting in the placenta. Further functional studies on placenta samples are necessary.


Subject(s)
Neural Tube Defects , Placenta , Pregnancy , Humans , Female , Placenta/metabolism , Iodide Peroxidase/genetics , Iodide Peroxidase/metabolism , Case-Control Studies , Prevalence , Thyroid Hormones/metabolism , Neural Tube Defects/epidemiology , Neural Tube Defects/genetics , Neural Tube Defects/metabolism , China/epidemiology
4.
Brain ; 146(8): 3455-3469, 2023 08 01.
Article in English | MEDLINE | ID: mdl-36928982

ABSTRACT

Neural tube defects are the most severe congenital malformations that result from failure of neural tube closure during early embryonic development, and the underlying molecular mechanisms remain elusive. Retinoic acid, an active derivative of vitamin A, is critical for neural system development, and retinoic acid receptor (RAR) signalling malfunctions have been observed in human neural tube defects. However, retinoic acid-retinoic acid receptor signalling regulation and mechanisms in neural tube defects are not fully understood. The mRNA expression of RARs and retinoid X receptors in the different human neural tube defect phenotypes, including 11 pairs of anencephaly foetuses, 10 pairs of hydrocephalus foetuses and nine pairs of encephalocele foetuses, was investigated by NanoString nCounter technology. Immunoprecipitation-mass spectrometry was performed to screen the potential interacting targets of retinoic acid receptor γ. The interactions between proteins were confirmed by co-immunoprecipitation and immunofluorescence laser confocal microscopy. Luciferase and chromatin immunoprecipitation with quantitative real-time polymerase chain reaction assays were used to clarify the underlying mechanism. Moreover, a neural tube defect animal model, constructed using excess retinoic acid, was used for further analysis with established molecular biology technologies. We report that level of retinoic acid receptor γ (RARγ) mRNA was significantly upregulated in the brain tissues of human foetuses with anencephaly. To further understand the actions of retinoic acid receptor γ in neural tube defects, methylenetetrahydrofolate dehydrogenase 1 was identified as a specific retinoic acid receptor γ target from IP-MS screening. Additionally, methylenetetrahydrofolate dehydrogenase 1 negatively regulated retinoic acid receptor γ transcription factor activity. Furthermore, low expression of methylenetetrahydrofolate dehydrogenase 1 and activation of retinoic acid receptor signalling were further determined in human anencephaly and a retinoic acid-induced neural tube defect mouse model. This study reveals that methylenetetrahydrofolate dehydrogenase 1, the rate-determining enzyme in the one-carbon cycle, might be a specific regulator of retinoic acid receptors; these findings provide new insights into the functional linkage between nuclear folate metabolism and retinoic acid receptor signalling in neural tube defect pathology.


Subject(s)
Anencephaly , Neural Tube Defects , Mice , Pregnancy , Animals , Female , Humans , Methylenetetrahydrofolate Dehydrogenase (NADP)/adverse effects , Receptors, Retinoic Acid/genetics , Receptors, Retinoic Acid/metabolism , Tretinoin/adverse effects , Neural Tube Defects/chemically induced , Neural Tube Defects/genetics , Neural Tube Defects/metabolism , RNA, Messenger , Minor Histocompatibility Antigens
5.
J Trace Elem Med Biol ; 68: 126814, 2021 Dec.
Article in English | MEDLINE | ID: mdl-34243025

ABSTRACT

BACKGROUND: This study tested the hypothesis that abnormal maternal metabolism of both homocysteine and thyroid hormone network in pregnant women is associated with neural tube defects (NTDs) in a part of China with high NTD prevalence. METHODS: A case-control study was performed between 2007 and 2009 in Lüliang Mountains, Shanxi Province. This study included 83 pregnant women who had fetuses with NTDs (cases) and 90 pregnant women with normal fetuses (controls). In addition, a cell model to illustrate the epidemiological findings was established. RESULTS: Fetuses of mother who had both high total homocysteine (tHcy) and inadequate free thyroxine were 3 times more at risk of developing NTDs (adjusted odds ratio = 3.5; 95 % confidence interval = 1.2-10.4; cases vs. controls) using multivariate logistic regression models. Furthermore, biological interaction between metabolisms of Hcy and thyroid hormones was demonstrated in vitro. In homocysteine thiolactone of a metabolite of Hcy-treated mouse embryonic neural stem NE4C cells, genes (Bmp7, Ctnnb1, Notch 1, Gli2, and Rxra) related to both neural tube closure and thyroid hormone network were shown to be regulated by H3K79 homocysteinylation, which increased their expression levels. CONCLUSIONS: The effect of maternal serum high tHcy on risk of developing NTDs is depended on maternal serum level of thyroxine. Meanwhile, a higher level of tHcy might also affect both maternal metabolism of thyroid hormone and neural tube closure in embryogenesis through homocysteinylation of histones.


Subject(s)
Neural Tube Defects , Pregnant Women , Animals , Case-Control Studies , Female , Folic Acid , Homocysteine , Humans , Mice , Pregnancy , Thyroid Hormones , Thyroxine
6.
Cell Death Dis ; 12(3): 234, 2021 03 04.
Article in English | MEDLINE | ID: mdl-33664222

ABSTRACT

Wnt signaling plays a major role in early neural development. An aberrant activation in Wnt/ß-catenin pathway causes defective anteroposterior patterning, which results in neural tube closure defects (NTDs). Changes in folate metabolism may participate in early embryo fate determination. We have identified that folate deficiency activated Wnt/ß-catenin pathway by upregulating a chorion-specific transcription factor Gcm1. Specifically, folate deficiency promoted formation of the Gcm1/ß-catenin/T-cell factor (TCF4) complex formation to regulate the Wnt targeted gene transactivation through Wnt-responsive elements. Moreover, the transcription factor Nanog upregulated Gcm1 transcription in mESCs under folate deficiency. Lastly, in NTDs mouse models and low-folate NTDs human brain samples, Gcm1 and Wnt/ß-catenin targeted genes related to neural tube closure are specifically overexpressed. These results indicated that low-folate level promoted Wnt/ß-catenin signaling via activating Gcm1, and thus leaded into aberrant vertebrate neural development.


Subject(s)
DNA-Binding Proteins/metabolism , Folic Acid Deficiency/metabolism , Neural Tube Defects/metabolism , Neural Tube/metabolism , Transcription Factors/metabolism , Wnt Signaling Pathway , Animals , DNA-Binding Proteins/genetics , Disease Models, Animal , Female , Folic Acid Deficiency/complications , Folic Acid Deficiency/genetics , Gene Expression Regulation, Developmental , HEK293 Cells , Humans , Mice , Mice, Inbred C57BL , Nanog Homeobox Protein/genetics , Nanog Homeobox Protein/metabolism , Neural Tube/abnormalities , Neural Tube Defects/etiology , Neural Tube Defects/genetics , Pregnancy , Transcription Factor 4/genetics , Transcription Factor 4/metabolism , Transcription Factors/genetics
8.
Opt Express ; 28(21): 31057-31075, 2020 Oct 12.
Article in English | MEDLINE | ID: mdl-33115089

ABSTRACT

Traditional cameras are limited by sensors and cannot directly capture single-shot high dynamic range (HDR) images. We propose an improved single-shot HDR image reconstruction method that uses a single-exposure filtered low dynamic range (FLDR) image. First, by adding an optical filter in front of the camera lens, a FLDR image with different RGB channel exposure states and luminance ranges can be captured in a single-shot, unlike the traditional LDR image. Second, a deep inverse tone mapping network (DITMnet) with multibranch features extraction and multioutput images synthesis is designed to reconstruct an HDR image from a single FLDR image. Experimentally, under different exposure states and color spaces, our method outperforms similar algorithms.

9.
Epigenomics ; 12(1): 5-18, 2020 01.
Article in English | MEDLINE | ID: mdl-31769301

ABSTRACT

Aim: To know the cause of sequence variants in neural tube defect (NTD). Materials & methods: We sequenced genes implicated in neural tube closure (NTC) in a Chinese cohort and elucidated the molecular mechanism-driving mutations. Results: In NTD cases, an increase in specific variants was identified, potentially deleterious rare variants harbored in H3K36me3 occupancy regions that recruits mismatch repair (MMR) machinery. Lower folate concentrations in local brain tissues were also observed. In neuroectoderm cells, folic acid insufficiency attenuated association of Msh6 to H3K36me3, and reduced bindings to NTC genes. Rare variants in human NTDs were featured by MMR deficiency and more severe microsatellite instability. Conclusion: Our work suggests a mechanistic link between folate insufficiency and MMR deficiency that correlates with an increase of rare variants in NTC genes.


Subject(s)
DNA Mismatch Repair/genetics , Disease Susceptibility , Folic Acid Deficiency/complications , Neural Tube Defects/etiology , Neural Tube Defects/metabolism , Folic Acid/metabolism , Folic Acid Deficiency/metabolism , Genetic Variation , Histones/metabolism , Humans
10.
FASEB J ; 33(4): 4688-4702, 2019 04.
Article in English | MEDLINE | ID: mdl-30592646

ABSTRACT

Folate deficiency in early development leads to disturbance in multiple processes, including neurogenesis during which fibroblast growth factor (FGF) pathway is one of the crucial pathways. Whether folic acid (FA) directly affects FGF pathways to influence neurodevelopment and the possible mechanism remains unclear. In this study, we presented evidence that in human FA-insufficient encephalocele, the FGF pathway was interfered. Furthermore, in Brachyury knockout mice devoid of such T-box transcription factors regulating embryonic neuromesodermal bipotency and a key component of FGF pathway, change in expression of Brachyury downstream targets, activator Fgf8 and suppressor dual specificity phosphatase 6 was detected, along with the reduction in expression of other key FGF pathway genes. By using a FA-deficient cell model, we further demonstrated that decrease in Brachyury expression was through alteration in hypermethylation at the Brachyury promoter region under FA deficiency conditions, and suppression of Brachyury promoted the inactivation of the FGF pathway. Correspondingly, FA supplementation partially reverses the effects seen in FA-deficient embryoid bodies. Lastly, in mice with maternal folate-deficient diets, aberrant FGF pathway activity was found in fetal brain dysplasia. Taken together, our findings highlight the effect of FA on FGF pathways during neurogenesis, and the mechanism may be due to the low expression of Brachyury gene via hypermethylation under FA-insufficient conditions.-Chang, S., Lu, X., Wang, S., Wang, Z., Huo, J., Huang, J., Shangguan, S., Li, S., Zou, J., Bao, Y., Guo, J., Wang, F., Niu, B., Zhang, T., Qiu, Z., Wu, J., Wang, L. The effect of folic acid deficiency on FGF pathway via Brachyury regulation in neural tube defects.


Subject(s)
Fetal Proteins/metabolism , Fibroblast Growth Factors/metabolism , Folic Acid Deficiency/metabolism , Folic Acid/therapeutic use , Neural Tube Defects/drug therapy , Neural Tube Defects/metabolism , T-Box Domain Proteins/metabolism , Animals , Apoptosis/drug effects , Blotting, Western , Cell Proliferation/drug effects , Cells, Cultured , Chromatin Immunoprecipitation , Encephalocele/metabolism , Female , Folic Acid Deficiency/physiopathology , Gene Expression Regulation, Developmental/drug effects , Humans , Immunohistochemistry , Mice , Mice, Inbred C57BL , Mice, Knockout , Promoter Regions, Genetic , Signal Transduction/drug effects , Sulfites/pharmacology
11.
Wei Sheng Yan Jiu ; 47(6): 883-889, 2018 Nov.
Article in Chinese | MEDLINE | ID: mdl-30593316

ABSTRACT

OBJECTIVE: To analysis the associations between the polymorphisms of TMPRSS6 and the levels of serum ferritin( SF) and soluble transferrin receptor( s TfR) in pregnant women. METHODS: A total of 807 pregnant women were recruited by using cluster random sampling method from Lüliang in Shanxi Province in 2014. Roche Tinaquant immunoturbidimetric assay was used to measure the level of SF and s TfR, then iron deficiency( ID) was identified by the criteria of SF < 25 ng/m L and s TfR > 4. 4 mg/L, respectively. Sequenom MassArray was used to genotype the 7 targeted single-nucleotide polymorphisms( SNPs)( rs11704654, rs1421312, rs2111833, rs2235321, rs2543519, rs4820268 and rs855791) of the subjects. The t test and ANOVA analysis were used to test the different levels of SF and s TfR among SNPs, and chi-square test and Logistic regression were conducted to detect the associations between genotypes of each locus and ID. RESULTS: The differences of the levels of Ln SF between genotypes in rs2111833 were significant( F = 3. 57, P = 0. 0287), and the Ln SF level of T allele carrier group was lower than CC group( t = 2. 03, P = 0. 0429). The Ln SF level of A allele carrier group was lower than GG carrier group in rs855791( t = 1. 97, P = 0. 0490). For rs11704654, the ratio of SF < 25 ng/m L of T allele carriers was higher than CC carriers( χ~2= 4. 5456, P = 0. 0330). For rs211183, the ratio of SF < 25 ng/m L of T allele carriers was higher than CC carriers( χ~2= 4. 6431, P = 0. 0312). For rs855791, the ratio of SF < 25 ng/m L of GG carriers was lower than A allele carriers( χ~2= 5. 0134, P = 0. 0263). rs11704654( T) and rs855791( A) were still shown the association with SF < 25 ng/m L status in logistic analysis adjusted by age and gestational weeks. The Ln s TfR level of T allele carrier group in rs11704654 was higher than CC carrier group( t =-2. 012, P = 0. 024), and the Ln s TfR level of G allele carrier group in rs2543519 was higher than AA carrier group( t =-1. 954, P = 0. 011). CONCLUSION: The associations between polymorphisms of TMPRSS6 and the levels of SF and s TfR are observed in pregnant women.


Subject(s)
Ferritins , Membrane Proteins , Polymorphism, Single Nucleotide , Receptors, Transferrin , Serine Endopeptidases , Alleles , Female , Ferritins/blood , Humans , Membrane Proteins/genetics , Pregnancy , Receptors, Transferrin/blood , Serine Endopeptidases/genetics
12.
BMC Med Genomics ; 11(1): 38, 2018 04 04.
Article in English | MEDLINE | ID: mdl-29618362

ABSTRACT

BACKGROUND: Mouse homozygous mutants in Wnt/planar cell polarity (PCP) pathway genes have been shown to cause neural tube defects (NTDs) through the disruption of normal morphogenetic processes critical to neural tube closure (NTC). Knockout mice that are heterozygotes of single PCP genes likely fail to produce NTD phenotypes, yet damaging variants detected in human NTDs are almost always heterozygous, suggesting that other deleterious interacting variants are likely to be present. Nonetheless, the Wnt/PCP pathway remains a genetic hotspot. Addressing these issues is essential for understanding the genetic etiology of human NTDs. METHODS: We performed targeted next-generation sequencing (NGS) on 30 NTD-predisposing Wnt/PCP pathway genes in 184 Chinese NTD cases. We subsequently replicated our findings for the CELSR1 gene in an independent cohort of 292 Caucasian NTD samples from the USA. Functional validations were confirmed using in vitro assays. RESULTS: CELSR1, CELSR2 and CELSR3 genes were significantly clustered with rare driver coding mutations (q-value< 0.05) demonstrated by OncodriveCLUST. During the validation stage, the number of rare loss of function (LoF) variants in CELSR1 was significantly enriched in NTDs compared with the LoF counts in the ExAC database (p < 0.001). Functional studies indicated compound heterozygote variants of CELSR2 p.Thr2026Met and DVL3 p.Asp403Asn result in down regulation of PCP signals. CONCLUSIONS: These data indicate rare damaging variants of the CELSR genes, identified in ~ 14% of NTD cases, are expected to be driver genes in the Wnt/PCP pathway. Compound damaging variants of CELSR genes and other Wnt/PCP genes, which were observed in 3.3% of the studied NTD cohort, are also expected to amplify these effects at the pathway level.


Subject(s)
Cell Polarity/genetics , Neural Tube Defects/genetics , Neural Tube Defects/pathology , Wnt Proteins/genetics , Cadherins/genetics , Child , Genetic Predisposition to Disease , HEK293 Cells , HeLa Cells , Humans , Mutation
13.
CNS Neurosci Ther ; 24(5): 394-403, 2018 05.
Article in English | MEDLINE | ID: mdl-29365368

ABSTRACT

AIM: Neural tube defects (NTDs) are birth defects of the nervous system and are the second most frequent cause of birth defects worldwide. The etiology of NTDs is complicated and involves both genetic and environmental factors. CASP9 is an initiator caspase in the intrinsic apoptosis pathway, which in Casp9-/- mice has been shown to result in NTDs because of decreased apoptosis. The aim of this study was to evaluate the potential genetic contribution of the CASP9 gene in human NTDs. METHODS: High-throughput sequencing was performed to screen genetic variants of CASP9 genes in 355 NTD cases and 225 matched controls. Apoptosis-relevant assays were performed on transiently transfected E9 neuroepithelial cells or human embryonic kidney 293T cells, to determine the functional characteristics of NTD-specific rare variants under complete or low folic acid (FA) status. RESULTS: We found significant expression of CASP9 rare variants in NTDs and identified 4 NTD-specific missense variants. Functional assays demonstrated that a p.Y251C variant attenuates apoptosis by reducing CASP9 protein expression and decreasing activity of the intrinsic apoptosis pathway. From this, we conclude that this variant may represent a loss-of-function mutation. A 4-time recurrent p.R191G variant did not affect intrinsic apoptosis in complete medium, while it completely inhibited apoptosis induced by low FA medium. CONCLUSION: Our findings identify a genetic link for apoptosis in human NTDs and highlight the effect of gene-environment interactions in a complex disease.


Subject(s)
Caspase 9/genetics , Caspase 9/metabolism , Mutation , Neural Tube Defects/genetics , Neural Tube Defects/metabolism , Apoptosis/physiology , Asian People/genetics , Cell Line , China , Cohort Studies , Female , Folic Acid/metabolism , Folic Acid Deficiency/metabolism , Gene Expression , Gene-Environment Interaction , Genetic Testing , Humans , Male
14.
Cell Mol Neurobiol ; 38(3): 605-614, 2018 Apr.
Article in English | MEDLINE | ID: mdl-28623428

ABSTRACT

Although more than 200 genes are known to be related to neural tube defects (NTDs), the exact molecular basis is still unclear. Evaluating the contribution of copy number variation (CNV) might be a priority because CNV involves changes in the copy number of large segments of DNA, leading to phenotypic traits and disease susceptibility. Recent studies have documented that the polarity protein partitioning defective 3 homolog (Pard3) plays an essential role in the process of neural tube closure. The aim of this study was to assess the role of PARD3 CNVs in the etiology of human NTDs. Relative quantitative PCR and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry were used to quantitative measurement of CNVs in 25 PARD3 exons in 202 NTD cases and 231 controls from a region of China with a high prevalence of NTDs. The results showed that microduplications ranging from 3 to 4 were evident in coding Exon 21 and Exon 25 in both case and control groups. A novel heterozygous microdeletion spanning 444 bp of Exon 14 was identified in two cases of anencephaly and is absent from all controls analyzed. Expression analyses indicated that this heterozygotic microdeletion showed no tissue specificity and led to defective expression of PARD3. Our study provides further evidence implicating PARD3 in the etiology of NTDs.


Subject(s)
Cell Cycle Proteins/genetics , DNA Copy Number Variations/genetics , Genetic Predisposition to Disease/genetics , Membrane Proteins/genetics , Neural Tube Defects/genetics , Adaptor Proteins, Signal Transducing , Adult , Cell Cycle Proteins/metabolism , Female , Humans , Male , Membrane Proteins/metabolism , Middle Aged , Real-Time Polymerase Chain Reaction/methods
15.
Mol Neurobiol ; 54(8): 6304-6316, 2017 Oct.
Article in English | MEDLINE | ID: mdl-27714636

ABSTRACT

Dishevelled (DVL/Dvl) genes play roles in canonical and noncanonical Wnt signaling, both of which are essential in neural tube closing and are involved in balancing neural progenitor growth and differentiation, or neuroepithelial cell polarity, respectively. In mouse Dvl haploinsufficiency leads to neural tube defects (NTDs), which represent the second most common birth defects. However, DVL genes' genetic contributions in human NTDs are modest. We sought to explore the molecular impact on such genes in human NTDs in a Han Chinese cohort. In 47 cases with NTDs and 61 matched controls, in brain tissues, the DVL1/2 mRNA levels were correlated with the levels of a serine/threonine protein kinase MARK2, and in 20 cases with lumbosacral spina bifida, the mRNA levels of DVL1 and MARK2 were significantly decreased; by contrast, only an intronic rare variant was found. Moreover, in an extended population, we found merely three novel rare missense variants in 1 % of individuals with NTDs. In cell-based assays, Mark2 depletion indeed reduces Dvl gene expression and interrupts neural stem cell (NSCs) growth and differentiation, which are likely to be mediated through a decrease in class IIa HDAC phosphorylation and reduced H3K4ac and H3K27ac occupancies at the Dvl1/2 promoters. Finally, the detections of folate concentration in human brain tissue and NSCs and MEF cells indicates that folate deficiency contributes to the observed decreases in Mark2 and Dvl1 expression. Our present study raises a potential common pathogenicity mechanism in human lumbosacral spina bifida about DVL genes rather than their genetic pathogenic role.


Subject(s)
Dishevelled Proteins/genetics , Histones/metabolism , Protein Serine-Threonine Kinases/genetics , Spinal Dysraphism/genetics , Transcription, Genetic , Acetylation , Cell Proliferation/physiology , Child , Child, Preschool , Dishevelled Proteins/metabolism , Female , Humans , Infant , Infant, Newborn , Male , Neural Stem Cells/metabolism , Neurogenesis/physiology , Phosphorylation , Promoter Regions, Genetic , Protein Serine-Threonine Kinases/metabolism , Spinal Dysraphism/metabolism
16.
Hum Mutat ; 38(4): 378-389, 2017 04.
Article in English | MEDLINE | ID: mdl-27925688

ABSTRACT

Increasing evidence that mutation of planar cell polarity (PCP) genes contributes to human cranial neural tube defect (NTD) susceptibility prompted us to hypothesize that rare variants of genes in the core apical-basal polarity (ABP) pathway are risk factors for cranial NTDs. In this study, we screened for rare genomic variation of PARD3 in 138 cranial NTD cases and 274 controls. Overall, the rare deleterious variants of PARD3 were significantly associated with increased risk for cranial NTDs (11/138 vs.7/274, P < 0.05, OR = 3.3). These NTD-specific variants were significantly enriched in the aPKC-binding region (6/138 vs. 0/274, P < 0.01). The East Asian cohort in the ExAC database and another Chinese normal cohort further supported this association. Over-expression analysis in HEK293T and MDCK cells confirmed abnormal aPKC binding or interaction for two PARD3 variants (p.P913Q and p.D783G), resulting in defective tight junction formation via disrupted aPKC binding. Functional analysis in human neural progenitor cells and chick embryos revealed that PARD3 knockdown gave rise to abnormal cell polarity and compromised the polarization process of neuroepithelial tissue. Our studies suggest that rare deleterious variants of PARD3 in the aPKC-binding region contribute to human cranial NTDs, possibly by disrupting apical tight junction formation and subsequent polarization process of the neuroepithelium.


Subject(s)
Cell Cycle Proteins/genetics , Membrane Proteins/genetics , Mutation , Neural Tube Defects/genetics , Protein Kinase C/metabolism , Tight Junctions/metabolism , Adaptor Proteins, Signal Transducing , Animals , Asian People/genetics , Body Patterning/genetics , Cell Cycle Proteins/metabolism , Chick Embryo , China , Cohort Studies , Dogs , HEK293 Cells , Humans , Madin Darby Canine Kidney Cells , Membrane Proteins/metabolism , Neural Tube Defects/ethnology , Neural Tube Defects/metabolism , Protein Binding , RNA Interference , Tight Junctions/pathology
17.
G3 (Bethesda) ; 6(10): 3307-3316, 2016 10 13.
Article in English | MEDLINE | ID: mdl-27543293

ABSTRACT

Neural tube defects (NTDs) are severe malformations of the central nervous system caused by complex genetic and environmental factors. Among genes involved in NTD, cilia-related genes have been well defined and found to be essential for the completion of neural tube closure (NTC). We have carried out next-generation sequencing on target genes in 373 NTDs and 222 healthy controls, and discovered eight disease-specific rare mutations in cilia-related gene DNAAF1 DNAAF1 plays a central role in cytoplasmic preassembly of distinct dynein-arm complexes, and is expressed in some key tissues involved in neural system development, such as neural tube, floor plate, embryonic node, and brain ependyma epithelial cells in zebrafish and mouse. Therefore, we evaluated the expression and functions of mutations in DNAAF1 in transfected cells to analyze the potential correlation of these mutants to NTDs in humans. One rare frameshift mutation (p.Gln341Argfs*10) resulted in significantly diminished DNAAF1 protein expression, compared to the wild type. Another mutation, p.Lys231Gln, disrupted cytoplasmic preassembly of the dynein-arm complexes in cellular assay. Furthermore, results from NanoString assay on mRNA from NTD samples indicated that DNAAF1 mutants altered the expression level of NTC-related genes. Altogether, these findings suggest that the rare mutations in DNAAF1 may contribute to the susceptibility for NTDs in humans.


Subject(s)
Cilia/genetics , Genetic Association Studies , Mutation , Neural Tube Defects/genetics , Adolescent , Adult , Alleles , Amino Acid Sequence , Amino Acid Substitution , Case-Control Studies , Cell Line , DNA Mutational Analysis , Female , Gene Expression , Genetic Variation , Genotype , Humans , Male , Neural Tube Defects/diagnosis , Neurogenesis/genetics , Phenotype , Young Adult
18.
Birth Defects Res A Clin Mol Teratol ; 106(4): 232-9, 2016 Apr.
Article in English | MEDLINE | ID: mdl-27001897

ABSTRACT

BACKGROUND: The polymorphism of genes involved in folate-mediated one-carbon metabolism may be a risk factor for neural tube defects (NTDs). In the present study, we aimed to investigate the single nucleotide polymorphisms (SNPs) of the genes BHMT, CUBN, FTCD, GAMT, GART, SARDH, SHMT1, and MUT, and their effect on NTDs in the Chinese Han population. METHODS: A total of 270 NTDs cases and 192 controls were enrolled in this study. The SNPs were analyzed with the next-generation sequencing method. The folate levels of brain tissues from 113 available NTDs cases and 123 available controls were measured. RESULTS: Next-generation sequencing identified 818 single nucleotide variants, including 214 SNPs used for further analysis. Statistical analysis showed that two independent SNP loci, rs2797840 and rs2073817 in SARDH, may be associated with the susceptibility of NTDs. Specifically, the minor allele G of rs2797840 was significantly associated with NTDs risk in spina bifida subgroup (p value = 0.0348). For subjects whose folate content was measured, the protective allele G of rs2797840 was significantly associated with increased folate content of brain. rs2797840 is within several ENCODE regulatory regions, indicating this SNPs may influence expression of SARDH. CONCLUSION: The SNPs rs2797840 and rs2073817 in SARDH may serve as an indicator for the occurrence of NTDs in the Chinese Han population, and rs2797840 may also be an indicator for folate content of brain.


Subject(s)
Folic Acid/genetics , Neural Tube Defects/genetics , Polymorphism, Single Nucleotide , Sarcosine Dehydrogenase/genetics , China , Female , Folic Acid/metabolism , High-Throughput Nucleotide Sequencing , Humans , Male , Neural Tube Defects/metabolism , Sarcosine Dehydrogenase/metabolism
19.
Neurol Sci ; 37(7): 1049-54, 2016 Jul.
Article in English | MEDLINE | ID: mdl-26993122

ABSTRACT

We investigated single-nucleotide polymorphisms (SNPs) in the aldehyde dehydrogenase family1 L1 gene (ALDH1L1) and their association with neural tube defects (NTDs) in the Chinese population. A total of 271 NTDs cases and 192 healthy controls were used in this study. A total of 112 selected SNPs in the ALDH1L1 gene were analyzed using the next-generation sequencing method. Statistical analysis was carried out to investigate the correlation between SNPs and patient susceptibility to NTDs. Statistical analysis revealed a significant correlation between the SNP sites rs4646733, rs2305225, and rs2276731 in the ALDH1L1 gene and NTDs. The TT genotype and T allele of rs4646733 in ALDH1L1 were associated with a significantly increased incidence of NTDs [odds ratio (OR) = 2.16, 95 % confidence interval (CI) 1.199-3.896 for genotype; and OR = 1.46, 95 % CI 1.092-1.971 for allele]. The AA genotype and A allele of rs2305225 in ALDH1L1 were associated with a significantly increased incidence of NTDs (OR = 2.03, 95 % CI 1.202-3.646 for genotype, and OR = 1.44, 95 % CI 1.096-1.905 for allele). The CT genotype and C allele of rs2276731 in ALDH1L1 significantly were associated with an increased incidence of NTDs (OR = 1.67, 95 % CI 1.129-2.491 with genotype, and OR = 1.32, 95 % CI 0.956-1.816 with allele).The polymorphic loci rs4646733, rs2305225, and rs2276731 in the ALDH1L1 gene maybe potential risk factors for NTDs in the Chinese population.


Subject(s)
Aldehyde Dehydrogenase/genetics , Genetic Predisposition to Disease/ethnology , Genetic Predisposition to Disease/genetics , Neural Tube Defects/genetics , Polymorphism, Single Nucleotide/genetics , Alleles , Asian People/ethnology , Asian People/genetics , Female , Genetic Association Studies , Genotype , Humans , Male , Oxidoreductases Acting on CH-NH Group Donors , Risk Factors
20.
Mol Neurobiol ; 53(8): 5413-24, 2016 10.
Article in English | MEDLINE | ID: mdl-26446020

ABSTRACT

GLI2 is a key mediator of the sonic hedgehog (Shh) signaling pathway and plays an important role in neural tube development during vertebrate embryogenesis; however, the role of gli2 in human folate-related neural tube defects remains unclear. In this study, we compared methylation status and polymorphisms of gli2 between spina bifida patients and a control group to explore the underlying mechanisms related to folate deficiency in spina bifida. No single nucleotide polymorphism was found to be significantly different between the two groups, although gli2 methylation levels were significantly increased in spina bifida samples, accompanied by aberrant GLI2 expression. Moreover, a prominent negative correlation was found between the folate level in brain tissue and the gli2 methylation status (r = -0.41, P = 0.014), and gli2 hypermethylation increased the risk of spina bifida with an odds ratio of 12.45 (95 % confidence interval: 2.71-57.22, P = 0.001). In addition, we established a cell model to illustrate the effect of gli2 expression and the accessibility of chromatin affected by methylation. High gli2 and gli1 mRNA expression was detected in 5-Aza-treated cells, while gli2 hypermethylation resulted in chromatin inaccessibility and a reduced association with nuclear proteins containing transcriptional factors. More meaningful to the pathway, the effect gene of the Shh pathway, gli1, was found to have a reduced level of expression along with a decreased expression of gli2 in our cell model. Aberrant high methylation resulted in the low expression of gli2 in spina bifida, which was affected by the change in chromatin status and the capacity of transcription factor binding.


Subject(s)
DNA Methylation/genetics , Gene Expression Regulation, Developmental , Hedgehog Proteins/metabolism , Nuclear Proteins/genetics , Promoter Regions, Genetic , Signal Transduction , Spinal Dysraphism/genetics , Zinc Finger Protein Gli2/genetics , Adult , Brain/metabolism , Brain/pathology , Case-Control Studies , Chromatin/metabolism , Female , Folic Acid/metabolism , Humans , Male , Nuclear Proteins/metabolism , Polymorphism, Single Nucleotide/genetics , RNA, Messenger/genetics , RNA, Messenger/metabolism , Signal Transduction/genetics , Transcription, Genetic , Zinc Finger Protein GLI1/genetics , Zinc Finger Protein GLI1/metabolism , Zinc Finger Protein Gli2/metabolism
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