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List of rare coding PCP variants identified in neural tube defects (NTDs) and predicted to be damaging and/or affect protein function in NTDs.
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Neural tube defects (NTDs), including spina bifida and anencephaly, represent the most severe and common malformations of the central nervous system affecting 0.7–3 per 1000 live births. They result from the failure of neural tube closure during the first few weeks of pregnancy. They have a complex etiology that implicate a large number of genetic...
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... majority of these studies included genetic as well as functional data in cell cultures and zebrafish models. Table 1 summarizes the major positive findings from these studies. It lists rare PCP variants that were associated with NTDs and that were predicted to be pathogenic and/or functionally validated. ...Context 2
... mice, single mutants of PCP genes suffer mainly from the severe CRS and rarely exhibit open spina bifida [38,41,56,57,[59][60][61]. CRS is an extremely rare type of NTDs in humans hindering the availability of large cohorts for genetic studies. Assuming no overlap in the cohorts included in Table 1, 8 PCP genes (VANGL1-2, CELSR1-3, DVL3, SCRIB1 and PTK7) have been re-sequenced in a couple of hundred of CRS cases and no obvious loss of function (LoF) variant was identified in these genes in the majority of CRS cases analyzed. Interestingly, the double mutants of PCP genes in mice exhibit a wide range of open NTDs including anencephaly and open spina bifida, providing a strong rationale for a comprehensive analysis of this pathway in other types of open NTDs in humans [38,62]. ...Context 3
... large number of studies have been conducted on the role of PCP signaling in open NTDs with a focus on the most common type that is MMC. As shown in Table 1, potentially pathogenic rare variants were identified in each of the 14 PCP genes in a small fraction of open NTD patients. As in the case for CRS, the majority of these variants were missense with no obvious LoF effect. ...Context 4
... in the case for CRS, the majority of these variants were missense with no obvious LoF effect. Further shown in Table 1, a few PCP studies have also included closed NTDs and obtained similar types of genetic variants, suggesting a shared role for this pathway in the pathogenesis of open and closed forms of NTDs. ...Similar publications
Citations
... In vertebrates and mammals, the Wnt/PCP signaling pathway controls various morphogenetic processes in most tissues and provides positional cues to cells for directed movements, particularly during gastrulation and neurulation 30 . Mutations in Vangl2 are associated with severe disorders of the nervous system called neural tube defects (NTDs), such as spina bifida and anencephaly 31 . During these embryonic processes, Vangl2 is thought to regulate Wnt/PCP signaling in a dosage-sensitive manner, including at the local level 32 . ...
... We show that the early deletion of vangl2 in the Emx1-Vangl2 mice has long-term consequences on the function of MfB/TE synapses, leading to disruption of DG-CA3 circuit function and a variety of impairments in hippocampal-dependent tasks. We do not exclude that other parts of the hippocampal circuitry -or beyond the hippocampal circuitry -are affected and contribute to the cognitive deficits we report 31,55 . Nevertheless, this work and our previous study 13 demonstrate the importance of Vangl2 in the architecture and function of DG-CA3 connectivity involved in declarative memory tasks. ...
In the hippocampus, dentate gyrus granule cells connect to CA3 pyramidal cells via their axons, the mossy fibers (Mf). The synaptic terminals of Mfs (Mf boutons, MfBs) form large and complex synapses with thorny excrescences (TE) on the proximal dendrites CA3 pyramidal cells (PCs). MfB/TE synapses have distinctive “detonator” properties due to low intitial release probability and large presynaptic facilitation. The molecular mechanisms shaping the morpho-functional properties of MfB/TE synapses are still poorly understood, though alterations in their morphology are associated with Down syndrome, intellectual disabilities, and Alzheimer’s disease. Here, we identify the core PCP gene Vangl2 as essential to the morphogenesis and function of MfB/TE synapses. Vangl2 colocalises with the presynaptic heparan sulfate proteoglycan glypican 4 (GPC4) to stabilise the postsynaptic orphan receptor GPR158. Embryonic loss of Vangl2 disrupts the morphology of MfBs and TEs, impairs ultrastructural and molecular organisation, resulting in defective synaptic transmission and plasticity. In adult, the early loss of Vangl2 results in a number of hippocampus-dependent memory deficits including characteristic flexibility of declarative memory, organisation and retention of working/ everyday-like memory. These deficits also lead to abnormal generalisation of memories to salient cues and diminished ability to form detailed contextual memories. Together, these results establish Vangl2 as a key regulator of DG-CA3 connectivity and functions.
Highlights
Vangl2 is a key regulator of MfB/TE synapses morphogenesis and plasticity in CA3
Vangl2-mediated GPC4-GPR158 interaction maintains MfB-TE pre-synaptic morphology and function
Vangl2 deletion affects declarative memory in adult mice
Vangl2 function is necessary for contextual learning and its loss leads toa maladaptive fear memory for salient cues
... Розрізняють два різних механізми -первинну та вторинну нейруляцію. У людини первинна нейруляція відбувається вздовж більшої частини рострокаудальної осі ембріона на 3-4 тижнях вагітності; вторинна нейруляція відбувається на 5 тижні вагітності, каудально, тільки в нижній крижовій і куприковій ділянках [7,8]. ...
... У ссавців нервова пластинка представлена псевдобагатошаровим стовпчастим епітелієм [8,19]. Зовні нервова пластинка спочатку схожа на лопатоподібний лист, медіолатеральна частина якого є ширшою за передньо-задню частину, потім вона починає потовщуватися, звужуватися медіолатерально й витягуватися рострокаудально [7]. Цей морфогенетичний процес визначають як конвергентне (клітинні маси конвергують у напрямі дорзальної серединної лінії) розширення (витягування / розширення у передньо-задньому напрямі) [7,21]. ...
... Зовні нервова пластинка спочатку схожа на лопатоподібний лист, медіолатеральна частина якого є ширшою за передньо-задню частину, потім вона починає потовщуватися, звужуватися медіолатерально й витягуватися рострокаудально [7]. Цей морфогенетичний процес визначають як конвергентне (клітинні маси конвергують у напрямі дорзальної серединної лінії) розширення (витягування / розширення у передньо-задньому напрямі) [7,21]. ...
Neurulation occurs by two different mechanisms, called primary and secondary neurulation. In humans, primary neurulation occurs along most of the rostrocaudal axis of the embryo, while secondary neurulation occurs caudally, only in the lower sacral and coccygeal regions. Primary neurulation is responsible for a change in the neural plate shape, the lateral edges of which rise and then converge at the dorsal midline to merge into a tube. Initially, the neural tube, formed as a result of primary neurulation, is open at both ends through the so-called rostral and caudal neuropores. These neuropores connect the inner part of the neural tube with the environment (amniotic cavity) and later (by the end of primary neurulation) are closed. During primary neurulation, the brain and spinal cord are formed up to the upper sacral region (up to the level of junction between S1 and S2 vertebral bodies), however, the most caudal part of this anatomical region (sacral-coccygeal division of the spinal cord, conus medullaris and filum terminale) is formed at secondary neurulation. In humans, secondary neurulation occurs due to elongation and cavitation of the caudal cell mass into the medulla, which then transforms into a secondary neural tube. Thus, the main differences between primary and secondary neurulation are that the neural plate folds and invaginates into the body of the embryo and separates from the surface ectoderm, forming an underlying hollow tube in primary neurulation. Mesenchymal cell сlusters form a dense cord that undergoes mesenchymal-epithelial transition and forms cavities and an empty tube during secondary neurulation to form the terminal part of the spinal cord. Conclusions. Understanding the detailed molecular and genetic mechanisms of each stage of neurulation is relevant due to widespread congenital neural tube defects, and only perfect knowledge on each aspect of neurulation and all possible factors of potential influence on it will help to develop modern options for influencing some of them, and probably, cause a decrease in neural tube congenital defects.
... [1][2][3] At the tissue level, PCP facilitates proper embryonic development during morphogenesis and organogenesis, defects of which can lead to improper neural tube closure and congenital heart disorders. [4][5][6][7] In response to global sensory cues, PCP is established by the asymmetric localization of two transmembrane complexes to opposite sides on the apical region of the cell: cadherin epithelial growth factor (EGF) laminin-G (LAG or LamG) seven pass G-type receptor (CELSR1) + Frizzled (Fzd3/6) and CELSR1 + Vang like (Vangl1/2). [8][9][10][11][12] Initially, the complexes are randomly distributed throughout the apical region. ...
Planar cell polarity (PCP), essential to multicellular developmental processes, arises when cells polarize and
align across tissues. Central to PCP is CELSR1, an atypical cadherin featuring a long ectodomain with nine
extracellular cadherin (EC) repeats, a membrane adjacent domain (MAD10), and several characteristic adhesion
GPCR domains. Cell-based aggregation assays have demonstrated CELSR1’s homophilic adhesive
nature, but mechanistic details are missing. Here, we investigate the possible adhesive properties and structures
of CELSR1 EC repeats. Our bead aggregation assays do not support strong adhesion by EC repeats
alone. Consistently, EC1-4 only dimerizes at high concentration in solution. Crystal structures of human
CELSR1 EC1-4 and EC4-7 reveal typical folds and a non-canonical linker between EC5 and EC6. Simulations
and experiments using EC4-7 indicate flexibility at EC5-6, and solution experiments show EC7-MAD10-mediated
dimerization. Our results suggest weak homophilic adhesion by CELSR1 cadherin repeats and provide
mechanistic insights into the structural determinants of CELSR1 function.
... 8 Rapid cell proliferation during neural tube closure requires the synthesis of large amounts of nucleotides for DNA replication, 9 while an inadequate supply of nucleotides to neuroepithelial cells might result in neural fold retardation and NTDs. 10 Extensive studies in human and vertebrate models have strongly implicated PCP pathway genes in the etiology of NTDs. 11 It is known that both folate and PCP pathway signaling are involved in cytoskeleton regulation, 12,13 but whether and how folate directly regulates PCP gene expression is unknown. ...
Folate deficiency contribute to neural tube defects (NTDs) which could be rescued by folate supplementation. However, the underlying mechanisms are still not fully understood. Besides, there is considerable controversy concerning the forms of folate used for supplementation. To address this controversy, we prepared culture medium with different forms of folate, folic acid (FA), and 5‐methyltetrahydrofolate (5mTHF), at concentrations of 5 μM, 500 nM, 50 nM, and folate free, respectively. Mouse embryonic fibroblasts (MEFs) were treated with different folates continuously for three passages, and cell proliferation and F‐actin were monitored. We determined that compared to 5mTHF, FA showed stronger effects on promoting cell proliferation and F‐actin formation. We also found that FOLR1 protein level was positively regulated by folate concentration and the non‐canonical Wnt/planar cell polarity (PCP) pathway signaling was significantly enriched among different folate conditions in RNA‐sequencing analyses. We demonstrated for the first time that FOLR1 could promote the transcription of Vangl2, one of PCP core genes. The transcription of Vangl2 was down‐regulated under folate‐deficient condition, which resulted in a decrease in PCP activity and F‐actin formation. In summary, we identified a distinct advantage of FA in cell proliferation and F‐actin formation over 5mTHF, as well as demonstrating that FOLR1 could promote transcription of Vangl2 and provide a new mechanism by which folate deficiency can contribute to the etiology of NTDs.
... The Wnt signaling pathway, which has been known to be associated with several HDAC inhibitors, controls a variety of biological processes, such as cell proliferation, apoptosis, stem cell self-renewal, and differentiation for adult tissue homeostasis and early embryonic development [15][16][17]. This pathway can be classified into the canonical pathway, which leads to transcriptional regulation of cell proliferation, and the non-canonical pathway, which acts on cell polarity and cell migration [16,18,19]. When the canonical Wnt signaling pathway is activated, Wnts (Wnt1, Wnt3a, and Wnt8) bind to the Frizzled (Fzd) receptor attached to low-density lipoprotein receptor-related protein 5/6 (LRP5/6) co-receptors [20]. ...
Histone deacetylase (HDAC) inhibitors promote differentiation through post-translational modifications of histones. BML-281, an HDAC6 inhibitor, has been known to prevent tumors, acute dextran sodium sulfate-associated colitis, and lung injury. However, the neurogenic differentiation effect of BML-281 is poorly understood. In this study, we investigated the effect of BML-281 on neuroblastoma SH-SY5Y cell differentiation into mature neurons by immunocytochemistry (ICC), reverse transcriptase PCR (RT-PCR), quantitative PCR (qPCR), and western blotting analysis. We found that the cells treated with BML-281 showed neurite outgrowth and morphological changes into mature neurons under a microscope. It was confirmed that the gene expression of neuronal markers (NEFL, MAP2, Tuj1, NEFH, and NEFM) was increased with certain concentrations of BML-281. Similarly, the protein expression of neuronal markers (NeuN, Synaptophysin, Tuj1, and NFH) was upregulated with BML-281 compared to untreated cells. Following treatment with BML-281, the expression of Wnt5α increased, and downstream pathways were activated. Interestingly, both Wnt/Ca²⁺ and Wnt/PCP pathways activated and regulated PKC, Cdc42, RhoA, Rac1/2/3, and p-JNK. Therefore, BML-281 induces the differentiation of SH-SY5Y cells into mature neurons by activating the non-canonical Wnt signaling pathway. From these results, we concluded that BML-281 might be a novel drug to differentiation into neuronal cells through the regulation of Wnt signaling pathway to reduce the neuronal cell death.
... Failure to generate coordinated fluid flow in PCP mutant mice leads to left-right patterning defects, defective mucociliary clearance, sterility and hydrocephalus [7,[18][19][20][21][22][23][24]. Core PCP genes are essential for neural tube closure in mammals and pathological variants in PCP genes are strongly associated with neural tube defects in humans [25][26][27]. A notable example of PCP in mammals is the uniform alignment of body hairs across the skin surface, where the core PCP proteins direct polarized morphogenesis and tissue-wide alignment of hair follicles [14,[28][29][30]. ...
Epithelial tissues can be polarized along two axes: in addition to apical-basal polarity they are often also polarized within the plane of the epithelium, known as planar cell polarity (PCP). PCP depends upon the conserved Wnt/Frizzled (Fz) signaling factors, including Fz itself and Van Gogh (Vang/Vangl in mammals). Here, taking advantage of the complementary features of Drosophila wing and mouse skin PCP establishment, we dissect how Vang/Vangl phosphorylation on a specific conserved tyrosine residue affects its interaction with two cytoplasmic core PCP factors, Dishevelled (Dsh/Dvl1-3 in mammals) and Prickle (Pk/Pk1-3). We demonstrate that Pk and Dsh/Dvl bind to Vang/Vangl in an overlapping region centered around this tyrosine. Strikingly, Vang/Vangl phosphorylation promotes its binding to Prickle, a key effector of the Vang/Vangl complex, and inhibits its interaction with Dishevelled. Thus phosphorylation of this tyrosine appears to promote the formation of the mature Vang/Vangl-Pk complex during PCP establishment and conversely it inhibits the Vang interaction with the antagonistic effector Dishevelled. Intriguingly, the phosphorylation state of this tyrosine might thus serve as a switch between transient interactions with Dishevelled and stable formation of Vang-Pk complexes during PCP establishment.
... Previously, we have reported that WDR34 can regulated PCP signaling in vitro (Yin et al., 2020). PCP signaling has been reported to play essential role in neural tube closure through enhancing midline apical constriction and the convergent extension of neural tube (Nikolopoulou et al., 2017;Wang et al., 2019). Therefore, we examined the expression of c-Jun and Juk, two downstream targets of PCP signaling, in both Wdr34 PB/PB and Wdr60 PB/PB embryos by qRT-PCR. ...
Cilia are specialized organelles that extend from plasma membrane, functioning as antennas for signal transduction and are involved in embryonic morphogenesis. Dysfunction of cilia lead to many developmental defects, including neural tube defects (NTDs). Heterodimer WDR60-WDR34 (WD repeat domain 60 and 34) are intermediate chains of motor protein dynein-2, which play important roles in ciliary retrograde transport. It has been reported that disruption of Wdr34 in mouse model results in NTDs and defects of Sonic Hedgehog (SHH) signaling. However, no Wdr60 deficiency mouse model has been reported yet. In this study, piggyBac (PB) transposon is used to interfere Wdr60 and Wdr34 expression respectively to establish Wdr60 PB/PB and Wdr34 PB/PB mouse models. We found that the expression of Wdr60 or Wdr34 is significantly decreased in the homozygote mice. Wdr60 homozygote mice die around E13.5 to E14.5, while Wdr34 homozygote mice die around E10.5 to E11.5. WDR60 is highly expressed in the head region at E10.5 and Wdr60 PB/PB embryos have head malformation. RNAseq and qRT-PCR experiments revealed that Sonic Hedgehog signaling is also downregulated in Wdr60 PB/PB head tissue, demonstrating that WDR60 is also required for promoting SHH signaling. Further experiments on mouse embryos also revealed that the expression levels of planar cell polarity (PCP) components such as CELSR1 and downstream signal molecule c-Jun were downregulated in WDR34 homozygotes compared to wildtype littermates. Coincidently, we observed much higher ratio of open cranial and caudal neural tube in Wdr34 PB/PB mice. CO-IP experiment showed that WDR60 and WDR34 both interact with IFT88, but only WDR34 interacts with IFT140. Taken together, WDR60 and WDR34 play overlapped and distinct functions in modulating neural tube development.
... An intricate balance of WNT signalling is crucial for early forebrain patterning and morphogenesis (Harrison-Uy and Pleasure, 2012;Engelhardt et al., 2022). Impaired WNT signalling leads to severe developmental disorders including neural tube defects (NTDs) (Copp et al., 2003;Wallingford, 2006;Ybot-Gonzalez et al., 2007;Wang et al., 2019;Brown et al., 2020;Zhao et al., 2022). Given that NTDs have a global prevalence of around 19 cases per 10,000 births and are therefore among the most common birth defects (Copp et al., 2003;Harris and Juriloff, 2007;Greene and Copp, 2014;Kakebeen and Niswander, 2021), it is crucial to understand the underlying pathomechanisms. ...
... WNT ligands, such as WNT1 and WNT3a, bind to various types of receptors and co-receptors to transduce signals via the canonical β-catenin-dependent and the non-canonical β-catenin-independent pathways. The most prominent non-canonical WNT pathway is the planar cell polarity (PCP) signalling pathway that regulates cytoskeleton dynamics and collective tissue movements crucial for driving neural tube closure processes (Copp et al., 2003;Wallingford, 2006;Ybot-Gonzalez et al., 2007;Wang et al., 2019). In context of neural tube development, the canonical WNT pathway has mostly been implicated in the regulation of regional identity and in balancing proliferation versus differentiation of early neuronal progenitor cells (NPCs) (Harrison-Uy and Pleasure, 2012;Engelhardt et al., 2022). ...
The specification of the forebrain relies on the precise regulation of WNT/ß-catenin signalling to support neuronal progenitor cell expansion, patterning, and morphogenesis. Imbalances in WNT signalling activity in the early neuroepithelium lead to congenital disorders, such as neural tube defects (NTDs). LDL receptor-related protein (LRP) family members, including the well-studied receptors LRP5 and LRP6, play critical roles in modulating WNT signalling capacity through tightly regulated interactions with their co-receptor Frizzled, WNT ligands, inhibitors and intracellular WNT pathway components. However, little is known about the function of LRP4 as a potential modulator of WNT signalling in the central nervous system. In this study, we investigated the role of LRP4 in the regulation of WNT signalling during early mouse forebrain development. Our results demonstrate that LRP4 can modulate LRP5- and LRP6-mediated WNT signalling in the developing forebrain prior to the onset of neurogenesis at embryonic stage 9.5 and is therefore essential for accurate neural tube morphogenesis. Specifically, LRP4 functions as a genetic modifier for impaired mitotic activity and forebrain hypoplasia, but not for NTDs in LRP6-deficient mutants. In vivo and in vitro data provide evidence that LRP4 is a key player in fine-tuning WNT signalling capacity and mitotic activity of mouse neuronal progenitors and of human retinal pigment epithelial (hTERT RPE-1) cells. Our data demonstrate the crucial roles of LRP4 and LRP6 in regulating WNT signalling and forebrain development and highlight the need to consider the interaction between different signalling pathways to understand the underlying mechanisms of disease. The findings have significant implications for our mechanistic understanding of how LRPs participate in controlling WNT signalling.
... Recent research showed that accumulation of singleton loss-of-function variants contribute to the occurrence of NTDs, regardless of the patient's genetic background or ethnicity (Chen et al., 2018). Many rare loss-of-function variants have been found, which include genes that function in one-carbon metabolism (Marini et al., 2011), and the non-canonical wingless-type MMTV integration site family (WNT)/planar cell polarity, sonic hedgehog, and bone morphogenetic protein (BMP) pathways (Wang et al., 2019). In view of more than 300 genes that are essential in neural tube closure in mice, it is likely that causative genetic factors are largely unknown for human NTDs. ...
Background:
Rare mutations in multiple genes have been associated with human neural tube defects (NTDs), but their causative roles in NTDs disease are poorly understood. Insufficiency of the ribosomal biogenesis gene treacle ribosome biogenesis factor 1(Tcof1) results in cranial NTDs and craniofacial malformations in mice. Here, we aimed to identify genetic association of TCOF1 with human NTDs.
Methods:
High-throughput sequencing targeted on TCOF1 was performed on samples from 355 human cases affected by NTDs and 225 controls from a Han Chinese population.
Results:
Four novel missense variants were found in the NTD cohort. Cell-based assays indicated that the p.(A491G) variant carried by an individual, who shows anencephaly and single-nostril abnormality, attenuates production of total proteins, suggesting a loss-of-function mutation in ribosomal biogenesis. Importantly, this variant promotes nucleolar disruption and stabilizes p53 protein, highlighting an unbalancing effect on cell apoptosis.
Conclusions:
This study explored the functional impact of a missense variant in TCOF1, implicating a set of novel causative biological factors involved in the pathogenicity of human NTDs, particularly whom combined with craniofacial abnormality.
... Y. Yang et al. 15 (53.4%) out of 28 diagnosed cases were attributed to singlegene variants. Therefore, ES and deep WGS, which can detect single-gene variants, should be considered after uninformative clinical karyotyping for these types of anomalies. ...
... The non-canonical Wnt/planar cell polarity (PCP) signaling pathway is implicated in the pathogenesis of NTDs, and 13 genes in the PCP pathway are involved in human NTDs 15 . Rare singleton deleterious variants in these 13 PCP genes (listed in Supplementary Table 3) were identified in two NTD cases, with none found in the control group. ...
Structural anomalies of the central nervous system (CNS) are one of the most common fetal anomalies found during prenatal imaging. However, the genomic architecture of prenatal imaging phenotypes has not yet been systematically studied in a large cohort. Patients diagnosed with fetal CNS anomalies were identified from medical records and images. Fetal samples were subjected to low-pass and deep whole-genome sequencing (WGS) for aneuploid, copy number variation (CNV), single-nucleotide variant (SNV, including insertions/deletions (indels)), and small CNV identification. The clinical significance of variants was interpreted based on a candidate gene list constructed from ultrasound phenotypes. In total, 162 fetuses with 11 common CNS anomalies were enrolled in this study. Primary diagnosis was achieved in 62 cases, with an overall diagnostic rate of 38.3%. Causative variants included 18 aneuploids, 17 CNVs, three small CNVs, and 24 SNVs. Among the 24 SNVs, 15 were novel mutations not reported previously. Furthermore, 29 key genes of diagnostic variants and critical genes of pathogenic CNVs were identified, including five recurrent genes: i.e., TUBA1A , KAT6B , CC2D2A , PDHA1 , and NF1 . Diagnostic variants were present in 34 (70.8%) out of 48 fetuses with both CNS and non-CNS malformations, and in 28 (24.6%) out of 114 fetuses with CNS anomalies only. Hypoplasia of the cerebellum (including the cerebellar vermis) and holoprosencephaly had the highest primary diagnosis yields (>70%), while only four (11.8%) out of 34 neural tube defects achieved genetic diagnosis. Compared with the control group, rare singleton loss-of-function variants (SLoFVs) were significantly accumulated in the patient cohort.
