Wnt signaling pathways, including the canonical Wnt/β-catenin pathway, planar cell polarity pathway, and Wnt/Ca²⁺ signaling pathway, play important roles in neural development during embryonic stages. The DVL genes encode the hub proteins for Wnt signaling pathways. The mutations in DVL2 and DVL3 were identified from patients with neural tube defects (NTDs), but their functions in the pathogenesis of human neural diseases remain elusive. Here, we sequenced the coding regions of three DVL genes in 176 stillborn or miscarried fetuses with NTDs or Dandy-Walker malformation (DWM) and 480 adult controls from a Han Chinese population. Four rare mutations were identified: DVL1 p.R558H, DVL1 p.R606C, DVL2 p.R633W, and DVL3 p.R222Q. To assess the effect of these mutations on NTDs and DWM, various functional analyses such as luciferase reporter assay, stress fiber formation, and in vivo teratogenic assay were performed. The results showed that the DVL2 p.R633W mutation destabilized DVL2 protein and upregulated activities for all three Wnt signalings (Wnt/β-catenin signaling, Wnt/planar cell polarity signaling, and Wnt/Ca²⁺ signaling) in mammalian cells. In contrast, DVL1 mutants (DVL1 p.R558H and DVL1 p.R606C) decreased canonical Wnt/β-catenin signaling but increased the activity of Wnt/Ca²⁺ signaling, and DVL3 p.R222Q only decreased the activity of Wnt/Ca²⁺ signaling. We also found that only the DVL2 p.R633W mutant displayed more severe teratogenicity in zebrafish embryos than wild-type DVL2. Our study demonstrates that these four rare DVL mutations, especially DVL2 p.R633W, may contribute to human neural diseases such as NTDs and DWM by obstructing Wnt signaling pathways.

Wnt信号传导通路，包括规范的Wnt /β-catenin通路，平面细胞极性通路和Wnt / Ca ²⁺信号通路，在胚胎期的神经发育中起着重要作用。的DVL基因编码Wnt信号途径中的蛋白质毂。从具有神经管缺陷（NTD）的患者中鉴定出DVL2和DVL3的突变，但它们在人类神经疾病发病机理中的功能仍然难以捉摸。在这里，我们对176例NTDs或Dandy-Walker畸形（DWM）的死胎或流产胎儿和480名汉族成年人的三个DVL基因的编码区进行了测序。确定了四个罕见突变：DVL1 p.R558H，DVL1 p.R606C，DVL2 p.R633W和DVL3 p.R222Q。为了评估这些突变对NTD和DWM的影响，进行了各种功能分析，例如荧光素酶报告基因分析，应激纤维形成和体内致畸分析。结果表明，哺乳动物细胞中所有三种Wnt信号（Wnt /β-catenin信号，Wnt /平面细胞极性信号和Wnt / Ca ^{2 +}信号）的DVL2 p.R633W突变使DVL2蛋白不稳定，并上调活性。相比之下，DVL1突变体（DVL1 p.R558H和DVL1p.R606C）下降经典Wnt /β-catenin信号但增加的Wnt / Ca的活性^{2 +}信令，且DVL 3 p.R222Q仅下降的Wnt /活性的Ca ^{2 +}信令。我们还发现，仅DVL2 p.R633W突变体在斑马鱼胚胎中显示出比野生型DVL2更严重的致畸性。我们的研究表明，这四个罕见的DVL突变，特别是DVL2 p.R633W，可能通过阻碍Wnt信号通路而导致人类神经疾病，例如NTD和DWM。