Department of Reproductive Medicine Center, Henan Provincial People's Hospital, People's Hospital of Zhengzhou University, Zhengzhou, Henan Province, People's Republic of China.
Center for Precision Environmental Health, Department of Molecular and Cellular Biology, Baylor College of Medicine, Houston, Texas, USA.
FASEB J. 2024 Jan;38(1):e23346. doi: 10.1096/fj.202300202R.
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.
叶酸缺乏可导致神经管缺陷(NTDs),而叶酸补充可以挽救这一缺陷。然而,其潜在机制尚未完全阐明。此外,用于补充的叶酸形式存在相当大的争议。为了解决这一争议,我们分别用叶酸、叶酸酸(FA)和 5-甲基四氢叶酸(5mTHF)这三种不同形式的叶酸制备了浓度分别为 5μM、500nM、50nM 和无叶酸的培养基。我们用不同的叶酸连续处理小鼠胚胎成纤维细胞(MEFs)三个传代,监测细胞增殖和 F-肌动蛋白。我们发现与 5mTHF 相比,FA 对促进细胞增殖和 F-肌动蛋白形成具有更强的作用。我们还发现 FOLR1 蛋白水平受叶酸浓度的正向调节,并且在 RNA 测序分析中,非经典 Wnt/平面细胞极性(PCP)信号通路在不同叶酸条件下显著富集。我们首次证明 FOLR1 可以促进 PCP 核心基因之一 Vangl2 的转录。在叶酸缺乏条件下,Vangl2 的转录下调,导致 PCP 活性和 F-肌动蛋白形成减少。总之,我们确定了 FA 在细胞增殖和 F-肌动蛋白形成方面相对于 5mTHF 的明显优势,并证明了 FOLR1 可以促进 Vangl2 的转录,并提供了一种新的机制,即叶酸缺乏可导致 NTDs 的发生。