Inpp5e Regulated the Cilium-Related Genes Contributing to the Neural Tube Defects Under 5-Fluorouracil Exposure

Xiuwei Wang; Jialu Yu; Huixuan Yue; Shen Li; Aiyun Yang; Zhiqiang Zhu; Zhen Guan; Jianhua Wang

doi:10.1007/s12035-024-03946-7

Inpp5e Regulated the Cilium-Related Genes Contributing to the Neural Tube Defects Under 5-Fluorouracil Exposure

Mol Neurobiol. 2024 Jan 29. doi: 10.1007/s12035-024-03946-7. Online ahead of print.

Authors

Xiuwei Wang^#¹, Jialu Yu^#^{1

2}, Huixuan Yue³, Shen Li¹, Aiyun Yang¹, Zhiqiang Zhu¹, Zhen Guan⁴, Jianhua Wang^{5

6}

Affiliations

¹ Laboratory of Translational Medicine, Beijing Municipal Key Laboratory of Child Development and Nutriomics, Capital Institute of Pediatrics, Beijing, 100020, China.
² Graduate School of Peking Union Medical College, Chinese Academy of Medical Sciences, Beijing, 100730, China.
³ Department of Pediatrics, Beijing Chaoyang Hospital of Capital Medical University, Beijing, 100020, China.
⁴ Laboratory of Translational Medicine, Beijing Municipal Key Laboratory of Child Development and Nutriomics, Capital Institute of Pediatrics, Beijing, 100020, China. guanzhen@aliyun.com.
⁵ Laboratory of Translational Medicine, Beijing Municipal Key Laboratory of Child Development and Nutriomics, Capital Institute of Pediatrics, Beijing, 100020, China. fywjh@163.com.
⁶ Graduate School of Peking Union Medical College, Chinese Academy of Medical Sciences, Beijing, 100730, China. fywjh@163.com.

^# Contributed equally.

PMID: 38285286
DOI: 10.1007/s12035-024-03946-7

Abstract

Primary cilia are crucial for neurogenesis, and cilium-related genes are involved in the closure of neural tubes. Inositol polyphosphate-5-phosphatase (Inpp5e) was enriched in primary cilia and closely related to the occurrence of neural tube defects (NTDs). However, the role of Inpp5e in the development of NTDs is not well-known. To investigate whether Inpp5e gene is associated with the neural tube closure, we established a mouse model of NTDs by 5-fluorouracil (5-FU) exposure at gestational day 7.5 (GD7.5). The Inpp5e knockdown (Inpp5e^-/-) mouse embryonic stem cells (mESCs) were produced by CRISPR/Cas9 system. The expressions of Inpp5e and other cilium-related genes including intraflagellar transport 80 (Ift80), McKusick-Kaufman syndrome (Mkks), and Kirsten rat sarcoma viral oncogene homolog (Kras) were determined, utilizing quantitative real-time reverse transcription polymerase chain reaction (qRT-PCR), western blot, PCR array, and immunofluorescence staining. The result showed that the incidence of NTDs was 37.10% (23 NTDs/62 total embryos) and significantly higher than that in the control group (P < 0.001). The neuroepithelial cells of neural tubes were obviously disarranged in NTD embryos. The mRNA and protein levels of Inpp5e, Ift80, Mkks, and Kras were significantly decreased in NTD embryonic brain tissues, compared to the control (P < 0.05). Knockdown of the Inpp5e (Inpp5e^-/-) reduced the expressions of Ift80, Mkks, and Kras in mESCs. Furthermore, the levels of α-tubulin were significantly reduced in NTD embryonic neural tissue and Inpp5e^-/- mESCs. These results suggested that maternal 5-FU exposure inhibited the expression of Inpp5e, which resulted in the downregulation of cilium-related genes (Ift80, Mkks, and Kras), leading to the impairment of primary cilium development, and ultimately disrupted the neural tube closure.

Keywords: 5-fluorouracil; Inpp5e; Neural tube defects; Primary cilia.

Abstract

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