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01.12.2023 | Original Article

mRNA sequencing provides new insights into the pathogenesis of Hirschsprung’s disease in mice

verfasst von: Qiwen Yang, Fuwen Wang, Zhaofei Wang, Jiajun Guo, Tingjin Chang, Baligen Dalielihan, Ge Yang, Chuzhao Lei, Ruihua Dang

Erschienen in: Pediatric Surgery International | Ausgabe 1/2023

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Abstract

Purpose

The aim of this study is to use RNA sequencing and RT-qPCR to identify the main susceptibility genes linked to the occurrence and development of Hirschsprung disease in the colonic tissues of EDNRB^m1yzcm and wild mice.

Methods

RNA was extracted from colon tissues of 3 mutant homozygous mice and 3 wild mice. RNA degradation, contamination concentration, and integrity were then measured. The extracted RNA was then sequenced using the Illumina platform. The obtained sequence data are filtered to ensure data quality and compared to the reference genome for further analysis. DESeq2 was used for gene expression analysis of the raw data. In addition, graphene oxide enrichment analysis and RT-qPCR validation were also performed.

Results

This study identified 8354 differentially expressed genes in EDNRB^m1yzcm and wild mouse colon tissues by RNA sequencing, including 4346 upregulated genes and 4005 downregulated genes. Correspondingly, the results of RT-qPCR analysis showed good correlation with the transcriptome data. In addition, GO and KEGG enrichment results suggested that there were 8103 terms and 320 pathways in all DEGs. When P < 0.05, 1081 GO terms and 320 KEGG pathways reached a significant level. Finally, through the existing studies and the enrichment results of differentially expressed genes, it was determined that axon guidance and the focal adhesion pathway may be closely related to the occurrence of HSCR.

Conclusions

This study analyzed and identified the differential genes in colonic tissues between EDNRB^m1yzcm mice and wild mice, which provided new insight for further mining the potential pathogenic genes of Hirschsprung’s disease.

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Titel: mRNA sequencing provides new insights into the pathogenesis of Hirschsprung’s disease in mice
verfasst von: Qiwen Yang
Fuwen Wang
Zhaofei Wang
Jiajun Guo
Tingjin Chang
Baligen Dalielihan
Ge Yang
Chuzhao Lei
Ruihua Dang
Publikationsdatum: 01.12.2023
Verlag: Springer Berlin Heidelberg
Erschienen in: Pediatric Surgery International / Ausgabe 1/2023
Print ISSN: 0179-0358
Elektronische ISSN: 1437-9813
DOI: https://doi.org/10.1007/s00383-023-05544-5

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mRNA sequencing provides new insights into the pathogenesis of Hirschsprung’s disease in mice