Journal of Shanghai Jiao Tong University (Medical Science) >

2024 , Vol. 44 >Issue 7: 871 - 882

DOI: https://doi.org/10.3969/j.issn.1674-8115.2024.07.008

Basic research

Study on the developmental function of CT14 using the model organism Caenorhabditis elegans

  • Shuwen YANG ,
  • Juan CHEN ,
  • Qin YANG ,
  • Ming LEI ,
  • Chenhui HUANG
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  • Shanghai Institute of Precision Medicine, Shanghai Ninth People′s Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200125, China
LEI Ming, E-mail: Leim@shsmu.edu.cn.
HUANG Chenhui, E-mail: huangchh@shsmu.edu.cn

Received date: 2024-02-27

  Accepted date: 2024-03-21

  Online published: 2024-07-28

Supported by

National Natural Science Foundation of China(31971137)

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Abstract

Objective ·To investigate the effects of the cancer-testis antigen 14 (CT14) on embryonic and larval development in nematodes by using the model organism Caenorhabditis elegans (C. elegans), aiming to uncover its potential functions and mechanisms during development. Methods ·Transgenic C. elegans strains were constructed by using microinjection for the inducible expression of human CT14 (HsCT14), a truncated mutant of CT14 (HsCT14?CIR) lacking CT14-specific intermediate region (CIR), and a green fluorescent protein (GFP) control. The impacts of full-length and truncated mutant CT14 on nematode embryonic and larval development were analyzed and compared. Additionally, transgenic C. elegans strains with inducible expression of CT14 from various primates, including the crab-eating macaque (Macaca fascicularis) and mouse lemur (Microcebus murinus), were also constructed to assess the effects on egg hatching and larval-to-adult transformation rates. The differential gene expression in nematode embryos induced by CT14 was analyzed by Smart-seq transcriptome sequencing, with further insights gained through KEGG (Kyoto Encyclopedia of Genes and Genomes) and GSEA (Gene Set Enrichment Analysis), to explore the involved biological processes and pathways. Results ·The induced expression of HsCT14 and its truncated mutant HsCT14?CIR significantly reduced the hatching rate of nematode eggs, with a more pronounced effect observed in HsCT14-expressing strains. Differential interference contrast (DIC) microscopy imaging revealed significant morphological abnormalities in embryos expressing HsCT14 during the comma stage. Nematodes expressing HsCT14 or HsCT14?CIR exhibited developmental arrest in larvae and substantially lower larval-to-adult transformation rates compared to the GFP control. The impact was more pronounced in nematodes expressing HsCT14 than those with HsCT14?CIR. The expression of Macaca fascicularis CT14 (MfCT14) exhibited significant effects on the hatching rate and adult transformation rate, similar to that of HsCT14, while the expression of Microcebus murinus CT14 (MmCT14) displayed significantly reduced impact compared to HsCT14 and MfCT14. Smart-seq results indicated that CT14 expression affected various biological processes in nematode embryos, related to ATP-dependent chromatin remodeling and DNA replication. Conclusion ·Ectopic expression of the cancer-testis antigen CT14 significantly disrupts both embryonic and larval developments in C. elegans, with the CIR sequence substantially enhancing this effect. It suggests that CT14 may play an important regulatory role in biological development by affecting gene expression in multiple pathways, including chromatin remodeling.

Key words： cancer-testis antigen; cancer-testis antigen 14 (CT14); Caenorhabditis elegans; embryonic development; larval development

Cite this article

Shuwen YANG , Juan CHEN , Qin YANG , Ming LEI , Chenhui HUANG . Study on the developmental function of CT14 using the model organism Caenorhabditis elegans[J]. Journal of Shanghai Jiao Tong University (Medical Science), 2024 , 44(7) : 871 -882 . DOI: 10.3969/j.issn.1674-8115.2024.07.008

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