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Published on: December 16, 2016
The Metabolic Reprogramming of Frem2 Mutant Mice Embryos in Cryptophthalmos Development
Xiayin Zhang1, Ruixin Wang1, Ting Wang1
1State Key Laboratory of Ophthalmology, Zhongshan Ophthalmic Center, Sun Yat-sen University, Guangzhou, China.
Background:
Cryptophthalmos is characterized by congenital ocular dysplasia with eyelid malformation. The pathogenicity of mutations in genes encoding components of the FRAS1/FREM protein complex is well established, but the underlying pathomechanisms of this disease are still unclear. In the previous study, we generated mice carrying Frem2 compound heterozygous mutations using CRISPR/Cas9 and showed that these mice recapitulated the human cryptophthalmos phenotype.
Methods:
In this study, we tracked changes in the metabolic profile of embryos and expression of metabolism-related genes in Frem2 mutant mice on E13.5 compared with wild-type mice. RNA sequencing (RNA-seq) was utilized to decipher the differentiated expression of genes associated with metabolism. Untargeted metabolomics and targeted metabolomics analyses were performed to detect and verify the shifts in the composition of the embryonic metabolome.
Results:
Differentially expressed genes participating in amino acid metabolism and energy metabolism were observed by RNA-seq. Transcriptomic analysis suggests that 821 (39.89%) up-regulated genes and 320 (32.99%) down-regulated genes were involved in the metabolic process in the enriched GO terms. A total of 92 significantly different metabolites were identified including creatine, guanosine 5'-monophosphate, cytosine, cytidine 5'-monophosphate, adenine, and L-serine. Interestingly, major shifts related to ATP binding cassette transporters (ABC transporters) and the biosynthesis of amino acids in the composition of the embryonic metabolome were observed by KEGG metabolic analysis, indicating that these pathways could also be involved in the pathogenesis of cryptophthalmos.
Conclusion:
We demonstrate that Frem2 mutant fetal mice have increased susceptibility to the disruption of eye morphogenesis in association with distinct transcriptomic and metabolomic signatures. Our findings suggest that the metabolomic signature established before birth may play a role in mediating cryptophthalmos in Frem2 mutant mice, which may have important implications for the pathogenesis of cryptophthalmos.
Insights
Mutations in Frem2 disrupt eye development in mice, leading to cryptophthalmos. Metabolic changes in embryos, including amino acid and energy metabolism shifts, are linked to this congenital condition.
Area of Science:
- Developmental Biology
- Genetics
- Metabolomics
Background:
- Cryptophthalmos is a congenital disorder characterized by ocular dysplasia and eyelid malformations.
- Pathogenic mutations in genes encoding the FRAS1/FREM protein complex are known, but disease mechanisms remain unclear.
- Previous studies established a mouse model for cryptophthalmos using CRISPR/Cas9 to induce compound heterozygous mutations in Frem2.
Purpose of the Study:
- To investigate the metabolic and transcriptomic changes in Frem2 mutant mouse embryos.
- To elucidate the pathomechanisms underlying cryptophthalmos in Frem2 mutant mice.
Main Methods:
- RNA sequencing (RNA-seq) to analyze differential gene expression related to metabolism.
- Untargeted and targeted metabolomics to identify shifts in embryonic metabolite composition.
- KEGG pathway analysis to understand metabolic alterations.
Main Results:
- RNA-seq revealed differential expression of genes involved in amino acid and energy metabolism.
- 92 significantly different metabolites were identified, including creatine, guanosine 5'-monophosphate, and L-serine.
- KEGG analysis indicated major shifts in ATP binding cassette transporters and amino acid biosynthesis pathways.
Conclusions:
- Frem2 mutant fetal mice exhibit distinct transcriptomic and metabolomic signatures associated with disrupted eye morphogenesis.
- Pre-natal metabolomic signatures may play a crucial role in mediating cryptophthalmos pathogenesis in Frem2 mutant mice.
- These findings offer insights into the broader pathogenesis of cryptophthalmos.
