ORCID
https://orcid.org/0009-0008-1509-5264
Year
2025
Season
Fall
Paper Type
Master's Thesis
College
College of Arts and Sciences
Degree Name
Master of Science in Biology (MS)
Department
Biology
NACO controlled Corporate Body
University of North Florida. Department of Biology
Committee Chairperson
Dr. Marie Mooney
Second Advisor
Dr. David Waddell
Third Advisor
Dr. Judith Ochrietor
Department Chair
Dr. James Gelsleichter
College Dean
Dr. Kaveri Subrahmanyam
Abstract
NGLY1 deficiency is a rare congenital disorder caused by mutations in the NGLY1 gene, which encodes an enzyme responsible for deglycosylating misfolded glycoproteins prior to proteasomal degradation. Affected individuals exhibit severe neurological and developmental impairments, including intellectual disability, movement disorder accompanied by severely diminished tear production (Alacrima), increased liver transaminases, and seizures. Despite extensive research on pathways disrupted by NGLY1 loss, the role of Wnt signaling, a critical regulator of neurodevelopment, remains unexplored. Here, we investigated whether NGLY1 deficiency alters Wnt pathway activity in mouse embryonic fibroblasts (MEFs). Transcriptomic analysis revealed downregulation of Wnt target genes and reduced β-catenin, a protein critical for the Wnt signaling pathway, in NGLY1-deficient MEFs. To test reversibility, we treated mutant and wild-type MEFs with a Wnt agonist and assessed β-catenin localization and Wnt target gene expression using RT-qPCR and immunofluorescence imaging. Our results demonstrated that pharmacological inhibition of GSK3, a key negative regulator of Wnt signaling, using CHIR99021, significantly increased cell viability, promoted β-catenin accumulation in both cytoplasmic and nuclear compartments, and upregulated the expression of Wnt target genes in NGLY1-deficient mouse embryonic fibroblasts. These findings identify Wnt signaling suppression as a novel consequence of NGLY1 deficiency and highlight GSK3 inhibition as a potential therapeutic strategy to restore Wnt pathway activity and mitigate cellular phenotypes associated with this disorder.
Suggested Citation
Ghannad Zadeh, Rojin, "Targeting Wnt signaling with GSK3 inhibition restores pathway activity in NGLY1-deficient mouse embryonic fibroblasts" (2025). UNF Graduate Theses and Dissertations. 1381.
https://digitalcommons.unf.edu/etd/1381
Included in
Cell Biology Commons, Congenital, Hereditary, and Neonatal Diseases and Abnormalities Commons, Developmental Biology Commons, Molecular and Cellular Neuroscience Commons, Nervous System Diseases Commons