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GAL3ST1 Deficiency Reduces Epithelial-Mesenchymal Transition and Tumorigenic Capacity in a Cholangiocarcinoma Cell
Lin Chen1, Montserrat Elizalde2, Ludwig J Dubois3
1Department of Surgery, School of Nutrition and Translational Research in Metabolism, Maastricht University, 6200 MD Maastricht, The Netherlands.
Abstract:
Cholangiocarcinoma (CCA), or bile duct cancer, is the second most common liver malignancy, with an increasing incidence in Western countries. The lack of effective treatments associated with the absence of early symptoms highlights the need to search for new therapeutic targets for CCA. Sulfatides (STs), a type of sulfoglycosphingolipids, have been found in the biliary tract, with increased levels in CCA and other types of cancer. STs are involved in protein trafficking and cell adhesion as part of the lipid rafts of the plasma membrane. We aimed to study the role of STs in CCA by the genetic targeting of GAL3ST1, an enzyme involved in ST synthesis. We used the CRISPR-Cas9 system to generate GAL3ST1-deficient TFK1 cells. GAL3ST1 KO cells showed lower proliferation and clonogenic activity and reduced glycolytic activity compared to TFK1 cells. Polarized TFK1 GAL3ST1 KO cells displayed increased transepithelial resistance and reduced permeability compared to TFK1 wt cells. The loss of GAL3ST1 showed a negative effect on growth in 30 out of 34 biliary tract cancer cell lines from the DepMap database. GAL3ST1 deficiency partially restored epithelial identity and barrier function and reduced proliferative activity in CCA cells. Sulfatide synthesis may provide a novel therapeutic target for CCA.
Insights
Targeting sulfatide synthesis may offer a new therapeutic strategy for cholangiocarcinoma (bile duct cancer). Inhibiting GAL3ST1 reduced cancer cell growth and improved barrier function, suggesting a potential treatment pathway.
Area of Science:
- Oncology
- Biochemistry
- Molecular Biology
Background:
- Cholangiocarcinoma (CCA) is a prevalent liver malignancy with limited treatment options.
- Elevated sulfatides (STs), a type of sulfoglycosphingolipid, are observed in CCA and other cancers.
- STs play roles in cellular processes like protein trafficking and cell adhesion within lipid rafts.
Purpose of the Study:
- To investigate the role of STs in CCA by genetically targeting GAL3ST1, an enzyme crucial for ST synthesis.
- To evaluate the impact of GAL3ST1 deficiency on CCA cell behavior and characteristics.
Main Methods:
- CRISPR-Cas9 gene editing was employed to create GAL3ST1-deficient (KO) TFK1 cells.
- Proliferation, clonogenic activity, glycolytic activity, and barrier function (transepithelial resistance and permeability) were assessed.
- The effect of GAL3ST1 loss on growth was analyzed across multiple biliary tract cancer cell lines using DepMap database data.
Main Results:
- GAL3ST1 KO cells exhibited reduced proliferation, clonogenic, and glycolytic activity compared to wild-type (wt) cells.
- Polarized GAL3ST1 KO cells demonstrated enhanced barrier function with increased transepithelial resistance and decreased permeability.
- GAL3ST1 deficiency negatively impacted growth in a majority (30/34) of tested biliary tract cancer cell lines.
Conclusions:
- GAL3ST1 deficiency partially restored epithelial identity and barrier function in CCA cells.
- Reduced proliferative activity was observed in CCA cells with GAL3ST1 deficiency.
- Sulfatide synthesis represents a potential novel therapeutic target for cholangiocarcinoma treatment.
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