Deacetylated Sialic Acid Sensitizes Lung and Colon Cancers to Novel Cucurbitacin-Inspired Estrone Epidermal Growth Factor Receptor (EGFR) Inhibitor Analogs
Cancers utilize sugar residues such as sialic acids (Sia) to improve their ability to survive. Sia presents a variety of functional group alterations, including O-acetylation on the C6 hydroxylated tail. Previously, sialylation has been reported to suppress EGFR activation and increase cancer cell s...
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| Format: | Article |
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MDPI AG
2023-08-01
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| Series: | Molecules |
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| Online Access: | https://www.mdpi.com/1420-3049/28/17/6257 |
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| author | Mathias T. Anim Isaac Tuffour Rylan Willis Matthew Schell Trevor Ostlund Mater H. Mahnashi Fathi Halaweish Rachel Willand-Charnley |
| author_facet | Mathias T. Anim Isaac Tuffour Rylan Willis Matthew Schell Trevor Ostlund Mater H. Mahnashi Fathi Halaweish Rachel Willand-Charnley |
| author_sort | Mathias T. Anim |
| collection | DOAJ |
| description | Cancers utilize sugar residues such as sialic acids (Sia) to improve their ability to survive. Sia presents a variety of functional group alterations, including O-acetylation on the C6 hydroxylated tail. Previously, sialylation has been reported to suppress EGFR activation and increase cancer cell sensitivity to Tyrosine Kinase Inhibitors (TKIs). In this study, we report on the effect of deacetylated Sia on the activity of three novel EGFR-targeting Cucurbitacin-inspired estrone analogs (CIEAs), MMA 294, MMA 321, and MMA 320, in lung and colon cancer cells. Acetylation was modulated by the removal of Sialate O-Acetyltransferase, also known as CAS1 Domain-containing protein (CASD1) gene via CRISPR-Cas9 gene editing. Using a variety of cell-based approaches including MTT cell viability assay, flow cytometry, immunofluorescence assay and in-cell ELISA we observed that deacetylated Sia-expressing knockout cells (1.24–6.49 μM) were highly sensitive to all CIEAs compared with the control cells (8.82–20.97 μM). Apoptosis and varied stage cell cycle arrest (G0/G1 and G2/M) were elucidated as mechanistic modes of action of the CIEAs. Further studies implicated overexpression of CIEAs’ cognate protein target, phosphorylated EGFR, in the chemosensitivity of the deacetylated Sia-expressing knockout cells. This observation correlated with significantly decreased levels of key downstream proteins (phosphorylated ERK and mTOR) of the EGFR pathway in knockout cells compared with controls when treated with CIEAs. Collectively, our findings indicate that Sia deacetylation renders lung and colon cancer cells susceptible to EGFR therapeutics and provide insights for future therapeutic interventions. |
| first_indexed | 2024-03-10T23:17:34Z |
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| issn | 1420-3049 |
| language | English |
| last_indexed | 2024-03-10T23:17:34Z |
| publishDate | 2023-08-01 |
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| spelling | doaj.art-988cc52959dc4889a0680ffb96549a3a2023-11-19T08:33:28ZengMDPI AGMolecules1420-30492023-08-012817625710.3390/molecules28176257Deacetylated Sialic Acid Sensitizes Lung and Colon Cancers to Novel Cucurbitacin-Inspired Estrone Epidermal Growth Factor Receptor (EGFR) Inhibitor AnalogsMathias T. Anim0Isaac Tuffour1Rylan Willis2Matthew Schell3Trevor Ostlund4Mater H. Mahnashi5Fathi Halaweish6Rachel Willand-Charnley7Department of Chemistry and Biochemistry, South Dakota State University, Brookings, SD 57007, USADepartment of Chemistry and Biochemistry, South Dakota State University, Brookings, SD 57007, USADepartment of Chemistry and Biochemistry, South Dakota State University, Brookings, SD 57007, USADepartment of Chemistry and Biochemistry, South Dakota State University, Brookings, SD 57007, USADepartment of Chemistry and Biochemistry, South Dakota State University, Brookings, SD 57007, USADepartment of Pharmaceutical Chemistry, Najran University, Najran P.O. Box 1988, Saudi ArabiaDepartment of Chemistry and Biochemistry, South Dakota State University, Brookings, SD 57007, USADepartment of Chemistry and Biochemistry, South Dakota State University, Brookings, SD 57007, USACancers utilize sugar residues such as sialic acids (Sia) to improve their ability to survive. Sia presents a variety of functional group alterations, including O-acetylation on the C6 hydroxylated tail. Previously, sialylation has been reported to suppress EGFR activation and increase cancer cell sensitivity to Tyrosine Kinase Inhibitors (TKIs). In this study, we report on the effect of deacetylated Sia on the activity of three novel EGFR-targeting Cucurbitacin-inspired estrone analogs (CIEAs), MMA 294, MMA 321, and MMA 320, in lung and colon cancer cells. Acetylation was modulated by the removal of Sialate O-Acetyltransferase, also known as CAS1 Domain-containing protein (CASD1) gene via CRISPR-Cas9 gene editing. Using a variety of cell-based approaches including MTT cell viability assay, flow cytometry, immunofluorescence assay and in-cell ELISA we observed that deacetylated Sia-expressing knockout cells (1.24–6.49 μM) were highly sensitive to all CIEAs compared with the control cells (8.82–20.97 μM). Apoptosis and varied stage cell cycle arrest (G0/G1 and G2/M) were elucidated as mechanistic modes of action of the CIEAs. Further studies implicated overexpression of CIEAs’ cognate protein target, phosphorylated EGFR, in the chemosensitivity of the deacetylated Sia-expressing knockout cells. This observation correlated with significantly decreased levels of key downstream proteins (phosphorylated ERK and mTOR) of the EGFR pathway in knockout cells compared with controls when treated with CIEAs. Collectively, our findings indicate that Sia deacetylation renders lung and colon cancer cells susceptible to EGFR therapeutics and provide insights for future therapeutic interventions.https://www.mdpi.com/1420-3049/28/17/6257sialic acidCASD1apoptosisestrone analogsEpidermal Growth Factor Receptor (EGFR) |
| spellingShingle | Mathias T. Anim Isaac Tuffour Rylan Willis Matthew Schell Trevor Ostlund Mater H. Mahnashi Fathi Halaweish Rachel Willand-Charnley Deacetylated Sialic Acid Sensitizes Lung and Colon Cancers to Novel Cucurbitacin-Inspired Estrone Epidermal Growth Factor Receptor (EGFR) Inhibitor Analogs Molecules sialic acid CASD1 apoptosis estrone analogs Epidermal Growth Factor Receptor (EGFR) |
| title | Deacetylated Sialic Acid Sensitizes Lung and Colon Cancers to Novel Cucurbitacin-Inspired Estrone Epidermal Growth Factor Receptor (EGFR) Inhibitor Analogs |
| title_full | Deacetylated Sialic Acid Sensitizes Lung and Colon Cancers to Novel Cucurbitacin-Inspired Estrone Epidermal Growth Factor Receptor (EGFR) Inhibitor Analogs |
| title_fullStr | Deacetylated Sialic Acid Sensitizes Lung and Colon Cancers to Novel Cucurbitacin-Inspired Estrone Epidermal Growth Factor Receptor (EGFR) Inhibitor Analogs |
| title_full_unstemmed | Deacetylated Sialic Acid Sensitizes Lung and Colon Cancers to Novel Cucurbitacin-Inspired Estrone Epidermal Growth Factor Receptor (EGFR) Inhibitor Analogs |
| title_short | Deacetylated Sialic Acid Sensitizes Lung and Colon Cancers to Novel Cucurbitacin-Inspired Estrone Epidermal Growth Factor Receptor (EGFR) Inhibitor Analogs |
| title_sort | deacetylated sialic acid sensitizes lung and colon cancers to novel cucurbitacin inspired estrone epidermal growth factor receptor egfr inhibitor analogs |
| topic | sialic acid CASD1 apoptosis estrone analogs Epidermal Growth Factor Receptor (EGFR) |
| url | https://www.mdpi.com/1420-3049/28/17/6257 |
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