Gb3‐cSrc complex in glycosphingolipid‐enriched microdomains contributes to the expression of p53 mutant protein and cancer drug resistance via β‐catenin–activated RNA methylation
Abstract Glucosylceramide synthase (GCS) is a key enzyme catalyzing ceramide glycosylation to generate glucosylceramide (GlcCer), which in turn serves as the precursor for cells to produce glycosphingolipids (GSLs). In cell membranes, GSLs serve as essential components of GSL‐enriched microdomains (...
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| Format: | Article |
| Language: | English |
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Wiley
2020-11-01
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| Series: | FASEB BioAdvances |
| Subjects: | |
| Online Access: | https://doi.org/10.1096/fba.2020-00044 |
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| author | Kartik R. Roy Mohammad B. Uddin Sagor C. Roy Ronald A. Hill John Marshall Yu‐Teh Li Jean Christopher Chamcheu Hua Lu Yong‐Yu Liu |
| author_facet | Kartik R. Roy Mohammad B. Uddin Sagor C. Roy Ronald A. Hill John Marshall Yu‐Teh Li Jean Christopher Chamcheu Hua Lu Yong‐Yu Liu |
| author_sort | Kartik R. Roy |
| collection | DOAJ |
| description | Abstract Glucosylceramide synthase (GCS) is a key enzyme catalyzing ceramide glycosylation to generate glucosylceramide (GlcCer), which in turn serves as the precursor for cells to produce glycosphingolipids (GSLs). In cell membranes, GSLs serve as essential components of GSL‐enriched microdomains (GEMs) and mediate membrane functions and cell behaviors. Previous studies showed that ceramide glycosylation correlates with upregulated expression of p53 hotspot mutant R273H and cancer drug resistance. Yet, the underlying mechanisms remain elusive. We report herewith that globotriaosylceramide (Gb3) is associated with cSrc kinase in GEMs and plays a crucial role in modulating expression of p53 R273H mutant and drug resistance. Colon cancer cell lines, either WiDr homozygous for missense‐mutated TP53 (R273H+/+) or SW48/TP53‐Dox bearing heterozygous TP53 mutant (R273H/+), display drug resistance with increased ceramide glycosylation. Inhibition of GCS with Genz‐161 (GENZ 667161) resensitized cells to apoptosis in these p53 mutant‐carrying cancer cells. Genz‐161 effectively inhibited GCS activity, and substantially suppressed the elevated Gb3 levels seen in GEMs of p53‐mutant cells exposed to doxorubicin. Complex formation between Gb3 and cSrc in GEMs to activate β‐catenin was detected in both cultured cells and xenograft tumors. Suppression of ceramide glycosylation significantly decreased Gb3‐cSrc in GEMs, β‐catenin, and methyltransferase‐like 3 for m6A RNA methylation, thus altering pre‐mRNA splicing, resulting in upregulated expression of wild‐type p53 protein, but not mutants, in cells carrying p53 R273H. Altogether, increased Gb3‐cSrc complex in GEMs of membranes in response to anticancer drug induced cell stress promotes expression of p53 mutant proteins and accordant cancer drug resistance. |
| first_indexed | 2024-12-12T14:25:40Z |
| format | Article |
| id | doaj.art-973d41d7eb66424a8dba340670718eb5 |
| institution | Directory Open Access Journal |
| issn | 2573-9832 |
| language | English |
| last_indexed | 2024-12-12T14:25:40Z |
| publishDate | 2020-11-01 |
| publisher | Wiley |
| record_format | Article |
| series | FASEB BioAdvances |
| spelling | doaj.art-973d41d7eb66424a8dba340670718eb52022-12-22T00:21:41ZengWileyFASEB BioAdvances2573-98322020-11-0121165366710.1096/fba.2020-00044Gb3‐cSrc complex in glycosphingolipid‐enriched microdomains contributes to the expression of p53 mutant protein and cancer drug resistance via β‐catenin–activated RNA methylationKartik R. Roy0Mohammad B. Uddin1Sagor C. Roy2Ronald A. Hill3John Marshall4Yu‐Teh Li5Jean Christopher Chamcheu6Hua Lu7Yong‐Yu Liu8School of Basic Pharmaceutical and Toxicological Sciences College of Pharmacy University of Louisiana at Monroe Monroe Louisiana USASchool of Basic Pharmaceutical and Toxicological Sciences College of Pharmacy University of Louisiana at Monroe Monroe Louisiana USASchool of Basic Pharmaceutical and Toxicological Sciences College of Pharmacy University of Louisiana at Monroe Monroe Louisiana USASchool of Basic Pharmaceutical and Toxicological Sciences College of Pharmacy University of Louisiana at Monroe Monroe Louisiana USADepartment of Rare Genetic Disease Research Sanofi‐Genzyme R&D Center Genzyme, Framingham Massachusetts USADepartment of Biochemistry and Molecular Biology Tulane University School of Medicine New Orleans Louisiana USASchool of Basic Pharmaceutical and Toxicological Sciences College of Pharmacy University of Louisiana at Monroe Monroe Louisiana USADepartment of Biochemistry and Molecular Biology Tulane University School of Medicine New Orleans Louisiana USASchool of Basic Pharmaceutical and Toxicological Sciences College of Pharmacy University of Louisiana at Monroe Monroe Louisiana USAAbstract Glucosylceramide synthase (GCS) is a key enzyme catalyzing ceramide glycosylation to generate glucosylceramide (GlcCer), which in turn serves as the precursor for cells to produce glycosphingolipids (GSLs). In cell membranes, GSLs serve as essential components of GSL‐enriched microdomains (GEMs) and mediate membrane functions and cell behaviors. Previous studies showed that ceramide glycosylation correlates with upregulated expression of p53 hotspot mutant R273H and cancer drug resistance. Yet, the underlying mechanisms remain elusive. We report herewith that globotriaosylceramide (Gb3) is associated with cSrc kinase in GEMs and plays a crucial role in modulating expression of p53 R273H mutant and drug resistance. Colon cancer cell lines, either WiDr homozygous for missense‐mutated TP53 (R273H+/+) or SW48/TP53‐Dox bearing heterozygous TP53 mutant (R273H/+), display drug resistance with increased ceramide glycosylation. Inhibition of GCS with Genz‐161 (GENZ 667161) resensitized cells to apoptosis in these p53 mutant‐carrying cancer cells. Genz‐161 effectively inhibited GCS activity, and substantially suppressed the elevated Gb3 levels seen in GEMs of p53‐mutant cells exposed to doxorubicin. Complex formation between Gb3 and cSrc in GEMs to activate β‐catenin was detected in both cultured cells and xenograft tumors. Suppression of ceramide glycosylation significantly decreased Gb3‐cSrc in GEMs, β‐catenin, and methyltransferase‐like 3 for m6A RNA methylation, thus altering pre‐mRNA splicing, resulting in upregulated expression of wild‐type p53 protein, but not mutants, in cells carrying p53 R273H. Altogether, increased Gb3‐cSrc complex in GEMs of membranes in response to anticancer drug induced cell stress promotes expression of p53 mutant proteins and accordant cancer drug resistance.https://doi.org/10.1096/fba.2020-00044colon cancercSrc kinasedrug resistanceglycosphingolipid‐enriched microdomainsglycosphingolipidsmissense mutation |
| spellingShingle | Kartik R. Roy Mohammad B. Uddin Sagor C. Roy Ronald A. Hill John Marshall Yu‐Teh Li Jean Christopher Chamcheu Hua Lu Yong‐Yu Liu Gb3‐cSrc complex in glycosphingolipid‐enriched microdomains contributes to the expression of p53 mutant protein and cancer drug resistance via β‐catenin–activated RNA methylation FASEB BioAdvances colon cancer cSrc kinase drug resistance glycosphingolipid‐enriched microdomains glycosphingolipids missense mutation |
| title | Gb3‐cSrc complex in glycosphingolipid‐enriched microdomains contributes to the expression of p53 mutant protein and cancer drug resistance via β‐catenin–activated RNA methylation |
| title_full | Gb3‐cSrc complex in glycosphingolipid‐enriched microdomains contributes to the expression of p53 mutant protein and cancer drug resistance via β‐catenin–activated RNA methylation |
| title_fullStr | Gb3‐cSrc complex in glycosphingolipid‐enriched microdomains contributes to the expression of p53 mutant protein and cancer drug resistance via β‐catenin–activated RNA methylation |
| title_full_unstemmed | Gb3‐cSrc complex in glycosphingolipid‐enriched microdomains contributes to the expression of p53 mutant protein and cancer drug resistance via β‐catenin–activated RNA methylation |
| title_short | Gb3‐cSrc complex in glycosphingolipid‐enriched microdomains contributes to the expression of p53 mutant protein and cancer drug resistance via β‐catenin–activated RNA methylation |
| title_sort | gb3 csrc complex in glycosphingolipid enriched microdomains contributes to the expression of p53 mutant protein and cancer drug resistance via β catenin activated rna methylation |
| topic | colon cancer cSrc kinase drug resistance glycosphingolipid‐enriched microdomains glycosphingolipids missense mutation |
| url | https://doi.org/10.1096/fba.2020-00044 |
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