13109 Phosphorylation state of Myristoylated Alanine-Rich C-Kinase Substrate Effector Domain mimetics determines its cytotoxicity in glioblastoma and macrophage model
ABSTRACT IMPACT: This study provides insight into how MED2 impacts the immune cells surrounding glioblastoma that help it to grow and spread; having a more complete understanding of how MED2 works will help us better develop therapies that may one day enter the clinic to improve patient outcomes in...
| Main Authors: | , , , , , |
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| Format: | Article |
| Language: | English |
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Cambridge University Press
2021-03-01
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| Series: | Journal of Clinical and Translational Science |
| Online Access: | https://www.cambridge.org/core/product/identifier/S2059866121004039/type/journal_article |
| _version_ | 1811155357929570304 |
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| author | Hasan Alrefai Nicholas J. Eustace Joshua C. Anderson Patricia H. Hicks Peter H. King Christopher D. Willey |
| author_facet | Hasan Alrefai Nicholas J. Eustace Joshua C. Anderson Patricia H. Hicks Peter H. King Christopher D. Willey |
| author_sort | Hasan Alrefai |
| collection | DOAJ |
| description | ABSTRACT IMPACT: This study provides insight into how MED2 impacts the immune cells surrounding glioblastoma that help it to grow and spread; having a more complete understanding of how MED2 works will help us better develop therapies that may one day enter the clinic to improve patient outcomes in glioblastoma. OBJECTIVES/GOALS: The purpose of this study was to determine whether the phosphorylation state of the MED2 peptide impacts its biological activity in GBM and macrophages. MED2 variants include the phosphorylatable wild-type (MED2), pseudo-phosphorylated (MED2-PP), non-phosphorylatable (MED2-NP) and control length (CTL2) peptides. METHODS/STUDY POPULATION: MED2, MED2-NP, MED2-PP, and CTL2 were screened against a panel of molecularly characterized glioblastoma patient derived xenografts and IL4/13 stimulated M2-like THP-1 macrophages. The luminescent cell viability assay, CellTiter-Glo, was used to determine viability. RESULTS/ANTICIPATED RESULTS: The proneural lines XD456 and X1441 were highly sensitive to 5 µM MED2 and 5 µM MED2NP compared to 5 µM MED2PP (p<0.001). There was no statistically significant difference between untreated, 5 µM CTL2, and 5 µM MED2PP groups or between the MED2NP and MED2 treated groups. M2-like THP-1 macrophages were highly sensitive to 10 µM MED2NP compared to 10 µM CTL2 (p<0.01) and 10 µM MED2PP (p<0.01) No statistically significant difference was observed between untreated, 10 µM MED2, 10 µM MED2PP, and 10 µM CTL2 groups. DISCUSSION/SIGNIFICANCE OF FINDINGS: The phosphorylation state of MED2 determines its toxicity. When MED2 is phosphorylated, it is nontoxic to GBM or M2-like macrophages. The non-phosphorylatable version is toxic to both GBM and M2-like macrophages. The wild-type peptide is toxic to GBM but not M2-like macrophages, suggesting that MED2 may be phosphorylated in M2-like macrophages. |
| first_indexed | 2024-04-10T04:31:44Z |
| format | Article |
| id | doaj.art-5804298dd2814a708670a9a997415cd7 |
| institution | Directory Open Access Journal |
| issn | 2059-8661 |
| language | English |
| last_indexed | 2024-04-10T04:31:44Z |
| publishDate | 2021-03-01 |
| publisher | Cambridge University Press |
| record_format | Article |
| series | Journal of Clinical and Translational Science |
| spelling | doaj.art-5804298dd2814a708670a9a997415cd72023-03-10T07:52:03ZengCambridge University PressJournal of Clinical and Translational Science2059-86612021-03-0151110.1017/cts.2021.40313109 Phosphorylation state of Myristoylated Alanine-Rich C-Kinase Substrate Effector Domain mimetics determines its cytotoxicity in glioblastoma and macrophage modelHasan Alrefai0Nicholas J. Eustace1Joshua C. Anderson2Patricia H. Hicks3Peter H. King4Christopher D. Willey5The University of Alabama at Birmingham School of MedicineThe University of Alabama at Birmingham School of MedicineThe University of Alabama at Birmingham School of MedicineThe University of Alabama at Birmingham School of MedicineThe University of Alabama at Birmingham School of MedicineThe University of Alabama at Birmingham School of MedicineABSTRACT IMPACT: This study provides insight into how MED2 impacts the immune cells surrounding glioblastoma that help it to grow and spread; having a more complete understanding of how MED2 works will help us better develop therapies that may one day enter the clinic to improve patient outcomes in glioblastoma. OBJECTIVES/GOALS: The purpose of this study was to determine whether the phosphorylation state of the MED2 peptide impacts its biological activity in GBM and macrophages. MED2 variants include the phosphorylatable wild-type (MED2), pseudo-phosphorylated (MED2-PP), non-phosphorylatable (MED2-NP) and control length (CTL2) peptides. METHODS/STUDY POPULATION: MED2, MED2-NP, MED2-PP, and CTL2 were screened against a panel of molecularly characterized glioblastoma patient derived xenografts and IL4/13 stimulated M2-like THP-1 macrophages. The luminescent cell viability assay, CellTiter-Glo, was used to determine viability. RESULTS/ANTICIPATED RESULTS: The proneural lines XD456 and X1441 were highly sensitive to 5 µM MED2 and 5 µM MED2NP compared to 5 µM MED2PP (p<0.001). There was no statistically significant difference between untreated, 5 µM CTL2, and 5 µM MED2PP groups or between the MED2NP and MED2 treated groups. M2-like THP-1 macrophages were highly sensitive to 10 µM MED2NP compared to 10 µM CTL2 (p<0.01) and 10 µM MED2PP (p<0.01) No statistically significant difference was observed between untreated, 10 µM MED2, 10 µM MED2PP, and 10 µM CTL2 groups. DISCUSSION/SIGNIFICANCE OF FINDINGS: The phosphorylation state of MED2 determines its toxicity. When MED2 is phosphorylated, it is nontoxic to GBM or M2-like macrophages. The non-phosphorylatable version is toxic to both GBM and M2-like macrophages. The wild-type peptide is toxic to GBM but not M2-like macrophages, suggesting that MED2 may be phosphorylated in M2-like macrophages.https://www.cambridge.org/core/product/identifier/S2059866121004039/type/journal_article |
| spellingShingle | Hasan Alrefai Nicholas J. Eustace Joshua C. Anderson Patricia H. Hicks Peter H. King Christopher D. Willey 13109 Phosphorylation state of Myristoylated Alanine-Rich C-Kinase Substrate Effector Domain mimetics determines its cytotoxicity in glioblastoma and macrophage model Journal of Clinical and Translational Science |
| title | 13109 Phosphorylation state of Myristoylated Alanine-Rich C-Kinase Substrate Effector Domain mimetics determines its cytotoxicity in glioblastoma and macrophage model |
| title_full | 13109 Phosphorylation state of Myristoylated Alanine-Rich C-Kinase Substrate Effector Domain mimetics determines its cytotoxicity in glioblastoma and macrophage model |
| title_fullStr | 13109 Phosphorylation state of Myristoylated Alanine-Rich C-Kinase Substrate Effector Domain mimetics determines its cytotoxicity in glioblastoma and macrophage model |
| title_full_unstemmed | 13109 Phosphorylation state of Myristoylated Alanine-Rich C-Kinase Substrate Effector Domain mimetics determines its cytotoxicity in glioblastoma and macrophage model |
| title_short | 13109 Phosphorylation state of Myristoylated Alanine-Rich C-Kinase Substrate Effector Domain mimetics determines its cytotoxicity in glioblastoma and macrophage model |
| title_sort | 13109 phosphorylation state of myristoylated alanine rich c kinase substrate effector domain mimetics determines its cytotoxicity in glioblastoma and macrophage model |
| url | https://www.cambridge.org/core/product/identifier/S2059866121004039/type/journal_article |
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