426 Developing a Molecular Toolkit to define NEK functions in triple-negative breast cancer (TNBC) biology
Objectives/Goals: The never in mitosis kinase (NEK) family regulates vital processes, namely cell cycle progression, but their potential as therapeutic targets in TNBC has not been fully explored. Our studies aim to develop a toolkit to investigate the functional roles of NEKs in pathologies includi...
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| Format: | Article |
| Language: | English |
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Cambridge University Press
2025-04-01
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| Series: | Journal of Clinical and Translational Science |
| Online Access: | https://www.cambridge.org/core/product/identifier/S2059866124010331/type/journal_article |
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| author | Julia Boehling Jack R. Elliot Elizabeth C. Martin Bridgette M. Collins-Burow David H. Drewry Van T. Hoang Sean Lee Matthew E. Burow |
| author_facet | Julia Boehling Jack R. Elliot Elizabeth C. Martin Bridgette M. Collins-Burow David H. Drewry Van T. Hoang Sean Lee Matthew E. Burow |
| author_sort | Julia Boehling |
| collection | DOAJ |
| description | Objectives/Goals: The never in mitosis kinase (NEK) family regulates vital processes, namely cell cycle progression, but their potential as therapeutic targets in TNBC has not been fully explored. Our studies aim to develop a toolkit to investigate the functional roles of NEKs in pathologies including carcinogenesis. Methods/Study Population: To assess differential NEK expression in normal and tumor tissues and correlation of gene expression with patient survival, we used Gene Expression Profiling Interactive Analysis (GEPIA) and Kaplan–Meier Plotter (KMPlot) pan-cancer analysis, respectively. Basal NEK protein levels were determined by immunoblot across a panel of cell lines, including breast cancer, osteosarcoma, hepatocellular carcinoma, and non-cancerous cells, to identify appropriate systems for evaluation of NEK function. Doxycycline-inducible cell lines were generated by transduction with lentiviral stocks of NEK shRNA and overexpression constructs and antibiotic selection. Expression was analyzed by qPCR and immunoblot. Results/Anticipated Results: Expression of NEK2, 4, 5, 6, 8, and 11 was higher in breast tumors compared to normal tissue by GEPIA analysis. Further examination using KMPlot showed a correlation between elevated NEK6 expression and decreased overall survival in patients with aggressive cancers. As an initial proof-of-concept study, we analyzed NEK6 protein expression in breast cancer cells. Levels of NEK6 were elevated in TNBC cells (MDA-MB-231) compared to hormone receptor positive (HR+) breast cancer cells (MCF7). Using complementary approaches to investigate the functional role of NEK6 in breast cancer, we depleted NEK6 expression using shRNAs in TNBC cells and expressed NEK6 in HR+ cells Discussion/Significance of Impact: Because kinase dysregulation promotes oncogenesis and metastasis, targeting kinases is a key strategy in therapeutic development. A NEK-specific molecular toolkit allows researchers to elucidate NEK functions and contributions to carcinogenesis, promoting advancement of novel therapies. |
| format | Article |
| id | doaj-art-a54c8324cf9a4cdcbb50d1b20f0502f7 |
| institution | Kabale University |
| issn | 2059-8661 |
| language | English |
| publishDate | 2025-04-01 |
| publisher | Cambridge University Press |
| record_format | Article |
| series | Journal of Clinical and Translational Science |
| spelling | doaj-art-a54c8324cf9a4cdcbb50d1b20f0502f72025-08-20T03:40:18ZengCambridge University PressJournal of Clinical and Translational Science2059-86612025-04-01912812810.1017/cts.2024.1033426 Developing a Molecular Toolkit to define NEK functions in triple-negative breast cancer (TNBC) biologyJulia Boehling0Jack R. Elliot1Elizabeth C. Martin2Bridgette M. Collins-Burow3David H. Drewry4Van T. Hoang5Sean Lee6Matthew E. Burow7Department of Medicine, Section of Hematology and Oncology, Tulane University, New Orleans, LADepartment of Medicine, Section of Hematology and Oncology, Tulane University, New Orleans, LA Tulane Cancer Center, Tulane Univeristy School of Medicine, New Orleans, LADepartment of Medicine, Section of Hematology and Oncology, Tulane University, New Orleans, LA Tulane Cancer Center, Tulane Univeristy School of Medicine, New Orleans, LADepartment of Medicine, Section of Hematology and Oncology, Tulane University, New Orleans, LA Tulane Cancer Center, Tulane Univeristy School of Medicine, New Orleans, LAUNC Lineberger Comprehensive Cancer Center, School of Medicine, UNC at Chapel Hill, Chapel Hill, NCDepartment of Medicine, Section of Hematology and Oncology, Tulane University, New Orleans, LA Tulane Cancer Center, Tulane Univeristy School of Medicine, New Orleans, LADepartment of Pathology and Laboratory Medicine, Tulane University School of Medicine, New Orleans, LADepartment of Medicine, Section of Hematology and Oncology, Tulane University, New Orleans, LA Tulane Cancer Center, Tulane Univeristy School of Medicine, New Orleans, LAObjectives/Goals: The never in mitosis kinase (NEK) family regulates vital processes, namely cell cycle progression, but their potential as therapeutic targets in TNBC has not been fully explored. Our studies aim to develop a toolkit to investigate the functional roles of NEKs in pathologies including carcinogenesis. Methods/Study Population: To assess differential NEK expression in normal and tumor tissues and correlation of gene expression with patient survival, we used Gene Expression Profiling Interactive Analysis (GEPIA) and Kaplan–Meier Plotter (KMPlot) pan-cancer analysis, respectively. Basal NEK protein levels were determined by immunoblot across a panel of cell lines, including breast cancer, osteosarcoma, hepatocellular carcinoma, and non-cancerous cells, to identify appropriate systems for evaluation of NEK function. Doxycycline-inducible cell lines were generated by transduction with lentiviral stocks of NEK shRNA and overexpression constructs and antibiotic selection. Expression was analyzed by qPCR and immunoblot. Results/Anticipated Results: Expression of NEK2, 4, 5, 6, 8, and 11 was higher in breast tumors compared to normal tissue by GEPIA analysis. Further examination using KMPlot showed a correlation between elevated NEK6 expression and decreased overall survival in patients with aggressive cancers. As an initial proof-of-concept study, we analyzed NEK6 protein expression in breast cancer cells. Levels of NEK6 were elevated in TNBC cells (MDA-MB-231) compared to hormone receptor positive (HR+) breast cancer cells (MCF7). Using complementary approaches to investigate the functional role of NEK6 in breast cancer, we depleted NEK6 expression using shRNAs in TNBC cells and expressed NEK6 in HR+ cells Discussion/Significance of Impact: Because kinase dysregulation promotes oncogenesis and metastasis, targeting kinases is a key strategy in therapeutic development. A NEK-specific molecular toolkit allows researchers to elucidate NEK functions and contributions to carcinogenesis, promoting advancement of novel therapies.https://www.cambridge.org/core/product/identifier/S2059866124010331/type/journal_article |
| spellingShingle | Julia Boehling Jack R. Elliot Elizabeth C. Martin Bridgette M. Collins-Burow David H. Drewry Van T. Hoang Sean Lee Matthew E. Burow 426 Developing a Molecular Toolkit to define NEK functions in triple-negative breast cancer (TNBC) biology Journal of Clinical and Translational Science |
| title | 426 Developing a Molecular Toolkit to define NEK functions in triple-negative breast cancer (TNBC) biology |
| title_full | 426 Developing a Molecular Toolkit to define NEK functions in triple-negative breast cancer (TNBC) biology |
| title_fullStr | 426 Developing a Molecular Toolkit to define NEK functions in triple-negative breast cancer (TNBC) biology |
| title_full_unstemmed | 426 Developing a Molecular Toolkit to define NEK functions in triple-negative breast cancer (TNBC) biology |
| title_short | 426 Developing a Molecular Toolkit to define NEK functions in triple-negative breast cancer (TNBC) biology |
| title_sort | 426 developing a molecular toolkit to define nek functions in triple negative breast cancer tnbc biology |
| url | https://www.cambridge.org/core/product/identifier/S2059866124010331/type/journal_article |
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