Protocol for 3D screening of lung cancer spheroids using natural products
Non-small cell lung cancer (NSCLC) is the most common type of lung cancer and accounts for ∼84% of all lung cancer cases. NSCLC remains one of the leading causes of cancer-associated death, with a 5-year survival rate less than 25%. This type of cancer begins with healthy cells that change and start...
| Main Authors: | , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2023-03-01
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| Series: | SLAS Discovery |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2472555223000059 |
| _version_ | 1811158431291146240 |
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| author | Virneliz Fernández Vega Dong Yang Luis Ortiz Jordán Fei Ye Louis Conway Li Yun Chen Justin Shumate Pierre Baillargeon Louis Scampavia Christopher Parker Ben Shen Timothy P. Spicer |
| author_facet | Virneliz Fernández Vega Dong Yang Luis Ortiz Jordán Fei Ye Louis Conway Li Yun Chen Justin Shumate Pierre Baillargeon Louis Scampavia Christopher Parker Ben Shen Timothy P. Spicer |
| author_sort | Virneliz Fernández Vega |
| collection | DOAJ |
| description | Non-small cell lung cancer (NSCLC) is the most common type of lung cancer and accounts for ∼84% of all lung cancer cases. NSCLC remains one of the leading causes of cancer-associated death, with a 5-year survival rate less than 25%. This type of cancer begins with healthy cells that change and start growing out of control, leading to the formation of lesions or tumors. Understanding the dynamics of how the tumor microenvironment promotes cancer initiation and progression that leads to cancer metastasis is crucial to help identify new molecular therapies. 3D primary cell tumor models have received renewed recognition due to their ability to better mimic the complexity of in vivo tumors and as a potential bridge between traditional 2D culture and in vivo studies. Vast improvements in 3D cell culture technologies make them much more cost effective and efficient largely because of the use of a cell-repellent surfaces and a novel angle plate adaptor technology. To exploit this technology, we accessed the Natural Products Library (NPL) at UF Scripps, which consists of crude extracts, partially purified fractions, and pure natural products (NPs). NPs generally are not very well represented in most drug discovery libraries and thus provide new insights to discover leads that could potentially emerge as novel molecular therapies. Herein we describe how we combined these technologies for 3D screening in 1536 well format using a panel of ten NSCLC cells lines (5 wild type and 5 mutant) against ∼1280 selected members of the NPL. After further evaluation, the selected active hits were prioritized to be screened against all 10 NSCLC cell lines as concentration response curves to determine the efficacy and selectivity of the compounds between wild type and mutant 3D cell models. Here, we demonstrate the methods needed for automated 3D screening using microbial NPs, exemplified by crude extracts, partially purified fractions, and pure NPs, that may lead to future use targeting human cancer. |
| first_indexed | 2024-04-10T05:23:23Z |
| format | Article |
| id | doaj.art-3e49c444351c49eaa92e3aaa15039015 |
| institution | Directory Open Access Journal |
| issn | 2472-5552 |
| language | English |
| last_indexed | 2024-04-10T05:23:23Z |
| publishDate | 2023-03-01 |
| publisher | Elsevier |
| record_format | Article |
| series | SLAS Discovery |
| spelling | doaj.art-3e49c444351c49eaa92e3aaa150390152023-03-08T04:14:57ZengElsevierSLAS Discovery2472-55522023-03-012822028Protocol for 3D screening of lung cancer spheroids using natural productsVirneliz Fernández Vega0Dong Yang1Luis Ortiz Jordán2Fei Ye3Louis Conway4Li Yun Chen5Justin Shumate6Pierre Baillargeon7Louis Scampavia8Christopher Parker9Ben Shen10Timothy P. Spicer11Molecular Screening Center, Department of Molecular Medicine, UF Scripps Biomedical Research, Jupiter, FL, USADepartment of Chemistry, UF Scripps Biomedical Research, Jupiter, FL, USA; Natural Products Discovery Center, UF Scripps Biomedical Research, Jupiter, FL, USAMolecular Screening Center, Department of Molecular Medicine, UF Scripps Biomedical Research, Jupiter, FL, USADepartment of Chemistry, UF Scripps Biomedical Research, Jupiter, FL, USADepartment of Chemistry, The Scripps Research Institute, La Jolla, CA, USADepartment of Chemistry, UF Scripps Biomedical Research, Jupiter, FL, USAMolecular Screening Center, Department of Molecular Medicine, UF Scripps Biomedical Research, Jupiter, FL, USAMolecular Screening Center, Department of Molecular Medicine, UF Scripps Biomedical Research, Jupiter, FL, USAMolecular Screening Center, Department of Molecular Medicine, UF Scripps Biomedical Research, Jupiter, FL, USADepartment of Chemistry, The Scripps Research Institute, La Jolla, CA, USADepartment of Chemistry, UF Scripps Biomedical Research, Jupiter, FL, USA; Natural Products Discovery Center, UF Scripps Biomedical Research, Jupiter, FL, USAMolecular Screening Center, Department of Molecular Medicine, UF Scripps Biomedical Research, Jupiter, FL, USA; Corresponding author at: UF Scripps Biomedical Research, 130 Scripps Way #1A1, Jupiter, FL 33458, USA.Non-small cell lung cancer (NSCLC) is the most common type of lung cancer and accounts for ∼84% of all lung cancer cases. NSCLC remains one of the leading causes of cancer-associated death, with a 5-year survival rate less than 25%. This type of cancer begins with healthy cells that change and start growing out of control, leading to the formation of lesions or tumors. Understanding the dynamics of how the tumor microenvironment promotes cancer initiation and progression that leads to cancer metastasis is crucial to help identify new molecular therapies. 3D primary cell tumor models have received renewed recognition due to their ability to better mimic the complexity of in vivo tumors and as a potential bridge between traditional 2D culture and in vivo studies. Vast improvements in 3D cell culture technologies make them much more cost effective and efficient largely because of the use of a cell-repellent surfaces and a novel angle plate adaptor technology. To exploit this technology, we accessed the Natural Products Library (NPL) at UF Scripps, which consists of crude extracts, partially purified fractions, and pure natural products (NPs). NPs generally are not very well represented in most drug discovery libraries and thus provide new insights to discover leads that could potentially emerge as novel molecular therapies. Herein we describe how we combined these technologies for 3D screening in 1536 well format using a panel of ten NSCLC cells lines (5 wild type and 5 mutant) against ∼1280 selected members of the NPL. After further evaluation, the selected active hits were prioritized to be screened against all 10 NSCLC cell lines as concentration response curves to determine the efficacy and selectivity of the compounds between wild type and mutant 3D cell models. Here, we demonstrate the methods needed for automated 3D screening using microbial NPs, exemplified by crude extracts, partially purified fractions, and pure NPs, that may lead to future use targeting human cancer.http://www.sciencedirect.com/science/article/pii/S2472555223000059Non-small lung cancerOrganoidHTSMagnetic bioprintingPhenotypicAngle adaptor |
| spellingShingle | Virneliz Fernández Vega Dong Yang Luis Ortiz Jordán Fei Ye Louis Conway Li Yun Chen Justin Shumate Pierre Baillargeon Louis Scampavia Christopher Parker Ben Shen Timothy P. Spicer Protocol for 3D screening of lung cancer spheroids using natural products SLAS Discovery Non-small lung cancer Organoid HTS Magnetic bioprinting Phenotypic Angle adaptor |
| title | Protocol for 3D screening of lung cancer spheroids using natural products |
| title_full | Protocol for 3D screening of lung cancer spheroids using natural products |
| title_fullStr | Protocol for 3D screening of lung cancer spheroids using natural products |
| title_full_unstemmed | Protocol for 3D screening of lung cancer spheroids using natural products |
| title_short | Protocol for 3D screening of lung cancer spheroids using natural products |
| title_sort | protocol for 3d screening of lung cancer spheroids using natural products |
| topic | Non-small lung cancer Organoid HTS Magnetic bioprinting Phenotypic Angle adaptor |
| url | http://www.sciencedirect.com/science/article/pii/S2472555223000059 |
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