SLAS Discovery: Advancing the Science of Drug Discovery最新文献

In Vitro Three-dimensional (3D) Cell Culture Tools for Spheroid and Organoid Models 用于球形和类器官模型的体外三维（3D）细胞培养工具

SLAS Discovery: Advancing the Science of Drug Discovery Pub Date : 2023-12-13 DOI: 10.1016/j.slasd.2023.12.003

Sang-Yun Lee, In-Seong Koo, Hyun Ju Hwang, Dong Woo Lee

{"title":"In Vitro Three-dimensional (3D) Cell Culture Tools for Spheroid and Organoid Models","authors":"Sang-Yun Lee, In-Seong Koo, Hyun Ju Hwang, Dong Woo Lee","doi":"10.1016/j.slasd.2023.12.003","DOIUrl":"https://doi.org/10.1016/j.slasd.2023.12.003","url":null,"abstract":"Three-dimensional (3D) cell culture technology has been steadily studied since the 1990’s due to its superior biocompatibility compared to the conventional two-dimensional (2D) cell culture technology, and has recently developed into an organoid culture technology that further improved biocompatibility. Since the 3D culture of human cell lines in artificial scaffolds was demonstrated in the early 90′s, 3D cell culture technology has been actively developed owing to various needs in the areas of disease research, precision medicine, new drug development, and some of these technologies have been commercialized. In particular, 3D cell culture technology is actively being applied and utilized in drug development and cancer-related precision medicine research. Drug development is a long and expensive process that involves multiple steps—from target identification to lead discovery and optimization, preclinical studies, and clinical trials for approval for clinical use. Cancer ranks first among life-threatening diseases owing to intra-tumoral heterogeneity associated with metastasis, recurrence, and treatment resistance, ultimately contributing to treatment failure and adverse prognoses. Therefore, there is an urgent need for the development of efficient drugs using 3D cell culture techniques that can closely mimic in vivo cellular environments and customized tumor models that faithfully represent the tumor heterogeneity of individual patients. This review discusses 3D cell culture technology focusing on research trends, commercialization status, and expected effects developed until recently. We aim to summarize the great potential of 3D cell culture technology and contribute to expanding the base of this technology.","PeriodicalId":501832,"journal":{"name":"SLAS Discovery: Advancing the Science of Drug Discovery","volume":"46 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2023-12-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"138693360","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Reduced levels of serum EPA and DHA identified in patients with non-small-cell lung cancer using a new rapid validated LC-MS/MS method 利用新型快速验证 LC-MS/MS 方法确定非小细胞肺癌患者血清 EPA 和 DHA 水平降低的原因

SLAS Discovery: Advancing the Science of Drug Discovery Pub Date : 2023-12-13 DOI: 10.1016/j.slasd.2023.12.006

Yi Wang, Tongxin Yin, Jiaoyuan Li, Xia Luo, Ke Liu, Tingting Long, Ying Shen, Liming Cheng

{"title":"Reduced levels of serum EPA and DHA identified in patients with non-small-cell lung cancer using a new rapid validated LC-MS/MS method","authors":"Yi Wang, Tongxin Yin, Jiaoyuan Li, Xia Luo, Ke Liu, Tingting Long, Ying Shen, Liming Cheng","doi":"10.1016/j.slasd.2023.12.006","DOIUrl":"https://doi.org/10.1016/j.slasd.2023.12.006","url":null,"abstract":"Background: Eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) have been suggested to play roles in various diseases, yet there is little data on their changes in patients with non-small-cell lung cancer (NSCLC). A simple LC-MS/MS method for EPA and DHA determination is critical to exploring EPA and DHA level changes in NSCLC patients.Method: 25 μL of serum was mixed with 25 μL of internal standard working solution, and then 450 μL of acetonitrile for protein precipitation. After vortex and centrifugation, the supernatant was directly used for LC-MS/MS analysis. The method was well validated with linearity, precision, recovery, and matrix effect. The concentrations of EPA and DHA in serum samples from 211 NSCLC patients and 227 healthy controls were determined by this LC-MS/MS method.Results: Good separation and reliable quantification of EPA and DHA in serum samples were achieved by our method. Compared with healthy controls, serum EPA and DHA were significantly reduced in both adenocarcinoma and squamous cell carcinoma. The concentrations of EPA and DHA showed a progressive decrease in healthy controls, early- and advanced-stage NSCLC patients.Conclusions: This study identified significant reductions in serum EPA and DHA in NSCLC patients through the development of an LC-MS/MS method.","PeriodicalId":501832,"journal":{"name":"SLAS Discovery: Advancing the Science of Drug Discovery","volume":"12 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2023-12-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"138693359","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Patient derived glioma stem cell spheroid reporter assays for live cell high content analysis 用于活细胞高含量分析的患者胶质瘤干细胞球形报告实验

SLAS Discovery: Advancing the Science of Drug Discovery Pub Date : 2023-12-13 DOI: 10.1016/j.slasd.2023.12.005

Jayne Culley, Peter W Nagle, John C Dawson, Neil O Carragher

引用次数: 0

Protocol for 3D Drug Sensitivity and Resistance Testing of Patient-Derived Cancer Cells in 384-Well Plates 在 384 孔板中对患者衍生癌症细胞进行三维药物敏感性和耐药性测试的方案

SLAS Discovery: Advancing the Science of Drug Discovery Pub Date : 2023-12-13 DOI: 10.1016/j.slasd.2023.12.004

Michaela Feodoroff, Piia Mikkonen, Mariliina Arjama, Astrid Murumägi, Olli Kallioniemi, Swapnil Potdar, Laura Turunen, Vilja Pietiäinen

{"title":"Protocol for 3D Drug Sensitivity and Resistance Testing of Patient-Derived Cancer Cells in 384-Well Plates","authors":"Michaela Feodoroff, Piia Mikkonen, Mariliina Arjama, Astrid Murumägi, Olli Kallioniemi, Swapnil Potdar, Laura Turunen, Vilja Pietiäinen","doi":"10.1016/j.slasd.2023.12.004","DOIUrl":"https://doi.org/10.1016/j.slasd.2023.12.004","url":null,"abstract":"Establishment of drug testing of patient-derived cancer cells (PDCs) in physiologically relevant 3-dimensional (3D) culture is central for drug discovery and cancer research, as well as for functional precision medicine. Here, we describe the detailed protocol allowing the 3D drug testing of PDCs – or any type of cells of interest – in Matrigel in 384-well plate format using automation. We also provide an alternative protocol, which does not require supporting matrices. The cancer tissue is obtained directly from clinics (after surgery or biopsy) and processed into single cell suspension. Systematic drug sensitivity and resistance testing (DSRT) is carried out on the PDCs directly after cancer cell isolation from tissue or on cells expanded for a few passages. In the 3D-DSRT assay, the PDCs are plated in 384-well plates in Matrigel, grown as spheroids, and treated with compounds of interest for 72 h. The cell viability is directly measured using a luminescence-based assay. Alternatively, prior to the cell viability measurement, drug-treated cells can be directly subjected to automated high-content bright field imaging or stained for fluorescence (live) cell microscopy for further image analysis. This is followed by the quality control and data analysis. The 3D-DSRT can be performed within a 1–3-week timeframe of the clinical sampling of cancer tissue, depending on the amount of the obtained tissue, growth rate of cancer cells, and the number of drugs being tested. The 3D-DSRT method can be flexibly modified, e.g., to be carried out with or without supporting matrices with U-bottom 384-well plates when appropriate for the PDCs or other cell models used.","PeriodicalId":501832,"journal":{"name":"SLAS Discovery: Advancing the Science of Drug Discovery","volume":"1 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2023-12-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"138693357","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Reprint of: Detection and Impact of Hypoxic Regions in Multicellular Tumor Spheroid Cultures formed by Head and Neck Squamous Cell Carcinoma Cells Lines 重印：头颈部鳞状细胞癌细胞系形成的多细胞肿瘤球状培养物中缺氧区的检测及其影响

SLAS Discovery: Advancing the Science of Drug Discovery Pub Date : 2023-12-13 DOI: 10.1016/j.slasd.2023.12.002

David A. Close, Paul A. Johnston

{"title":"Reprint of: Detection and Impact of Hypoxic Regions in Multicellular Tumor Spheroid Cultures formed by Head and Neck Squamous Cell Carcinoma Cells Lines","authors":"David A. Close, Paul A. Johnston","doi":"10.1016/j.slasd.2023.12.002","DOIUrl":"https://doi.org/10.1016/j.slasd.2023.12.002","url":null,"abstract":"In solid tumors like head and neck cancer (HNC), chronic and acute hypoxia have serious adverse clinical consequences including poorer overall patient prognosis, enhanced metastasis, increased genomic instability, and resistance to radiation-, chemo-, or immuno-therapies. However, cells in the two-dimensional monolayer cultures typically used for cancer drug discovery experience 20%-21% O2 levels (normoxic) which are 4-fold higher than O2 levels in normal tissues and ≥10-fold higher than in the hypoxic regions of solid tumors. The oxygen electrodes, exogenous bio-reductive markers, and increased expression of endogenous hypoxia-regulated proteins like HIF-1α generally used to mark hypoxic regions in solid tumors are impractical in large sample numbers and longitudinal studies. We used a novel homogeneous live-cell permeant HypoxiTRAK™ (HPTK) molecular probe compatible with high content imaging detection, analysis, and throughput to identify and quantify hypoxia levels in live HNC multicellular tumor spheroid (MCTS) cultures over time. Accumulation of fluorescence HPTK metabolite in live normoxic HNC MCTS cultures correlated with hypoxia detection by both pimonidazole and HIF-1α staining. In HNC MCTSs, hypoxic cytotoxicity ratios for the hypoxia activated prodrugs (HAP) evofosfamide and tirapazamine were much smaller than have been reported for uniformly hypoxic 2D monolayers in gas chambers, and many viable cells remained after HAP exposure. Cells in solid tumors and MCTSs experience three distinct O2 microenvironments dictated by their distances from blood vessels or MCTS surfaces, respectively; oxic, hypoxic, or intermediate levels of hypoxia. These studies support the application of more physiologically relevant in vitro 3D models that recapitulate the heterogeneous microenvironments of solid tumors for preclinical cancer drug discovery.","PeriodicalId":501832,"journal":{"name":"SLAS Discovery: Advancing the Science of Drug Discovery","volume":"1 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2023-12-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"138693361","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0