SLAS Discovery最新文献

Varieties of interactions of anti-CD133 aptamers with cell cultures from patient glioblastoma 抗 CD133 嵌合体与胶质母细胞瘤患者细胞培养物之间的各种相互作用。

IF 2.7 4区生物学

SLAS Discovery Pub Date : 2024-11-17 DOI: 10.1016/j.slasd.2024.100195

Olga Antipova , Valeria Moiseenko , Fatima Dzarieva , Ekaterina Savchenko , Igor Pronin , Galina Pavlova , Alexey Kopylov

{"title":"Varieties of interactions of anti-CD133 aptamers with cell cultures from patient glioblastoma","authors":"Olga Antipova , Valeria Moiseenko , Fatima Dzarieva , Ekaterina Savchenko , Igor Pronin , Galina Pavlova , Alexey Kopylov","doi":"10.1016/j.slasd.2024.100195","DOIUrl":"10.1016/j.slasd.2024.100195","url":null,"abstract":"<div><div>Development of aptatheranostics for glioblastoma (GB) requires investigating aptamer interactions with cells. The paper has described flow cytometry (FC) assessment of direct interactions of fluorescent anti-CD133 aptamers with cells, focusing on cell cultures derived from patient GB (CCPGB). Conventional cell lines with different levels of CD133 mRNA, Caco-2 and HCT116, were used to compare interactions with known 2′FY-RNA aptamer A15 and DNA aptamers of Ap and Cs series, labeled with FAM and Cy5. In addition, interactions of certain non-aptameric oligonucleotides were studied. In the case of antibody interactions with cells, FC signals, mean fluorescence intensities (MFIs), correlated with sizable amounts of CD133 mRNA in Caco-2 cells, and CCPGBs 107 and G01. Unexpectedly, MFI <em>per se</em> could not be the solid indicator of specific interactions of aptamer - CD133/cell. Instead, two types of interactions, target CD133-driven and off-target membrane-associated ones, contribute to MFI. The latter was notably observed for CCPGB Sus/fP2 with tiny CD133 mRNA amount. To prove specificity of aptamer - CD133/cell interactions, titration experiments have been performed, revealing half-saturation concentrations of 120±27 for 2′FY-RNA A15 and 180±12 for DNA Cs5 with Caco-2 cells. This knowledge is an essential step to develop aptatheranostics for GB.</div></div>","PeriodicalId":21764,"journal":{"name":"SLAS Discovery","volume":"29 8","pages":"Article 100195"},"PeriodicalIF":2.7,"publicationDate":"2024-11-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142649938","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

TGF-β receptor-specific NanoBRET Target Engagement in living cells for high-throughput kinase inhibitor screens 活细胞中的 TGF-β 受体特异性 NanoBRET 靶标参与，用于高通量激酶抑制剂筛选。

IF 2.7 4区生物学

SLAS Discovery Pub Date : 2024-11-12 DOI: 10.1016/j.slasd.2024.100196

Marius Wits , Nicole Haarmans , Gonzalo Sanchez-Duffhues , Marie-José Goumans

{"title":"TGF-β receptor-specific NanoBRET Target Engagement in living cells for high-throughput kinase inhibitor screens","authors":"Marius Wits , Nicole Haarmans , Gonzalo Sanchez-Duffhues , Marie-José Goumans","doi":"10.1016/j.slasd.2024.100196","DOIUrl":"10.1016/j.slasd.2024.100196","url":null,"abstract":"<div><div>Targeting transforming growth factor-β (TGF-β) receptors is a promising pharmacological approach to normalize aberrant signaling in genetic and non-genetic TGF-β associated diseases including fibrosis, cancer, cardiovascular and musculoskeletal disorders. To identify novel TGF-β receptor kinase inhibitors, methods like in vitro kinase assays, western blot or transcriptional reporter assays are often used for screening purposes. While these methods may have certain advantages, the lack of integration of key features such as receptor specificity, high-throughput capability, and cellular context resemblance remains a major disadvantage. This deficiency could ultimately hinder the translation of study outcomes into later (clinical) stages of drug development. In this study, we introduce an adjusted and optimized live cell NanoBRET Target Engagement (TE)-based method to identify TGF-β receptor specific kinase inhibitors. This comprehensive toolkit contains various TGF-β type I and type II receptors, with corresponding nanoBRET tracers, and disease-related cell lines, including novel non-commercially available materials. The nanoBRET capacity and kinase inhibitory window can be significantly enhanced for functional measurements when stable expression cell lines and substantially low tracer concentrations are used. In addition, this system can be tailored to study TGF-β associated genetic disorders and possibly be used to screen for disease-specific therapeutics. Therefore, the use of this optimized, live cell, antibody-independent nanoBRET Target Engagement assay is highly encouraged for future high-throughput compound screens targeting TGF-β/BMP receptors.</div></div>","PeriodicalId":21764,"journal":{"name":"SLAS Discovery","volume":"29 8","pages":"Article 100196"},"PeriodicalIF":2.7,"publicationDate":"2024-11-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142634383","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

The history, landscape, and outlook of human cell line authentication and security 人类细胞系鉴定与安全的历史、现状和前景。

IF 2.7 4区生物学

SLAS Discovery Pub Date : 2024-11-08 DOI: 10.1016/j.slasd.2024.100194

Elijah Harbut , Yiorgos Makris , Alexander Pertsemlidis , Leonidas Bleris

引用次数: 0

The development of a novel high-throughput membrane potential assay and a solid-supported membrane (SSM)-based electrophysiological assay to study the pharmacological inhibition of GLUT9/SLC2A9 isoforms in a drug discovery program 开发新型高通量膜电位测定法和基于固体支撑膜（SSM）的电生理学测定法，以研究药物发现计划中对 GLUT9/SLC2A9 同工酶的药理抑制。

IF 2.7 4区生物学

SLAS Discovery Pub Date : 2024-11-08 DOI: 10.1016/j.slasd.2024.100193

Antje Pommereau , Francesca Sassone , Alessandro Poli , Marcella De Silvestris , Lia Scarabottolo , Yasmin Zuschlag , Thomas Licher , Felix Bärenz

引用次数: 0

Development of a live cell assay for real-time monitoring the interactions between the Hippo pathway components 14-3-3 and TAZ 开发实时监测 Hippo 通路成分 14-3-3 和 TAZ 之间相互作用的活细胞检测方法。

IF 2.7 4区生物学

SLAS Discovery Pub Date : 2024-11-05 DOI: 10.1016/j.slasd.2024.100191

Blaž Andlovic , Alexander Wolf , Malgorzata Hiltmann , Bert M. Klebl , Jan Eickhoff , Christian Ottmann

{"title":"Development of a live cell assay for real-time monitoring the interactions between the Hippo pathway components 14-3-3 and TAZ","authors":"Blaž Andlovic , Alexander Wolf , Malgorzata Hiltmann , Bert M. Klebl , Jan Eickhoff , Christian Ottmann","doi":"10.1016/j.slasd.2024.100191","DOIUrl":"10.1016/j.slasd.2024.100191","url":null,"abstract":"<div><div>The Hippo pathway plays an important role in organ size control and tissue homeostasis. Dysregulation is involved in many pathologies, including cancer, which has attracted interest in targeting the Hippo pathway. Since the upstream components are <em>bona fide</em> tumor suppressors, it is feasible to target oncogenic downstream targets such as TAZ, a key downstream effector in the Hippo pathway. Its activity is regulated by phosphorylation on multiple sites, with Ser89 playing a critical role in regulation of TAZ activity. Phosphorylation of TAZ at Ser89 promotes binding to 14–3–3 scaffolding proteins, preventing nuclear translocation and abolishing target gene transcription. Here we describe the development of a cell-based assay suitable for high-throughput screening, based on a split NanoLuc luciferase, for monitoring interactions between 14 3–3 and TAZ in living cells. We have validated the assay by screening of a kinase-biased library. The assay can be quickly adapted for higher throughput and thus offers a valuable tool to study new signal inputs involved in regulation of TAZ activity as well as for identification of molecules that modulate the Hippo pathway.</div></div>","PeriodicalId":21764,"journal":{"name":"SLAS Discovery","volume":"29 8","pages":"Article 100191"},"PeriodicalIF":2.7,"publicationDate":"2024-11-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142607587","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Rapid-response RNA-fluorescence in situ hybridization (FISH) assay platform for coronavirus antiviral high-throughput screening 用于冠状病毒抗病毒高通量筛选的快速反应 RNA 荧光原位杂交 (FISH) 检测平台

IF 2.7 4区生物学

SLAS Discovery Pub Date : 2024-11-04 DOI: 10.1016/j.slasd.2024.100189

Ryan Chan , Christian Shema Mugisha , Vorada Chuenchob , Stephanie A. Moquin , Ujjini H. Manjunatha , Nadine Jarrousse , Vineet D. Menachery , Xuping Xie , Erika L. Flannery , Richard T. Eastman

{"title":"Rapid-response RNA-fluorescence in situ hybridization (FISH) assay platform for coronavirus antiviral high-throughput screening","authors":"Ryan Chan , Christian Shema Mugisha , Vorada Chuenchob , Stephanie A. Moquin , Ujjini H. Manjunatha , Nadine Jarrousse , Vineet D. Menachery , Xuping Xie , Erika L. Flannery , Richard T. Eastman","doi":"10.1016/j.slasd.2024.100189","DOIUrl":"10.1016/j.slasd.2024.100189","url":null,"abstract":"<div><div>Over the past 25 years, the global community has faced challenges posed by three distinct outbreaks of coronaviruses including severe acute respiratory syndrome coronavirus (SARS-CoV), Middle East respiratory syndrome coronavirus (MERS-CoV), and severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). The identification of a novel alphacoronavirus canine CoV (CCoV-HuPn2018) in human patients in Malaysia underscores the potential for crossover infections to humans. The threat of the ever-evolving nature of viral infections as well as the lingering health and socioeconomic effects of the recent SARS-CoV-2 pandemic emphasize the urgent need for advanced antiviral drug screening tools that can be quickly implemented to strengthen preparedness and preventive measures against future outbreaks. Here, we present the development and validation of a novel RNA-fluorescence <em>in situ</em> hybridization (FISH) imaging assay as a 384-well, high-throughput rapid response platform for antiviral drug discovery. RNA-FISH is a powerful tool to visualize specific mRNA in cultured cells using a high-content imaging platform. The flexibility of RNA-FISH probe sets allows for the rapid design of viral genome-specific probes, enabling <em>in vitro</em> assay development to test for inhibition of viral replication by either biologic or small molecule inhibitors. Screening of 170 antiviral compounds in concentration-response demonstrates a strong correlation between the RNA-FISH assay and an immunofluorescence assay (IFA) for both human coronaviruses HCoV-OC43 and HCoV-229E. Additionally, we successfully applied this methodology in the context of CCoV strain 1–71, proving rapid development and deployment, opening new avenues for the evaluation of antiviral drugs to potential future emerging threats.</div></div>","PeriodicalId":21764,"journal":{"name":"SLAS Discovery","volume":"29 8","pages":"Article 100189"},"PeriodicalIF":2.7,"publicationDate":"2024-11-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142578690","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Standards for reporting optical biosensor experiments (STROBE): Improving standards in the reporting of optical biosensor-based data in the literature 光学生物传感器实验报告标准（STROBE）：改进文献中基于光学生物传感器的数据报告标准。

IF 2.7 4区生物学

SLAS Discovery Pub Date : 2024-10-31 DOI: 10.1016/j.slasd.2024.100192

Paul E. Belcher , Anna Moberg , Michael B. Murphy

{"title":"Standards for reporting optical biosensor experiments (STROBE): Improving standards in the reporting of optical biosensor-based data in the literature","authors":"Paul E. Belcher , Anna Moberg , Michael B. Murphy","doi":"10.1016/j.slasd.2024.100192","DOIUrl":"10.1016/j.slasd.2024.100192","url":null,"abstract":"<div><div>The number of peer-reviewed publications that feature biosensor data increases every year. A search of PubMed using common technique terminology, including bio-layer interferometry (BLI), surface plasmon resonance (SPR) and grating-coupled interferometry (GCI) generated more than 2500 scientific papers from 2022. Compared to 2009, when David Myszka and Rebecca Rich presented their most recent review of biosensor literature (Rich and Myszka, 2011), this number has nearly doubled. With this increasing number of publications comes an increasing need for standardization of the way biosensor data is reported in journals to allow for replication of the experiments that were performed. Biosensor data is often poorly described in papers which makes it difficult, if not impossible, to replicate the experiment. Critical information typically missing includes sample preparation, method settings, and data evaluation details. We have also found published work in which the authors have failed to report the type of sensor that was used, or which biosensor instrumentation was used. To come to terms with this growing problem, we propose a standardization of the way biosensor data is reported in scientific journals. We call this standard STROBE, standards for reporting optical biosensor experiments.</div></div>","PeriodicalId":21764,"journal":{"name":"SLAS Discovery","volume":"29 8","pages":"Article 100192"},"PeriodicalIF":2.7,"publicationDate":"2024-10-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142565046","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Magnetic 3D bioprinting of skeletal muscle spheroid for a spheroid-based screening assay 磁性骨骼肌球体三维生物打印，用于基于球体的筛选测定。

IF 2.7 4区生物学

SLAS Discovery Pub Date : 2024-10-28 DOI: 10.1016/j.slasd.2024.100190

Chayanit Chaweewannakorn , Khin The Nu Aye , Joao N. Ferreira

{"title":"Magnetic 3D bioprinting of skeletal muscle spheroid for a spheroid-based screening assay","authors":"Chayanit Chaweewannakorn , Khin The Nu Aye , Joao N. Ferreira","doi":"10.1016/j.slasd.2024.100190","DOIUrl":"10.1016/j.slasd.2024.100190","url":null,"abstract":"<div><div>Over the past decade, there has been a rapid development in the use of magnetic three dimensional (3D) based cell culture systems. Concerning the skeletal muscle, 3D culture systems can provide biological insights for translational clinical research in the fields of muscle physiology and metabolism. These systems can enhance the cell culture environment by improving spatially-oriented cellular assemblies and morphological features closely mimicking the in vivo tissues/organs, since they promote strong interactions between cells and the extracellular matrix (ECM). However, the time-consuming and complex nature of 3D traditional culture techniques pose a challenge to the widespread adoption of 3D systems. Herein, a bench protocol is presented for creating an innovative, promptly assembled and user-friendly culture platform for the magnetic 3D bioprinting of skeletal muscle spheroids. Our protocol findings revealed consistent morphological outcomes and the functional development of skeletal muscle tissue, as evidenced by the expression of muscle-specific contractile proteins and myotubes and the responsiveness to stimulation with cholinergic neurotransmitters. This proof-of-concept protocol confirmed the future potential for further validation and application of spheroid-based assays in human skeletal muscle research.</div></div>","PeriodicalId":21764,"journal":{"name":"SLAS Discovery","volume":"29 8","pages":"Article 100190"},"PeriodicalIF":2.7,"publicationDate":"2024-10-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142570581","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Bioassay protocol metadata annotation: Proposed standards adoption 生物测定协议元数据注释：建议采用的标准。

IF 2.7 4区生物学

SLAS Discovery Pub Date : 2024-10-19 DOI: 10.1016/j.slasd.2024.100188

Rama Balakrishnan , Ellen L. Berg , Christopher C. Butler , Alex M. Clark , Sheryl P. Denker , Isabella Feierberg , Jason Harris , Timothy P. Ikeda , Samantha Jeschonek , Vladimir A. Makarov , Christopher Southan , Dana Vanderwall , Peter Winstanley

引用次数: 0

A mechanism of action-reflective, dual cell-based bioassay for determining the bioactivity of sclerostin-neutralizing antibodies 反映作用机制的双细胞生物测定法，用于确定硬骨蛋白中和抗体的生物活性。

IF 2.7 4区生物学

SLAS Discovery Pub Date : 2024-10-01 DOI: 10.1016/j.slasd.2024.100187

Suzhen Wei , Qiang Wu , Chunlai Cao , Zhuoni Yang , Jianrui Shi , Jingqun Huang , Hua He , Yongjie Lai , Jing Li

{"title":"A mechanism of action-reflective, dual cell-based bioassay for determining the bioactivity of sclerostin-neutralizing antibodies","authors":"Suzhen Wei , Qiang Wu , Chunlai Cao , Zhuoni Yang , Jianrui Shi , Jingqun Huang , Hua He , Yongjie Lai , Jing Li","doi":"10.1016/j.slasd.2024.100187","DOIUrl":"10.1016/j.slasd.2024.100187","url":null,"abstract":"<div><div>Osteoporosis is a major threat to the elderly worldwide. The Wnt signaling pathway plays a critical role in bone development and homeostasis. Sclerostin, a Wnt ligand inhibitor, competes with Wnt ligands for low-density lipoprotein receptor-related protein 5 or 6 (LRP5/6) on osteoblasts, thereby suppressing bone formation. Sclerostin-neutralizing monoclonal antibodies (mAbs) have emerged as a potential bone-forming therapy for osteoporosis. A cell-based bioassay which determines the relative activity of a product, related to its mechanism of action, is of great importance from drug discovery to quality control and batch release. Currently used cell-based bioassays for sclerostin-neutralizing mAbs usually use Wnt1 or Wnt3a to stimulate the Wnt pathway; sclerostin is a direct inhibitor of Wnt1 but not Wnt3a. Wnt1 is a highly hydrophobic protein that binds to the producing cell membrane and acts in a juxtacrine manner to stimulate the Wnt pathway in neighboring cells. Bioassays for drugs that induce Wnt1 signaling should be performed in a juxtacrine manner. Here, we present a mechanism of action-reflective, dual cell-based reporter gene assay. In this assay, Wnt1 producer cells are co-cultured with cells containing the Wnt reporter genes, Wnt1 on the producer cells activates the Wnt signaling pathway in the reporter cells that are in direct cell-to-cell contact, and sclerostin-neutralizing mAbs specifically and effectively antagonize the sclerostin-mediated Wnt reporter gene suppression. This bioassay demonstrates good specificity, accuracy, linearity, and precision and is suitable for quality control, stability testing, batch release, and biosimilarity assessment of sclerostin-neutralizing mAbs.</div></div>","PeriodicalId":21764,"journal":{"name":"SLAS Discovery","volume":"29 7","pages":"Article 100187"},"PeriodicalIF":2.7,"publicationDate":"2024-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142402302","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0