Abstract
Neural differentiation of embryonic stem cells (ESC) is considered a promising model to perform in vitro testing for neuroactive and neurotoxic compounds. We studied the potential of a dual reporter murine ESC line to identify bioactive and/or toxic compounds. This line expressed firefly luciferase under the control of the neural cell-specific tubulin alpha promoter (TUBA1A), and renilla luciferase under the control of the ubiquitous translation elongation factor 1-alpha-1 (EEF1A1) promoter. During neural differentiation, TUBA1A activity increased, while EEF1A1 activity decreased. We first validated our test system using the known neurotoxin methyl mercury. This compound altered expression of both reporter genes, with ESC-derived neural precursors being affected at markedly lower concentrations than undifferentiated ESCs. Analysis of a library of 1040 bioactive compounds picked up 127 compounds with altered EEF1A1 and/or TUBA1A promoter activity, which were classified in 4 clusters. Cluster 1 (low EEF1A1 and TUBA1A) was the largest cluster, containing many cytostatic drugs, as well as known neurodevelopmental toxicants, psychotropic drugs and endocrine disruptors. Cluster 2 (high EEF1A1, stable TUBA1A) was limited to three sulfonamides. Cluster 3 (high EEF1A1 and TUBA1A) was small, but markedly enriched in neuroactive and neurotoxic compounds. Cluster 4 (stable EEF1A1, high TUBA1A) was heterogeneous, containing endocrine disruptors, neurotoxic and cytostatic drugs. The dual reporter gene assay described here might be a useful addition to in vitro drug testing panels. Our two-dimensional testing strategy provides information on complex response patterns, which could not be achieved by a single marker approach.
Keywords: Developmental toxicity, embryonic stem cells, ESC, in vitro screening, neural differentiation, neurotoxicology, Cluster Analysis, Neural Cells, neurotoxic compounds, alpha promoter.
Current Medicinal Chemistry
Title:Embryonic Stem Cell-Based Screen for Small Molecules: Cluster Analysis Reveals Four Response Patterns in Developing Neural Cells
Volume: 20 Issue: 5
Author(s): I. Kern, R. Xu, S. Julien, D. M. Suter, O. Preynat-Seauve, M. Baquie, A. Poncet, C. Combescure, L. Stoppini, C. V. Thriel and Karl-Heinz Krause
Affiliation:
Keywords: Developmental toxicity, embryonic stem cells, ESC, in vitro screening, neural differentiation, neurotoxicology, Cluster Analysis, Neural Cells, neurotoxic compounds, alpha promoter.
Abstract: Neural differentiation of embryonic stem cells (ESC) is considered a promising model to perform in vitro testing for neuroactive and neurotoxic compounds. We studied the potential of a dual reporter murine ESC line to identify bioactive and/or toxic compounds. This line expressed firefly luciferase under the control of the neural cell-specific tubulin alpha promoter (TUBA1A), and renilla luciferase under the control of the ubiquitous translation elongation factor 1-alpha-1 (EEF1A1) promoter. During neural differentiation, TUBA1A activity increased, while EEF1A1 activity decreased. We first validated our test system using the known neurotoxin methyl mercury. This compound altered expression of both reporter genes, with ESC-derived neural precursors being affected at markedly lower concentrations than undifferentiated ESCs. Analysis of a library of 1040 bioactive compounds picked up 127 compounds with altered EEF1A1 and/or TUBA1A promoter activity, which were classified in 4 clusters. Cluster 1 (low EEF1A1 and TUBA1A) was the largest cluster, containing many cytostatic drugs, as well as known neurodevelopmental toxicants, psychotropic drugs and endocrine disruptors. Cluster 2 (high EEF1A1, stable TUBA1A) was limited to three sulfonamides. Cluster 3 (high EEF1A1 and TUBA1A) was small, but markedly enriched in neuroactive and neurotoxic compounds. Cluster 4 (stable EEF1A1, high TUBA1A) was heterogeneous, containing endocrine disruptors, neurotoxic and cytostatic drugs. The dual reporter gene assay described here might be a useful addition to in vitro drug testing panels. Our two-dimensional testing strategy provides information on complex response patterns, which could not be achieved by a single marker approach.
Export Options
About this article
Cite this article as:
Kern I., Xu R., Julien S., Suter M. D., Preynat-Seauve O., Baquie M., Poncet A., Combescure C., Stoppini L., Thriel V. C. and Krause Karl-Heinz, Embryonic Stem Cell-Based Screen for Small Molecules: Cluster Analysis Reveals Four Response Patterns in Developing Neural Cells, Current Medicinal Chemistry 2013; 20 (5) . https://dx.doi.org/10.2174/092986713804999330
DOI https://dx.doi.org/10.2174/092986713804999330 |
Print ISSN 0929-8673 |
Publisher Name Bentham Science Publisher |
Online ISSN 1875-533X |
Call for Papers in Thematic Issues
Advances in Medicinal Chemistry: From Cancer to Chronic Diseases.
The broad spectrum of the issue will provide a comprehensive overview of emerging trends, novel therapeutic interventions, and translational insights that impact modern medicine. The primary focus will be diseases of global concern, including cancer, chronic pain, metabolic disorders, and autoimmune conditions, providing a broad overview of the advancements in ...read more
Approaches to the treatment of chronic inflammation
Chronic inflammation is a hallmark of numerous diseases, significantly impacting global health. Although chronic inflammation is a hot topic, not much has been written about approaches to its treatment. This thematic issue aims to showcase the latest advancements in chronic inflammation treatment and foster discussion on future directions in this ...read more
Cellular and Molecular Mechanisms of Non-Infectious Inflammatory Diseases: Focus on Clinical Implications
The Special Issue covers the results of the studies on cellular and molecular mechanisms of non-infectious inflammatory diseases, in particular, autoimmune rheumatic diseases, atherosclerotic cardiovascular disease and other age-related disorders such as type II diabetes, cancer, neurodegenerative disorders, etc. Review and research articles as well as methodology papers that summarize ...read more
Chalcogen-modified nucleic acid analogues
Chalcogen-modified nucleosides, nucleotides and oligonucleotides have been of great interest to scientific research for many years. The replacement of oxygen in the nucleobase, sugar or phosphate backbone by chalcogen atoms (sulfur, selenium, tellurium) gives these biomolecules unique properties resulting from their altered physical and chemical properties. The continuing interest in ...read more
- Author Guidelines
- Graphical Abstracts
- Fabricating and Stating False Information
- Research Misconduct
- Post Publication Discussions and Corrections
- Publishing Ethics and Rectitude
- Increase Visibility of Your Article
- Archiving Policies
- Peer Review Workflow
- Order Your Article Before Print
- Promote Your Article
- Manuscript Transfer Facility
- Editorial Policies
- Allegations from Whistleblowers
- Announcements
Related Articles
-
Current Status of Carbapenem Antibiotics
Current Topics in Medicinal Chemistry Perspectives in Genomics The Future of Fungi in ‘omics’ era
Current Genomics Geriatric Evaluation of Oncological Elderly Patients
Anti-Cancer Agents in Medicinal Chemistry Recent Advances in Peptide Nucleic Acids as Antibacterial Agents
Current Medicinal Chemistry A Discussion of Natural Rubber Latex Allergy with Special Reference to Children: Clinical Considerations
Current Drug Targets - Immune, Endocrine & Metabolic Disorders Itraconazole vs Fluconazole as a Primary Prophylaxis for Fungal Infections in HIV-Infected Patients in Thailand
Current HIV Research Nociceptive Pathway and Pathology of Low Back Pain
Current Rheumatology Reviews Saving the Ischemic Penumbra: Potential Role for Statins and Phosphodiesterase Inhibitors
Current Vascular Pharmacology Immunosuppression in Liver Transplantation
Current Drug Targets Current Status of Anti-Picornavirus Therapies
Current Pharmaceutical Design Molecular Targets of Tannic Acid in Alzheimer's Disease
Current Alzheimer Research Nano-Neurotherapeutics (NNTs): An Emergent and Multifaceted Tool for CNS Disorders
Current Drug Metabolism Novelty in Inflammation and Immunomodulation in Migraine
Current Pharmaceutical Design Multimodalities Imaging of Immunoglobulin 4-related Cardiovascular Disorders
Current Cardiology Reviews The Role of Macrophage Migration Inhibitory Factor in Critical Illness
Mini-Reviews in Medicinal Chemistry Sepsis and Solid Organ Transplantation
Current Drug Targets Community-Acquired Pneumonia in Children
Recent Patents on Inflammation & Allergy Drug Discovery Rediscovering the Antibiotics of the Hive
Recent Patents on Anti-Infective Drug Discovery Biologic Therapy in Psoriasis: Safety Profile
Current Drug Safety Rationale of Hyperbaric Oxygenation in Cerebral Vascular Insult
Current Vascular Pharmacology