Abstract
Phosphorodiamidate morpholino oligomers (PMO), which have a neutral chemistry, are extensively being used as tools for selective inhibition of gene expression in cell culture models and are currently in human clinical trials. Unlike phosphorothioates (PS ODN) and other charged oligonucleotides, little is known about the uptake characteristics of neutral oligomers. The purpose of this study was to understand the kinetics of PMO transport in cells and correlate with antisense activity. In contrast to primary cells and some transformed cell lines which were uptake permissive, established cancer cell lines showed very poor uptake with an occasional diffuse intracellular pattern. Differential PMO uptake was also observed in immune cells, with dendritic cells and monocytes showing highest uptake compared to T and B cells. In addition, PMO localization was observed to be heterogeneous within a population of uptake permissive cells. Unassisted PMO delivery targeting specific genes was correlated with functional antisense efficacy in experiments showing correction of pre-mRNA missplicing and inhibition of target enzyme activity in cells in culture. PMO internalization in uptakepermissive cells was identified to be specific, saturable, and energy-dependent, suggesting a receptor mediated uptake mechanism. Understanding PMO transport should facilitate the design of more effective synthetic antisense oligomers as therapeutic agents.
Current Pharmaceutical Biotechnology
Title: Cellular Uptake of Neutral Phosphorodiamidate Morpholino Oligomers
Volume: 10 Issue: 6
Author(s): Patrick L. Iversen, Katherine M. Aird, Rebecca Wu, Michael M. Morse and Gayathri R. Devi
Affiliation:
Abstract: Phosphorodiamidate morpholino oligomers (PMO), which have a neutral chemistry, are extensively being used as tools for selective inhibition of gene expression in cell culture models and are currently in human clinical trials. Unlike phosphorothioates (PS ODN) and other charged oligonucleotides, little is known about the uptake characteristics of neutral oligomers. The purpose of this study was to understand the kinetics of PMO transport in cells and correlate with antisense activity. In contrast to primary cells and some transformed cell lines which were uptake permissive, established cancer cell lines showed very poor uptake with an occasional diffuse intracellular pattern. Differential PMO uptake was also observed in immune cells, with dendritic cells and monocytes showing highest uptake compared to T and B cells. In addition, PMO localization was observed to be heterogeneous within a population of uptake permissive cells. Unassisted PMO delivery targeting specific genes was correlated with functional antisense efficacy in experiments showing correction of pre-mRNA missplicing and inhibition of target enzyme activity in cells in culture. PMO internalization in uptakepermissive cells was identified to be specific, saturable, and energy-dependent, suggesting a receptor mediated uptake mechanism. Understanding PMO transport should facilitate the design of more effective synthetic antisense oligomers as therapeutic agents.
Export Options
About this article
Cite this article as:
Iversen L. Patrick, Aird M. Katherine, Wu Rebecca, Morse M. Michael and Devi R. Gayathri, Cellular Uptake of Neutral Phosphorodiamidate Morpholino Oligomers, Current Pharmaceutical Biotechnology 2009; 10 (6) . https://dx.doi.org/10.2174/138920109789069279
DOI https://dx.doi.org/10.2174/138920109789069279 |
Print ISSN 1389-2010 |
Publisher Name Bentham Science Publisher |
Online ISSN 1873-4316 |
- 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
Related Articles
-
Synthesis and In Vitro Evaluation of Some Isatin-Thiazolidinone Hybrid Analogues as Anti-Proliferative Agents
Medicinal Chemistry The Genetic Regulation of ADPRT/PARP-1 in Aging and Cancer Susceptibility
Current Pharmacogenomics Tumor-Derived Extracellular Fragments of Receptor Protein Tyrosine Phosphatases (RPTPs) as Cancer Molecular Diagnostic Tools
Anti-Cancer Agents in Medicinal Chemistry Cannabis-Derived Substances in Cancer Therapy – An Emerging Anti- Inflammatory Role for the Cannabinoids
Current Clinical Pharmacology Editorial (Thematic Issue: Cancer Biomarkers from Bench to Bedside)
Current Cancer Drug Targets The Design of Amphiphilic Polymeric Micelles of Curcumin for Cancer Management
Current Medicinal Chemistry Active Subnetwork GA: A Two Stage Genetic Algorithm Approach to Active Subnetwork Search
Current Bioinformatics Imaging Strategies for Rectal Cancer Initial Staging: Does Pelvic Computed Tomography Provide Significantly Additional Findings when High-resolution Magnetic Resonance Imaging has Been Performed?
Current Medical Imaging Editorial
Current Medicinal Chemistry Factors Associated with Pulmonary Embolism Recurrence and the Benefits of Long-term Anticoagulant Therapy
Cardiovascular & Hematological Disorders-Drug Targets Regulatory T Cells and Cancer Therapy: An Old Story with a New Hope
Current Cancer Therapy Reviews Synthesis and Biological Evaluation of Novel Triazoles Linked 7-hydroxycoumarin as Potent Cytotoxic Agents
Anti-Cancer Agents in Medicinal Chemistry Advances in Exploring the Role of Micrornas in Inflammatory Bowel Disease
MicroRNA Reduction of Oxidative/Nitrosative Stress in Brain and its Involvement in the Neuroprotective Effect of n-3 PUFA in Alzheimer’s Disease
Current Alzheimer Research Sera/Organ Lysates of Selected Animals Living in Polluted Environments Exhibit Cytotoxicity against Cancer Cell Lines
Anti-Cancer Agents in Medicinal Chemistry Tumor Stroma Manipulation By MSC
Current Drug Targets Chemical Synthesis, Characterisation and Biological Evaluation of Furanic- Estradiol Derivatives as Inhibitors of 17β-Hydroxysteroid Dehydrogenase Type 1
Medicinal Chemistry Epidermal Stem Cells - Role in Normal, Wounded and Pathological Psoriatic and Cancer Skin
Current Stem Cell Research & Therapy Release Kinetics from Bio-Polymeric Nanoparticles Encapsulating Protein Synthesis Inhibitor- Cycloheximide, for Possible Therapeutic Applications
Current Pharmaceutical Biotechnology Functions of Ghrelin in Brain, Gut and Liver
CNS & Neurological Disorders - Drug Targets