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Extracellular Protein Microarray Technology for High Throughput Detection of Low Affinity Receptor-Ligand Interactions
Published on: January 7, 2019
Reverse phase protein microarrays and their utility in drug development
Maria Isabella Sereni1, Mariaelena Pierobon, Roberto Angioli
1Center for Applied Proteomics and Molecular Medicine, George Mason University, Manassas, VA, USA.
Abstract:
The majority of human diseases, including cancer, are characterized by abnormal protein function. Proteins regulate virtually every cellular process and exhibit multiple kinds of post-translational modification that modulate expression levels and activation states, such as phosphorylation by protein kinases. Additionally proteins interact with each other in complex regulatory networks and signal transduction pathways modulated by feedback mechanisms. These pathways are disrupted in disease and altered by therapeutic drugs. Reverse phase protein microarray (RPMA) technology allows simultaneous measurement of numerous phosphorylated, glycosylated, cleaved, or total cellular proteins from complex mixtures in many samples at once. Therefore, RPMAs can provide a portrait of a cell's signaling pathways in diseased states, before and after treatment with drugs, and allows comparison of changes in drug-resistant and sensitive cells. Furthermore, the technology offers a means of connecting genomic abnormalities in cancer to targetable alterations in protein signaling pathways, even for genetic events that seem otherwise undruggable. Consequently, the RPMA platform has great utility in many steps of drug development including target identification, validation of a pharmaceutical agent's efficacy, understanding mechanisms of action, and discovery of biomarkers that predict or guide therapeutic response. RPMAs have become a powerful tool for drug development and are now being integrated into human clinical cancer trials, where they are being used to personalize therapy.
Insights
Reverse phase protein microarrays (RPMAs) enable simultaneous measurement of cellular proteins, offering insights into disease signaling pathways. This technology aids drug development by identifying targets, validating efficacy, and personalizing cancer therapy.
Area of Science:
- Biochemistry
- Molecular Biology
- Cancer Research
Background:
- Human diseases, including cancer, often stem from abnormal protein function.
- Cellular processes are regulated by proteins, their post-translational modifications (e.g., phosphorylation), and complex signaling networks.
- Disruptions in these pathways are implicated in disease and affected by therapeutic interventions.
Purpose of the Study:
- To highlight the utility of Reverse Phase Protein Microarray (RPMA) technology in understanding cellular signaling.
- To demonstrate RPMA's role in disease research, drug development, and personalized medicine.
Main Methods:
- Utilizes Reverse Phase Protein Microarray (RPMA) technology for high-throughput protein analysis.
- Enables simultaneous measurement of various protein states (phosphorylated, total, etc.) across multiple samples.
- Facilitates comparison of signaling pathway alterations in diseased vs. healthy states, and drug-resistant vs. sensitive cells.
Main Results:
- RPMA provides a comprehensive view of cellular signaling pathways in disease contexts.
- The technology connects genomic alterations in cancer to actionable protein signaling changes.
- RPMA aids in identifying drug targets, validating drug efficacy, and understanding mechanisms of action.
Conclusions:
- RPMA is a powerful tool for drug discovery and development, from target identification to biomarker discovery.
- RPMA facilitates the understanding of drug resistance mechanisms and the personalization of cancer therapy.
- The integration of RPMA into clinical trials signifies its growing importance in precision oncology.
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