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Identification of Novel Protein Expression Changes Following Cisplatin Treatment and Application to Combination
Amy L Stark1,2, Ashraf G Madian1,2, Sawyer W Williams1,2
1Department of Medicine, ‡Committee on Clinical Pharmacology and Pharmacogenomics, ∥Ben May Department for Cancer Research; ⊥Committee on Genetics, Genomics and Systems Biology; #The Institute for Genomics and Systems Biology; ∇Committee on Cancer Biology; and □Department of Human Genetics, The University of Chicago , Chicago, Illinois 60637, United States.
Abstract:
Determining the effect of chemotherapeutic treatment on changes in protein expression can provide important targets for overcoming resistance. Due to challenges in simultaneously measuring large numbers of proteins, a paucity of data exists on global changes. To overcome these challenges, we utilized microwestern arrays that allowed us to measure the abundance and modification state of hundreds of cell signaling and transcription factor proteins in cells following drug exposure. HapMap lymphoblastoid cell lines (LCLs) were exposed to cisplatin, a chemotherapeutic agent commonly used to treat testicular, head and neck, non-small cell lung, and gynecological cancers. We evaluated the expression of 259 proteins following 2, 6, and 12 h of cisplatin treatment in two LCLs with discordant sensitivity to cisplatin. Of these 259 proteins, 66 displayed significantly different protein expression changes (p < 0.05). Fifteen of these proteins were evaluated in a second pair of LCLs with discordant sensitivities to cisplatin; six demonstrated significant differences in expression. We then evaluated a subset of 63 proteins in a second set of LCLs with discordant sensitivity, and 40% of those that were significant in the first pair were also significant in the second part with concordant directionality (p < 0.05). We functionally validated one of the top proteins identified, PDK1, and demonstrated a synergistic relationship between cisplatin and a PDK1 inhibitor in multiple lung cancer lines. This study highlights the potential for identifying novel targets through an understanding of cellular changes in protein expression and modification following drug treatments.
Insights
Chemotherapy resistance can be overcome by identifying new protein targets. This study used microwestern arrays to analyze protein expression changes after cisplatin treatment, revealing potential targets like PDK1 for improved cancer therapies.
Area of Science:
- Proteomics
- Cancer Biology
- Drug Discovery
Background:
- Understanding protein expression changes during chemotherapy is crucial for overcoming drug resistance.
- Measuring global protein alterations after treatment has been limited by technological challenges.
- Microwestern arrays offer a high-throughput method to assess protein abundance and modification states.
Purpose of the Study:
- To investigate global changes in protein expression and modification in response to cisplatin treatment.
- To identify novel protein targets for overcoming chemotherapy resistance.
- To functionally validate identified protein targets in cancer cell lines.
Main Methods:
- Utilized microwestern arrays to measure hundreds of cell signaling and transcription factor proteins.
- Treated HapMap lymphoblastoid cell lines (LCLs) with cisplatin at 2, 6, and 12 hours.
- Evaluated protein expression changes in LCLs with varying sensitivities to cisplatin.
- Functionally validated the role of pyruvate dehydrogenase kinase 1 (PDK1) and its inhibitors.
Main Results:
- Observed significant differential protein expression changes in 66 out of 259 proteins analyzed after cisplatin exposure.
- Identified and validated key proteins, including PDK1, with significant expression differences across multiple LCL pairs.
- Demonstrated a synergistic effect between cisplatin and a PDK1 inhibitor in lung cancer cell lines, suggesting PDK1 as a viable therapeutic target.
Conclusions:
- Microwestern arrays are effective for identifying global protein expression changes induced by chemotherapeutic agents.
- PDK1 is a promising target for combination therapy to enhance cisplatin efficacy in certain cancers.
- This approach aids in discovering novel therapeutic targets by analyzing drug-induced cellular proteomic alterations.
