The goal of this study was to identify novel drug repurposing opportunities in cancer by utilizing the off-target profiles of clinically relevant kinase inhibitors. This was based on the observation that the global target profiles of compounds are largely ignored and that many compounds have activity that cannot be explained by their cognate target alone. Additionally, by utilizing clinically relevant compounds, any results would hold a high potential for eventual clinical implementation. We utilized a systems pharmacology approach utilizing cell viability-based drug screening to identify compounds with beneficial off-target activity and then using chemical and phosphoproteomics in order to elucidate the mechanisms of action of these compounds. We found that tivantinib has off-target activity in NSCLC cells through inhibition of GSK3. Based on tivantinib’s ability to inhibit GSK3, we hypothesized that tivantinib would therefore have activity in acute myeloid leukemia (AML). We found that tivantinib had potent activity in AML through inhibition of GSK3. We also identified a highly synergistic combination with ABT-199 by drug synergy screening which was effective in HL60 cells and patient derived AML cells. We also found that the anaplastic lymphoma kinase (ALK) inhibitor, ceritinib, had activity across several ALK-negative lung cancer cell lines. We utilized integrated functional proteomics to identify the new targets and network-wide signaling effects. Combining pharmacological inhibitors and RNA interference revealed a polypharmacology mechanism involving the noncanonical targets IGF1R, FAK1, RSK1 and RSK2. Mutating the downstream signaling hub YB1 protected cells from ceritinib. Consistent with YB1 signaling being known to cause taxol resistance, combination of ceritinib with paclitaxel displayed strong synergy, particularly in cells expressing high FAK autophosphorylation, which we show to be prevalent in lung cancer. Together, we present a systems chemical biology platform for elucidating multikinase inhibitor mechanisms, synergistic drug combinations, mechanistic biomarker candidates and identifying novel drug repurposing opportunities.
| Identifer | oai:union.ndltd.org:USF/oai:scholarcommons.usf.edu:etd-8886 |
| Date | 30 May 2018 |
| Creators | Kuenzi, Brent M. |
| Publisher | Scholar Commons |
| Source Sets | University of South Flordia |
| Detected Language | English |
| Type | text |
| Format | application/pdf |
| Source | Graduate Theses and Dissertations |
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