Taxane-Platin-Resistant Lung Cancers Co-develop Hypersensitivity to JumonjiC Demethylase Inhibitors.
ABSTRACT: Although non-small cell lung cancer (NSCLC) patients benefit from standard taxane-platin chemotherapy, many relapse, developing drug resistance. We established preclinical taxane-platin-chemoresistance models and identified a 35-gene resistance signature, which was associated with poor recurrence-free survival in neoadjuvant-treated NSCLC patients and included upregulation of the JumonjiC lysine demethylase KDM3B. In fact, multi-drug-resistant cells progressively increased the expression of many JumonjiC demethylases, had altered histone methylation, and, importantly, showed hypersensitivity to JumonjiC inhibitors in vitro and in vivo. Increasing taxane-platin resistance in progressive cell line series was accompanied by progressive sensitization to JIB-04 and GSK-J4. These JumonjiC inhibitors partly reversed deregulated transcriptional programs, prevented the emergence of drug-tolerant colonies from chemo-naive cells, and synergized with standard chemotherapy in vitro and in vivo. Our findings reveal JumonjiC inhibitors as promising therapies for targeting taxane-platin-chemoresistant NSCLCs.
Project description:While taxane-platin standard chemotherapy provides benefit in advanced and localized non-small cell lung cancer (NSCLC), the majority of patients relapse with drug resistant tumors. Mechanisms underlying NSCLC resistance to this standard doublet chemotherapy are still not fully understood, and treatment options for chemoresistant lung tumors are limited. The goals of this work were to establish new preclinical NSCLC models of resistance to taxane-platin doublet chemotherapy, identify mechanisms of resistance, and develop new rational pharmacologic approaches to target drug resistant NSCLCs. Overall design: Isogenic drug resistant cell lines were established by long-term treatment of chemosensitive, parental NSCLC cell lines (NCI-H1299 and NCI-H1355) with drug on/drug off cycles of paclitaxel + carboplatin doublet chemotherapy given in a clinically relevant 2:3 paclitaxel-to-carboplatin ratio. Total RNA was extracted and microarrays were performed on parental cell lines (used as control) and all developed resistant cell line variants (denoted as T followed by number of chemotherapy cycles). Samples included biological replicates (5 for parental, 2 for each intermediate resistance variant, 3 for the most resistant variant). Subcutaneous xenografts of H1299 parental cell line and the resistant H1299 T18 variant were also profiled by microarrays (3 tumors per group). 35-gene resistance signature was derived from cell lines and xenografts, and evaluated in neoadjuvant treated NSCLC patients to identify a clinically relevant, druggable target. Pharmacological targeting using JumonjiC histone lysine demethylase inhibitors, JIB-04 and GSK-J4, resulted in selective killing of chemoresistant cells over parental cell lines. To reveal mechanistic insights into JumonjiC inhibitor sensitization, H1299 Parental and H1299 T18 resistant cells were each treated with either DMSO, 0.2 µM JIB-04 or 1 µM GSK-J4 for 24 h, and studied by microarrays (2 biological replicates per treatment).
Project description:Characterization of histone 3 lysine 4 and lysine 27 trimethylation changes upon development of taxane-platin drug resistance in NSCLC cells and evaluation of these histone modifications after treatment with Jumonji KDM inhibitors, JIB-04 and GSK-J4. Overall design: Comparison of H3K4me3 and H3K27me3 profiles between H1299 Parental cells (chemo-sensitive) and H1299 T18 cells (taxane-platin resistant), and assessment of Jumonji KDM inhibitors on H1299 T18 cells (JIB-04 treated vs DMSO treated, and GSK-J4 treated vs. DMSO treated).
Project description:Characterization of gene expression changes upon development of taxane-platin drug resistance in NSCLC cells and further, upon treatment of these resistant cells with the Jumonji KDM inhibitor, GSK-J4. Overall design: Comparison of gene expression changes between H1299 Parental cells (chemo-sensitive) and H1299 T18 cells (taxane-platin resistant), and comparison of H1299 T18: GSK-J4 treated vs. H1299 T18: DMSO control.
Project description:BACKGROUND: Ovarian cancer is the leading cause of death from gynecologic cancer in women worldwide. According to the National Cancer Institute, ovarian cancer has the highest mortality rate among all the reproductive cancers in women. Advanced stage diagnosis and chemo/radio-resistance is a major obstacle in treating advanced ovarian cancer. The most commonly employed chemotherapeutic drug for ovarian cancer treatment is cis-platin. As with most chemotherapeutic drugs, many patients eventually become resistant to cis-platin and therefore, diminishing its effect. The efficacy of current treatments may be improved by increasing the sensitivity of cancer cells to chemo/radiation therapies. METHODS: The present study is focused on identifying the differential expression of regulatory microRNAs (miRNAs) between cis-platin sensitive (A2780), and cis-platin resistant (A2780/CP70) cell lines. Cell proliferation assays were conducted to test the sensitivity of the two cell lines to cis-platin. Differential expression patterns of miRNA between cis-platin sensitive and cis-platin resistant cell lines were analyzed using novel LNA technology. RESULTS: Our results revealed changes in expression of 11 miRNAs out of 1,500 miRNAs analyzed. Out of the 11 miRNAs identified, 5 were up-regulated in the A2780/CP70 cell line and 6 were down regulated as compared to cis-platin sensitive A2780 cells. Our microRNA data was further validated by quantitative real-time PCR for these selected miRNAs. Ingenuity Pathway Analysis (IPA) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis was performed for the selected miRNAs and their putative targets to identify the potential pathways and networks involved in cis-platin resistance. CONCLUSIONS: Our data clearly showed the differential expression of 11 miRNAs in cis-platin resistant cells, which could potentially target many important pathways including MAPK, TGF-? signaling, actin cytoskeleton, ubiquitin mediated proteasomal pathway, Wnt signaling, mTOR signaling, Notch signaling, apoptosis, and many other signaling pathways. Manipulation of one or more of these miRNAs could be an important approach for ovarian cancer chemotherapy.
Project description:Dumbbell-like Au-Fe(3)O(4) nanoparticles (NPs) were made and coupled with Herceptin and a platin complex. The platin-Au-Fe(3)O(4)-Herceptin NPs act as a target-specific nanocarriers for delivery of platin into Her2-positive breast cancer cells (Sk-Br3) with strong therapeutic effects. The conjugate has a half-maximal inhibitory concentration (IC(50)) toward Sk-Br3 cells of 1.76 microg of Pt/mL, which is lower than that needed for cisplatin (3.5 microg/mL). The work demonstrates that the dumbbell-like Au-Fe(3)O(4) NPs are promising nanocarriers for highly sensitive diagnostic and therapeutic applications.
Project description:Lysine histone demethylases (KDMs) are considered potential therapeutic targets in several tumors, including glioblastoma (GB). In particular, KDM5A is involved in the acquisition of temozolomide (TMZ) resistance in adult GB cells and UDX/KDM6B regulates H3K27 methylation, which is involved in the pediatric diffuse intrinsic pontine glioma (DIPG). Synthetic inhibitors of KDM5A (JIB 04 and CPI-455) efficiently block the proliferation of native and TMZ-resistant cells and the KDM6B inhibitor GSK J4 improves survival in a model of DIPG. The aim of our work was to determine if GSK J4 could be effective against GB cells that have acquired TMZ resistance and if it could synergize with TMZ or JIB 04 to increase the clinical utility of these molecules. Standard functional and pharmacological analytical procedures were utilized to determine the efficacy of the molecules under study when used alone or in combination against native GB cells and in a model of drug resistance. The results of this study indicated that although GSK J4 is active against native and TMZ-resistant cells, it does so at a lower efficacy than JIB 04. Drug combination studies revealed that GSK J4, differently from JIB 04, does not synergize with TMZ. Interestingly, GSK J4 and JIB 04 strongly synergize and are a potent combination against TMZ-resistant cells. Further studies in animal models will be necessary to determine if this combination of molecules might foster the development of novel therapeutic approaches for glioblastoma.