Project description:Mithramycin A in known to bind DNA but its exact cellular mechanism of action is still unclear. We used Affymatrix GenFlex_Tag_16K_V2 microarrays to profile sensitivity of genetically barcoded S. cerevisiae gene deletion strains to mithramycin A
Project description:We used microarrays to detail the global gene expression of mithramycin treated cells, xenografts and sp1 depleted cells with p53 overexpression. MPM cells and xenografts were treated with mithramycin or SP1 depleted/p53 overexpressed MPM cells were selected for RNA extraction and hybridization on Affymetrix microarrays. We sought to see the invitro and invivo effect of mithramycin (pharmacological model) and depletion of SP1 and overexpression of p53 (genetic model).
Project description:The study aims to define gene expression changes associated with mithramycin treatment of Ewing Sarcoma cell lines. The data consist of 12 arrays. Two cell lines, TC71 and TC32, were treated with solvent control or with mithramycin, and RNA was extracted at 6 hours. Three biological replicates per cell line/treatment.
Project description:Yeast homozygous and essential heterozygous deletion mutants were screened against 3,250 diverse growth inhibitory compounds, resulting in a chemogenomic profile for each compound. A profile permits the identification of putative protein targets via drug-induced haploinsufficiency, and provides a genome-wide description of the cellular response to the profiled compound. Therefore, our aims were to identify putative protein inhibitors and in general, to improve understanding of the cellular response to small molecules.
Project description:Breast cancer is a heterogeneous collection of disease arising from the breast with distinct molecular and phenotypic features. Certain subsets of patients have tumors that are particularly difficult-to-treat, which include triple-negative breast cancer (TNBC), metastatic/recurrent disease and rare histological variants. To delineate the underlying biology and identify therapeutic candidates for these patients, a series of 37 breast cancer patient-derived xenografts (PDX) from both chemo-naïve and pre-treated specimens was generated from 81 transplant attempts. Whole-genome and transcriptome sequencing revealed marked fidelity of the molecular landscape for the majority of PDXs in comparison to parental tumors. Reverse-phase protein array analysis of PDXs further identified potential therapeutic targets. Metastatic potential varied between PDXs, where low-penetrance lung micrometastases was the most common pattern of dissemination and observed in 34.5% of models. Three PDXs recapitulated the metastatic localization seen in the corresponding patients, including two lines with high-frequency metastases to multiple vital organ systems, while another PDX displayed tropism to the skull-base. Chemosensitivity profiling was performed in vivo with standard-of-care agents (doxorubicin, cisplatin, gemcitabine or paclitaxel), where multi-drug chemoresistance was found in 60.0% of PDXs and 64.7% of responses were concordant with pre-engraftment responses observed in the patient. Consolidating chemogenomic data identified potentially actionable features in 97.2% of PDXs, and marked regressions were seen in vivo when a subset of these underwent proof-of-concept functional studies. This included FGFR inhibitor sensitivity in a FGFR1-amplified lobular carcinoma, mTOR inhibitor sensitivity in a recurrent neuroendocrine breast cancer with proteomic evidence of mTOR/PI3K activation, and platinum sensitivity in a TNBC with BRCA1 germline mutation predicted as benign. Together, this clinically-annotated PDX library with comprehensive molecular and phenotypic profiling serves as a resource for both discovery and validation preclinical studies on difficult-to-treat breast tumors.
Project description:Transcriptional profiling of primary human malignant B cells from patients with chronic lymphocytic leukaemia (CLL), comparing untreated CLL cells with CLL cells that have been treated 24h with 200nM mithramycin. Mithramycin intercalates into GC-rich regions of DNA to inhibit binding of the transcription factor SP1. The goal of this experiment is to gain understanding of the genes regulated by SP1 in primary CLL cells.
