Project description:A metabolic hallmark of cancer identified by Warburg is the increased consumption of glucose and secretion of lactate, even in the presence of oxygen. Although many tumors exhibit increased glycolytic activity, most forms of cancer rely on mitochondrial respiration for tumor growth. We report here that Hürthle cell carcinoma of the thyroid (HTC) models harboring mitochondrial DNA-encoded defects in complex I of the mitochondrial electron transport chain exhibit impaired respiration and alterations in glucose metabolism. CRISPR-Cas9 pooled screening identified glycolytic enzymes as selectively essential in complex I-mutant HTC cells. We demonstrate in cultured cells and a PDX model that small molecule inhibitors of lactate dehydrogenase selectively induce an ATP crisis and cell death in HTC. This work demonstrates that complex I loss exposes fermentation as a therapeutic target in HTC and has implications for other tumors bearing mutations that irreversibly damage mitochondrial respiration.
Project description:A metabolic hallmark of cancer identified by Warburg is the increased consumption of glucose and secretion of lactate, even in the presence of oxygen. Although many tumors exhibit increased glycolytic activity, most forms of cancer rely on mitochondrial respiration for tumor growth. We report here that Hürthle cell carcinoma of the thyroid (HTC) models harboring mitochondrial DNA-encoded defects in complex I of the mitochondrial electron transport chain exhibit impaired respiration and alterations in glucose metabolism. CRISPR-Cas9 pooled screening identified glycolytic enzymes as selectively essential in complex I-mutant HTC cells. We demonstrate in cultured cells and a PDX model that small molecule inhibitors of lactate dehydrogenase selectively induce an ATP crisis and cell death in HTC. This work demonstrates that complex I loss exposes fermentation as a therapeutic target in HTC and has implications for other tumors bearing mutations that irreversibly damage mitochondrial respiration.
Project description:In this study we used unbiased approach in the lung cancer and colon cell lines (A549 and HTC 116 respectively) to identify universal early transcriptomic signatures of C-1305 cytotoxicity, and to highlight novel pathways responsible for its biological activity. The data obtained with real time analysis was used to select appropriate doses for subsequent RNAseq and biochemical analysis. Furthermore, the RNA samples prior RNA-seq analysis were pre-verified for transcriptomic activation of apoptosis related pathways via qPCR . Since our real time analysis of cell growth have shown that 24 h exposure to C-1305 (at IC50 concentrations) is sufficient to significantly alter A549 and HTC 116 cells growth and to activate apoptosis-related transcriptional signals, we determined genome wide transcriptomic alterations in A549 and HTC 116 upon C-1305 treatment. In brief, A549 and HTC 116 cells were exposed to 3 µM or 10 µM of C-1305, respectively for 24 h day after plating. Furthermore, since our data have shown that 24 h exposure of noncancer cells epithelial lung cells 16HBE14o- to 3 µM C-1305 resulted in minimal cellular damage and loss of viability, we have included this treatment of 16HBE14o- as a negative control in our analysis. Finally, since genomic instabilities were reported for A549 cells to avoid related artifacts, DMSO vehicle treated cells (controls) were obtained after 24 h and after 8 h of cell culture. Total RNA was extracted from the cells after 24 h exposure to C-1305 and 24 h and/or 8 h exposure to vehicle and subjected to RNA sequencing.
Project description:Wor1 is a conserved fungal regulatory protein that control the phenotypic switching and pathogenicity of Candida albicans.We simulated three models of Beauveria bassiana morphological transitions, including CTH (conidia to hyphae), HTC (hyphae to conidia) and BTB (blastospore to blastospore).We find that disruption of BbWor1 (an ortholog of Wor1) resulted in a distinct reduction in the time required for germination (CTH), a significant increase in hyphal growth and a decrease in the yield of conidia (HTC), indicating that BbWor1 positively controls conidia production and negatively regulates hyphal growth in conidia-hyphae switching. Meanwhile, the ΔBbWor1 prominently decreased blastospore yield, as well as shortening G0/G1 phase and prolonging G2/M phase, under the BTB model. Importantly, BbWor1 contributed to conidia-hyphae switching and blastospore propagation via different genetic pathways, and yeast one-hybrid testing demonstrated the necessity of BbWor1 to control the transcription of an allergen-like protein gene (BBA_02580), as well as a conidial wall protein gene (BBA_09998). Moreover, the dramatically weakened virulence of ΔBbWor1 was examined by immersion and injection methods.