Project description:Cellular transitions hold great promise in translational medicine research. However, therapeutic applications are limited by the low efficiency and safety concerns of using transcription factors. Small molecules provide a temporal and highly tuneable approach to overcome these issues. Here, we present PC3T, a computational framework to identify enrich molecules that induce desired cellular transitions, and PC3T was able to consistently identify enrich small molecules that had been experimentally validated in both bulk and single cell datasets. We then predicted and experimentally validated small molecules reprogramming of fibroblasts into hepatocyte-like cells and found that the fibroblasts exhibited epithelial cell morphology and hepatocyte-like specific gene expression pattern after treatment.
Project description:The declining health of coral reefs worldwide is likely to intensify in response to continued anthropogenic disturbance from coastal development, pollution, and climate change. In response to these stresses, reef-building corals may exhibit bleaching, which marks the breakdown in symbiosis between coral and zooxanthellae. Mass coral bleaching due to elevated water temperature can devastate coral reefs on a large geographic scale. In order to understand the molecular and cellular basis of bleaching in corals, we have measured gene expression changes associated with thermal stress and bleaching using a cDNA microarray containing 1,310 genes of the Caribbean coral Montastraea faveolata. In a first experiment, we identified differentially expressed genes by comparing experimentally bleached M. faveolata fragments to control non-heat-stressed fragments. We also identified differentially expressed genes during a time course experiment with four time points across nine days. Results suggest that thermal stress and bleaching in M. faveolata affect the following processes: oxidative stress, Ca2+ homeostasis, cytoskeletal organization, cell death, calcification, metabolism, protein synthesis, heat shock protein activity, and transposon activity. These results represent the first large-scale transcriptomic study focused on revealing the cellular foundation of thermal stress-induced coral bleaching. We postulate that oxidative stress in thermal-stressed corals causes a disruption of Ca2+ homeostasis, which in turn leads to cytoskeletal and cell adhesion changes, decreased calcification, and the initiation of cell death via apoptosis and necrosis. Keywords: thermal stress response, time course, coral bleaching
Project description:The declining health of coral reefs worldwide is likely to intensify in response to continued anthropogenic disturbance from coastal development, pollution, and climate change. In response to these stresses, reef-building corals may exhibit bleaching, which marks the breakdown in symbiosis between coral and zooxanthellae. Mass coral bleaching due to elevated water temperature can devastate coral reefs on a large geographic scale. In order to understand the molecular and cellular basis of bleaching in corals, we have measured gene expression changes associated with thermal stress and bleaching using a cDNA microarray containing 1,310 genes of the Caribbean coral Montastraea faveolata. In a first experiment, we identified differentially expressed genes by comparing experimentally bleached M. faveolata fragments to control non-heat-stressed fragments. We also identified differentially expressed genes during a time course experiment with four time points across nine days. Results suggest that thermal stress and bleaching in M. faveolata affect the following processes: oxidative stress, Ca2+ homeostasis, cytoskeletal organization, cell death, calcification, metabolism, protein synthesis, heat shock protein activity, and transposon activity. These results represent the first large-scale transcriptomic study focused on revealing the cellular foundation of thermal stress-induced coral bleaching. We postulate that oxidative stress in thermal-stressed corals causes a disruption of Ca2+ homeostasis, which in turn leads to cytoskeletal and cell adhesion changes, decreased calcification, and the initiation of cell death via apoptosis and necrosis. Keywords: thermal stress response; coral bleaching
Project description:Toxoplasma gondii is a single-celled eukaryotic parasite that chronically infects a quarter of the global population. In recent years, phenotypic screens have identified compounds that block parasite replication. Unraveling the complexity of pathogenesis and molecular mechanisms and pathways perturbed by such compounds requires target deconvolution. Thermal proteome profiling (TPP), also known as the cellular thermal shift assay (CETSA), recently emerged as a method to identify small molecule–target interactions in living cells and cell extracts in a variety of organisms, including unicellular eukaryotic pathogens. Ligand binding induces a thermal stability shift—stabilizing or destabilizing proteins that change conformationally in response to the ligand—that can be measured by mass spectrometry (MS). Cells are incubated with different concentrations of ligand and heated, causing thermal denaturation of proteins. The soluble protein is extracted and quantified with multiplexed, quantitative MS, giving rise to thousands of thermal denaturation profiles. Proteins engaging the ligand can be identified by their concentration-dependent thermal shift. The protocol provided here can be used to identify compound-target interactions and assess the impact of environmental or genetic perturbation on the thermal stability of the proteom in T. gondii and other eukaryotic pathogens. Here, we generate a reference dataset of the thermal profiles of extracellular T. gondii tachyzoites.
Project description:Red fruits are valued for their vitamin C and polyphenol content, but traditional heat preservation methods used in juice and nectar production can significantly reduce these components. Therefore, alternative non-thermal methods are explored to inactivate foodborne pathogens like Escherichia coli while maintaining the nutritional value. However, knowledge about the effects of these technologies on bacterial cells is limited. This study analyzed differentially expressed genes of E. coli ATCC 8739 inoculated in strawberry nectar after exposure to three treatments with two sets of parameters each, namely thermal treatment, high-pressure processing (HPP), and moderate-intensity pulsed electric field (MIPEF). The highest inactivation efficiency was achieved with HPP at 400 MPa, 1 min, reducing microbial counts by 5.0±0.3 log cfu/mL, and thermal treatment at 60°C, 200 s, achieving a reduction of 4.4±0.2 log cfu/mL, while no inactivation was observed with MIPEF at 6 kV/cm. Transcriptomic analysis showed that thermal and HPP treatments caused similar molecular stress responses in E. coli. In both cases, the most overexpressed genes encoded outer membrane proteins, which may lead to the activation of the envelope stress response. Despite no microbial inactivation was revealed after MIPEF treatment, strong transcriptomic responses were observed, particularly in genes related to membrane integrity and metabolic activity. Numerous overexpressed genes associated with ABC transporters, outer membrane proteins, and lipoproteins were identified, which could increase the strain’s virulence. This study provides insights into the stress response mechanisms induced by conventional and novel treatments. Nevertheless, further research is needed to investigate the long-term effects on bacterial populations.
Project description:Here, we screened a library of non-characterized small molecules against CRC patient-derived tumor spheroid culture (TSC) cells to identify a compound (NCT02) with activity in a subset of TSCs. Transcriptomics as well as thermal proteome profiling and chemical proteomics identified degradation of cyclin K (CCNK) and CDK12 as central mechanism-of-action, associated with increase of DNA damage and induction of apoptosis.
Project description:Coral bleaching and coral reef degradation become severe as the surface seawater temperature rises. Much research to date has focused on the bacterial community composition properties within the coral holobiont, but less attention has been paid to the interactions of bacteria and corals under thermal stress. We investigated the changes of coral symbiotic bacteria and metabolites under thermal stress, and analyzed the internal relationship between bacteria and metabolites as well as their relationship with coral health. We found obvious signs of coral bleaching after heating treatment, and the interaction within symbiotic bacterial community became closer. The coral symbiotic bacterial community and metabolites changed significantly under thermal stress, and bacteria such as Flavobacterium, Shewanella and Psychrobacter increased significantly. Bacteria associated with stress tolerance, biofilm formation and mobile elements decreased, and bacterial DMSP metabolism increased slightly after heating treatment. Differential metabolites in corals after heating treatment were associated with cell cycle regulation and antioxidant. This study revealed the correlation between differential metabolites and bacterial community composition changes in corals under thermal stress, and providing valuable insight on metabolomics research of corals.
Project description:The declining health of coral reefs worldwide is likely to intensify in response to continued anthropogenic disturbance from coastal development, pollution, and climate change. In response to these stresses, reef-building corals may exhibit bleaching, which marks the breakdown in symbiosis between coral and zooxanthellae. Mass coral bleaching due to elevated water temperature can devastate coral reefs on a large geographic scale. In order to understand the molecular and cellular basis of bleaching in corals, we have measured gene expression changes associated with thermal stress and bleaching using a cDNA microarray containing 1,310 genes of the Caribbean coral Montastraea faveolata. In a first experiment, we identified differentially expressed genes by comparing experimentally bleached M. faveolata fragments to control non-heat-stressed fragments. We also identified differentially expressed genes during a time course experiment with four time points across nine days. Results suggest that thermal stress and bleaching in M. faveolata affect the following processes: oxidative stress, Ca2+ homeostasis, cytoskeletal organization, cell death, calcification, metabolism, protein synthesis, heat shock protein activity, and transposon activity. These results represent the first large-scale transcriptomic study focused on revealing the cellular foundation of thermal stress-induced coral bleaching. We postulate that oxidative stress in thermal-stressed corals causes a disruption of Ca2+ homeostasis, which in turn leads to cytoskeletal and cell adhesion changes, decreased calcification, and the initiation of cell death via apoptosis and necrosis. Keywords: thermal stress response, time course, coral bleaching Time course with 4 time points and 4 biological replicates per time point. Each biological replicate at each time point was hybridized to a pooled reference control sample containing RNA from all control non-heat-stressed coral fragments.
Project description:The declining health of coral reefs worldwide is likely to intensify in response to continued anthropogenic disturbance from coastal development, pollution, and climate change. In response to these stresses, reef-building corals may exhibit bleaching, which marks the breakdown in symbiosis between coral and zooxanthellae. Mass coral bleaching due to elevated water temperature can devastate coral reefs on a large geographic scale. In order to understand the molecular and cellular basis of bleaching in corals, we have measured gene expression changes associated with thermal stress and bleaching using a cDNA microarray containing 1,310 genes of the Caribbean coral Montastraea faveolata. In a first experiment, we identified differentially expressed genes by comparing experimentally bleached M. faveolata fragments to control non-heat-stressed fragments. We also identified differentially expressed genes during a time course experiment with four time points across nine days. Results suggest that thermal stress and bleaching in M. faveolata affect the following processes: oxidative stress, Ca2+ homeostasis, cytoskeletal organization, cell death, calcification, metabolism, protein synthesis, heat shock protein activity, and transposon activity. These results represent the first large-scale transcriptomic study focused on revealing the cellular foundation of thermal stress-induced coral bleaching. We postulate that oxidative stress in thermal-stressed corals causes a disruption of Ca2+ homeostasis, which in turn leads to cytoskeletal and cell adhesion changes, decreased calcification, and the initiation of cell death via apoptosis and necrosis. Keywords: thermal stress response; coral bleaching 5 control and 5 heat-stressed RNA samples were hybridized in a 5-replicate dye-swap design (10 total hyb's).
Project description:Cytochrome C from horse heart has became a standard molecule in terms of protein dynamics studies by following the exchange of amide protons at different positions in its seqeunce. Therefore both its structure and dynamics have been well characterised by different methods, like NMR or ETD-based MS. The present dataset presents a control classic HDX experiment which includes full deuteration control to be confronted with exchange rate mapping along the sequence obtained by other methods.