ABSTRACT
γ-Secretase mediates amyloid production in Alzheimer's disease (AD) and oncogenic activity of Notch. γ-Secretase inhibitors (GSIs) are thus of interest for AD and oncology. A peripheral biomarker of Notch activity would aid determination of the therapeutic window and dosing regimen for GSIs, given toxicities associated with chronic Notch inhibition. This study examined the effects of GSI MK-0752 on blood and hair follicle transcriptomes in healthy volunteers. The effects of a structurally diverse GSI on rhesus blood and hair follicles were also compared. Significant dose-related effects of MK-0752 on transcription were observed in hair follicles, but not blood. The GSI biomarker identified in follicles exhibited 100% accuracy in a clinical test cohort, and was regulated in rhesus by a structurally diverse GSI. This study identified a translatable, accessible pharmacodynamic biomarker of GSI target engagement and provides proof of concept of hair follicle RNA as a translatable biomarker source.
Subject(s)
Amyloid Precursor Protein Secretases/antagonists & inhibitors , Benzene Derivatives/pharmacology , Drug Monitoring , Hair Follicle/drug effects , Propionates/pharmacology , Protease Inhibitors/pharmacology , Receptors, Notch/antagonists & inhibitors , Sulfones/pharmacology , Transcription, Genetic/drug effects , Adolescent , Adult , Amyloid Precursor Protein Secretases/metabolism , Animals , Baltimore , Benzene Derivatives/administration & dosage , Benzene Derivatives/blood , Benzene Derivatives/pharmacokinetics , Biomarkers, Pharmacological/blood , Cross-Over Studies , Dose-Response Relationship, Drug , Double-Blind Method , Drug Monitoring/methods , Gene Expression Profiling/methods , Hair Follicle/metabolism , Healthy Volunteers , Humans , Macaca mulatta , Male , Models, Animal , Molecular Targeted Therapy , Oligonucleotide Array Sequence Analysis , Propionates/administration & dosage , Propionates/blood , Propionates/pharmacokinetics , Protease Inhibitors/administration & dosage , Protease Inhibitors/blood , Protease Inhibitors/pharmacokinetics , RNA, Messenger/biosynthesis , RNA, Messenger/blood , Receptors, Notch/metabolism , Sulfones/administration & dosage , Sulfones/blood , Sulfones/pharmacokinetics , Young AdultABSTRACT
Glioblastoma multiforme is the most common and lethal primary malignant brain tumor. Although considerable progress has been made in technical proficiencies of surgical and radiation treatment for brain tumor patients, the impact of these advances on clinical outcome has been disappointing, with median survival time not exceeding 15 months. Over the last 30 years, no significant increase in survival of patients suffering from this disease has been achieved. A fundamental source of the management challenge presented in glioma patients is the insidious propensity of tumor invasion into distant brain tissue. Invasive tumor cells escape surgical removal and geographically dodge lethal radiation exposure and chemotherapy. Recent improved understanding of biochemical and molecular determinants of glioma cell invasion provide valuable insight into the underlying biological features of the disease, as well as illuminating possible new therapeutic targets. These findings are moving forward to translational research and clinical trials as novel antiglioma therapies.
Subject(s)
Glioma/metabolism , Glioma/pathology , Neoplasm Invasiveness/pathology , Clinical Trials as Topic , Extracellular Matrix/metabolism , Glioma/therapy , Humans , Neoplasm Invasiveness/prevention & control , Neoplasm Proteins/metabolism , Neoplastic Cells, Circulating/pathologyABSTRACT
Microarray analysis of complementary DNA (cDNA) allows large-scale, comparative, gene expression profiling of two different cell populations. This approach has the potential for elucidating the primary transcription events and genetic cascades responsible for increased glioma cell motility in vitro and invasion in vivo. These genetic determinants could become therapeutic targets. We compared cDNA populations of a glioma cell line (G112) exposed or not to a motility-inducing substrate of cell-derived extracellular matrix (ECM) proteins using two sets of cDNA microarrays of 5,700 and 7,000 gene sequences. The data were analyzed considering the level and consistency of differential expression (outliers) and whether genes involved in pathways of motility, apoptosis, and proliferation were differentially expressed when the motility behavior was engaged. Validation of differential expression of selected genes was performed on additional cell lines and human glioblastoma tissue using quantitative RT-PCR. Some genes involved in cell motility, like tenascin C, neuropilin 2, GAP43, PARG1 (an inhibitor of Rho), PLCy, and CD44, were over expressed; other genes, like adducin 3y and integrins, were down regulated in migrating cells. Many key cell cycle components, like cyclin A and B, and proliferation markers, like PCNA, were strongly down regulated on ECM. Interestingly, genes involved in apoptotic cascades, like Bcl-2 and effector caspases, were differentially expressed, suggesting the global down regulation of proapoptotic components in cells exposed to cell-derived ECM. Overall, our findings indicate a reduced proliferative and apoptotic activity of migrating cells. cDNA microarray analysis has the potential for uncovering genes linking the phenotypic aspects of motility, proliferation, and apoptosis.
Subject(s)
Brain Neoplasms/pathology , Gene Expression Regulation, Neoplastic , Glioma/pathology , Neoplasm Invasiveness/genetics , Neoplasm Proteins/biosynthesis , Transcription, Genetic , Apoptosis/genetics , Brain Neoplasms/chemistry , Cell Cycle Proteins/biosynthesis , Cell Cycle Proteins/genetics , Cell Movement/drug effects , Cell Movement/genetics , Computer Systems , Culture Media/pharmacology , DNA, Complementary/genetics , Expressed Sequence Tags , Extracellular Matrix Proteins/pharmacology , Gene Expression Profiling , Gene Expression Regulation, Neoplastic/drug effects , Glioblastoma/chemistry , Glioblastoma/pathology , Growth Substances/biosynthesis , Growth Substances/genetics , Humans , Lasers , Neoplasm Proteins/genetics , Oligonucleotide Array Sequence Analysis , Phenotype , Polymerase Chain Reaction , Tenascin/biosynthesis , Tenascin/genetics , Transcription, Genetic/drug effects , Tumor Cells, Cultured/drug effects , Tumor Cells, Cultured/metabolism , Tumor Cells, Cultured/pathologyABSTRACT
Two well-defined oxidative chlorination-cyclization processes have been developed for the stereoselective synthesis of a variety of 4-amido-isothiazolidinone oxide derivatives. The stereochemistry of the cyclization products was confirmed by X-ray crystallography. These new compounds were designed as bacterial serine protease inhibitors. In tests, some of them showed weak antibacterial activity.
Subject(s)
Amides/chemical synthesis , Anti-Bacterial Agents/chemical synthesis , Thiazoles/chemical synthesis , Amides/chemistry , Amides/pharmacology , Anti-Bacterial Agents/chemistry , Anti-Bacterial Agents/pharmacology , Chlorine/chemistry , Cyclization , Enterobacter cloacae/drug effects , Klebsiella pneumoniae/drug effects , Microbial Sensitivity Tests , Molecular Conformation , Moraxella catarrhalis/drug effects , Thiazoles/chemistry , Thiazoles/pharmacologyABSTRACT
A new migration assay, the time-lapse individual cell migration assay (TIM-assay), was developed, which allows the observation of cells over 24 h under controlled conditions. Using this technique, the migratory behavior of 8 human glioblastoma cell lines in vitro was studied. Special features are simultaneous documentation of migratory parameters of individual cells, i.e., migration velocities and migration paths of individual cells. Migration velocity for cell populations of the same cell line ranged from 0 to 24 microm/h. The migration paths were examined for being directional. Two thirds of all cells showed directional migration. Migration paths were further classified according to visual judgements for being linear, oscillating or mixed. The migration index had a mean of 91%. The presented TIM-assay allows the assessment of several new parameters. that may be useful to identify subgroups of gliomas with different biological characteristics.
Subject(s)
Brain Neoplasms/pathology , Cell Movement , Glioma/pathology , Microscopy, Video/methods , Adult , Aged , Female , Homeostasis , Humans , Hydrogen-Ion Concentration , Kinetics , Male , Neoplasm Invasiveness , Osmolar Concentration , Temperature , Tumor Cells, CulturedABSTRACT
This reports the synthesis and in vitro antimicrobial properties of a series of 2-thioether-linked quinolonyl-carbapenems. Although the title compounds exhibited broad spectrum activity, the MICs were generally higher than those observed for selected benchmark carbapenems, quinolonyl-penems, and quinolones. Enzyme assays suggested that the title compounds are potent inhibitors of penicillin binding proteins and inefficient inhibitors of bacterial DNA-gyrase. Uptake studies indicated that the new compounds are not substrates for the norA encoded quinolone efflux pump.
Subject(s)
Carbapenems/chemistry , Carbapenems/pharmacology , Gram-Negative Bacteria/drug effects , Gram-Positive Bacteria/drug effects , Quinolones/chemistry , Bacterial Proteins/drug effects , Carbapenems/chemical synthesis , Carrier Proteins/drug effects , Cell Division , Gram-Negative Bacteria/enzymology , Gram-Positive Bacteria/enzymology , Hexosyltransferases/drug effects , Microbial Sensitivity Tests , Multidrug Resistance-Associated Proteins , Multienzyme Complexes/drug effects , Muramoylpentapeptide Carboxypeptidase/drug effects , Penicillin-Binding Proteins , Peptidyl Transferases/drug effects , Structure-Activity Relationship , Topoisomerase II InhibitorsSubject(s)
Anti-Bacterial Agents/chemistry , Gram-Negative Bacteria/drug effects , Gram-Positive Bacteria/drug effects , Quinolones/chemistry , beta-Lactam Resistance , Animals , Anti-Bacterial Agents/pharmacology , Anti-Bacterial Agents/therapeutic use , Bacteria/drug effects , Drug Design , Humans , Lactams , Microbial Sensitivity Tests , Molecular Structure , Quinolones/pharmacology , Quinolones/therapeutic useABSTRACT
A series of cephalosporins derived from cephalothin containing an ester-linked quinolonyl substituent at the C-10 position (C-10 quinolonyl-cephem esters) has been prepared and evaluated for in vitro antibacterial activity. The C-10 quinolonyl-cephem esters exhibited a broadened spectrum of activity when compared with cephalothin and the corresponding quinolones, including activity against beta-lactamase-producing bacteria.
