Search

We describe a protocol for the discovery of synergistic drug combinations for the treatment of disease. Synergistic drug combinations lead to the use of drugs at lower doses, which reduces side effects and can potentially lead to reduced drug resistance, while being clinically more effective than the individual drugs. To cope with the extremely large search space for these combinations, we developed an efficient combinatorial drug screening method called the Feedback System Control (FSC) technique. Starting with a broad selection of drugs, the method follows an iterative approach of experimental testing in a relevant bioassay and analysis of the results by FSC. First, the protocol uses a cell viability assay to generate broad dose-response curves to assess the efficacy of individual compounds. These curves are then used to guide the dosage input of each drug to be tested in combination. Data from applied drug combinations are input into the differential evolution (DE) algorithm, which predicts new combinations to be tested in vitro. This process identifies optimal drug-dose combinations, while saving orders of magnitude in experimental effort. The complete optimization process is estimated to take ∼4 weeks. FSC does not require insight into the disease mechanism, and it has therefore been applied to find combination therapies for many different pathologies, including cancer and infectious diseases, and it has also been used in organ transplantation.

A major key to improvement of cancer therapy is the combination of drugs. Mixing drugs that already exist on the market may offer an attractive alternative. Here we report on a new model-based streamlined feedback system control (s-FSC) method, based on a design of experiment approach, for rapidly finding optimal drug mixtures with minimal experimental effort. We tested combinations in an in vitro assay for the viability of a renal cell adenocarcinoma (RCC) cell line, 786-O. An iterative cycle of in vitro testing and s-FSC analysis was repeated a few times until an optimal low dose combination was reached. Starting with ten drugs that target parallel pathways known to play a role in the development and progression of RCC, we identified the best overall drug combination, being a mixture of four drugs (axitinib, erlotinib, dasatinib and AZD4547) at low doses, inhibiting 90% of cell viability. The removal of AZD4547 from the optimized drug combination resulted in 80% of cell viability inhibition, while still maintaining the synergistic interaction. These optimized drug combinations were significantly more potent than monotherapies of all individual drugs (p

Drug combinations can improve angiostatic cancer treatment efficacy and enable the reduction of side effects and drug resistance. Combining drugs is non-trivial due to the high number of possibilities. We applied a feedback system control (FSC) technique with a population-based stochastic search algorithm to navigate through the large parametric space of nine angiostatic drugs at four concentrations to identify optimal low-dose drug combinations. This implied an iterative approach of in vitro testing of endothelial cell viability and algorithm-based analysis. The optimal synergistic drug combination, containing erlotinib, BEZ-235 and RAPTA-C, was reached in a small number of iterations. Final drug combinations showed enhanced endothelial cell specificity and synergistically inhibited proliferation (p < 0.001), but not migration of endothelial cells, and forced enhanced numbers of endothelial cells to undergo apoptosis (p < 0.01). Successful translation of this drug combination was achieved in two preclinical in vivo tumor models. Tumor growth was inhibited synergistically and significantly (p < 0.05 and p < 0.01, respectively) using reduced drug doses as compared to optimal single-drug concentrations. At the applied conditions, single-drug monotherapies had no or negligible activity in these models. We suggest that FSC can be used for rapid identification of effective, reduced dose, multi-drug combinations for the treatment of cancer and other diseases.

The cell is a complex system involving numerous components, which may often interact in a non-linear dynamic manner. Diseases at the cellular level are thus likely to involve multiple cellular constituents and pathways. As some drugs, or drug combinations, may act synergistically on these multiple pathways, they might be more effective than the respective single target agents. Optimizing a drug mixture for a given disease in a particular patient is particularly challenging due to both the difficulty in the selection of the drug mixture components to start out with, and the all-important doses of these drugs to be applied. For n concentrations of m drugs, in principle, n(m) combinations will have to be tested. As this may lead to a costly and time-consuming investigation for each individual patient, we have developed a Feedback System Control (FSC) technique which can rapidly select the optimal drug-dose combination from the often millions of possible combinations. By testing this FSC technique in a number of experimental systems representing different disease states, we found that the response of cells to multiple drugs is well described by a low order, rather smooth, drug-mixture-input/drug-effect-output multidimensional surface. The main consequences of this are that optimal drug combinations can be found in a surprisingly small number of tests, and that translation from in vitro to in vivo is simplified. This points to the possibility of personalized optimal drug mixtures in the near future. This unexpectedly simple input-output relationship may also lead to a simple solution for handling the issue of human diversity in cancer therapeutics.

Background and objectivesHelicobacter pylori (HP) is an endemic pathogenic bacterium causing gastritis and gastroduodenal ulceration in humans and is linked to the development of gastric malignancies. These first human in vivo studies investigated the photoeradication of HP using laser and white light.Study design/materials and methodsIn 13 HP-positive volunteers, a zone of gastric antrum was irradiated with laser (410 nm, 50 J/cm(2)) or endoscopic white light (10 J/cm(2)) 45 minutes after oral 5-aminolevulinic acid (5-ALA) 20 mg/kg. HP-eradication was assessed by biopsy urease test and HP-culture from irradiated and control zones 5 minutes, 4 and 48 hours post-irradiation.ResultsA maximum eradication effect was achieved at 4 hours post-irradiation when 85% of biopsies in the monochromatic and 66% in the white light exposed zones, and 58 and 33% in the respective control zones were HP-negative.ConclusionsHP numbers were greatly reduced following exposure to 5-ALA and either laser or white light in vivo. Photoeradication appears feasible, but further light dosimetry and the development of convenient application methods is required.

The Jungfraujoch Research Station (46.55[degrees]N, 7.98[degrees]E, 3580 m above sea level) for decades has contributed in a significant manner to the systematic observation of the Earth's atmosphere both with in situ measurements and with trace gas column detection. We report on the development of a lidar system that improves the measurement potential of highly resolved atmospheric parameters in both time and space, with the goal of achieving long-term monitoring of atmospheric aerosol optical properties and water-vapor content. From the simultaneously detected elastic-backscatter signals at 355, 532, and 1064 nm, Raman signals from nitrogen at 387 and 607 nm, and water vapor at 408 nm, the aerosol extinction and backscatter coefficients at three wavelengths and a water-vapor mixing ratio are derived. Additional information about particle shape is obtained by depolarization measurements at 532 nm. Water-vapor measurements by use of both nitrogen and water-vapor Raman returns from the 355-nm laser beam are demonstrated with a vertical range resolution of 75 m and an integration time of 2 h. The comparison to the water-vapor profile derived from balloon measurements (Snow White technique) showed excellent agreement. The system design and the results obtained by its operation are reported. OCIS codes: 010.3640, 010.1100, 010.7030.

Numerical simulations compared with field measurements are used to explain the effect of sea breezes on photochemical smog episodes in Athens during the Mediterranean Campaign of Photochemical Tracers on 1214 September 1994. The numerical simulations, performed using a nonhydrostatic vorticity mesoscale model coupled to the Lurmann-Carter-Coyner photochemical module, are compared with ground-based lidar and aircraft measurements. The current analysis shows that the three selected days include the two main summertime flow patterns characteristic of the Athens peninsula, each of which lead to significantly different pollution amounts. On 12 and 13 September, a strong, northerly synoptic wind reduces the inland penetration of the sea breeze so that ozone concentrations within the greater Athens area remained low. In contrast, the weaker synoptic forcing on 14 September allowed the development of sea breezes over the whole peninsula and high ozone concentrations were found north and east of the city. An analysis based on pollution amounts and wind patterns is carried out to divide the peninsula into regions, each of which corresponds to a specific pollutant behavior.

From lidar signals detected with a shot per shot differential absorption lidar instrument tuned for tropospheric ozone measurements and recording each individual return, we reconstruct histograms of their sampled values for each channel of our digitizers. The analysis of their shape permits the correction of our measurements for experimental biases. In particular, a negative correlation is found between the skew of the histograms and the intensity of the backscattered light. The skew comes from a tail at high sampled values, interpreted as due to a raise of the relative contribution to the signal of a signal-induced noise when this intensity diminishes. By fitting a Gaussian function to the histograms without considering their tails, we calculate average signals unbiased by the corresponding noise. This approach allows us to increase the range of our ozone profiles, up to as much as double it in some cases. Key words: Lidar, ozone, troposphere, atmospheric fluctuations, signal-induced bias.

A fluorescence imaging device applied to the detection of early cancer is described. The apparatus is based on the imaging of laser-induced fluorescence of a dye that localizes in a tumor with a higher concentration than in the surrounding normal tissue after iv injection. Tests carried out in the upper aerodigestive tract, the tracheobronchial tree, and the esophagus with Photofrin II (1 mg/kg of body weight) as the fluorescent agent are reported as examples. The fluorescence is induced by violet (410-nm) light from a continuous-wave (cw) krypton-ion laser. The fluorescence contrast between tumor and surrounding tissue is enhanced by real-time image processing. This is done by the simultaneous recording of the fluorescence image in two spectral domains (470-600 and 600-720 nm), after which these two images are digitized and manipulated with a mathematical operator (look-up table) at video frequency. Among the 7 photodetections performed in the tracheobronchial tree, 6 were successful, whereas it was the case for only 5 of the 15 lesions investigated in squamous mucosa (upper aerodigestive tract and esophagus). The sources of false positives and false negatives are evaluated in terms of the fluorescent dye, tissue optical properties, and illumination optics. Key words: Laser-induced fluorescence, Photofrin II, fluorescence, imaging, cancer, photodetection, hematoporphyrin derivative, tissue diagnosis.

Photochemical incineration of anthracene was conducted to investigate proper disposal techniques for toxic and municipal waste. A flowing gas experiment was undertaken, where anthracene vapor was diluted in 1 atm of filtered compressed air and flowed through the all-quartz cell of 50 cm. length and 38 mm. diameter. Results showed the thermal and the photon-assisted thermal destruction of anthracene.