Addressing the safety aspects of drugs and environmental chemicals has historically been undertaken through animal testing. However, the quantity of chemicals in need of assessment and the challenges of species extrapolation require the development of alternative approaches. Our approach, the US Environmental Protection Agency's ToxCast program, utilizes a large suite of in vitro and model organism assays to interrogate important chemical libraries and computationally analyze bioactivity profiles. Here we evaluated one component of the ToxCast program, the use of primary human cell systems, by screening for chemicals that disrupt physiologically important pathways. Chemical-response signatures for 87 endpoints covering molecular functions relevant to toxic and therapeutic pathways were generated in eight cell systems for 641 environmental chemicals and 135 reference pharmaceuticals and failed drugs. Computational clustering of the profiling data provided insights into the polypharmacology and potential off-target effects for many chemicals that have limited or no toxicity information. The endpoints measured can be closely linked to in vivo outcomes, such as the upregulation of tissue factor in endothelial cell systems by compounds linked to the risk of thrombosis in vivo. Our results demonstrate that assaying complex biological pathways in primary human cells can identify potential chemical targets, toxicological liabilities and mechanisms useful for elucidating adverse outcome pathways.

The complexity of human biology has made prediction of health effects as a consequence of exposure to environmental chemicals especially challenging. Complex cell systems, such as the Biologically Multiplexed Activity Profiling (BioMAP) primary, human, cell-based disease models, leverage cellular regulatory networks to detect and distinguish chemicals with a broad range of target mechanisms and biological processes relevant to human toxicity. Here the authors use the BioMAP human cell systems to characterize effects relevant to human tissue and inflammatory disease biology following exposure to the 320 environmental chemicals in the Environmental Protection Agency's (EPA's) ToxCast phase I library. The ToxCast chemicals were assayed at 4 concentrations in 8 BioMAP cell systems, with a total of 87 assay endpoints resulting in more than 100,000 data points. Within the context of the BioMAP database, ToxCast compounds could be classified based on their ability to cause overt cytotoxicity in primary human cell types or according to toxicity mechanism class derived from comparisons to activity profiles of BioMAP reference compounds. ToxCast chemicals with similarity to inducers of mitochondrial dysfunction, cAMP elevators, inhibitors of tubulin function, inducers of endoplasmic reticulum stress, or NFkappaB pathway inhibitors were identified based on this BioMAP analysis. This data set is being combined with additional ToxCast data sets for development of predictive toxicity models at the EPA.

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Microsites related to microenvironmental conditions, including microclimate, seem to be a key factor for the restoration of forests in the subalpine area. Tree growth was studied in Picea abies (L.) Karst. (Norway spruce) and Larix decidua Mill. (European larch) on 30 plots located at different microsites (i.e., different elevations and micro top o graphies combined) within the subalpine zone (1680–1940 m) of the Schmirn Valley (Tyrol, Austria). The age of the trees studied was 27 years for larch and 28 years for spruce. The mean height and biomass growth decreased significantly with increasing elevation. The effect of elevation and microtopography on growth varied with tree size (age): (1) elevation had little effect on growth of trees less than 0.5 m in height; (2) both elevation and microtopography affected tree growth significantly when the tree height was between 0.5 and 3 m; (3) as trees exceed 3 m in height, tree canopies can fully cover the ground surface and create a forest microclimate causing growth to decline with increasing elevation, irrespective of microtopography. We conclude that the microclimate, associated with microsite, controls growth during the early stages of tree development, but following canopy closure, the local climate (mesoclimate) associated with topography begins to determine tree growth.

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Compound mechanism-of-action information can be critical for drug development decisions but is often challenging for phenotypic drug discovery programs. One concern is that compounds selected by phenotypic screening will have a previously known but undesirable target mechanism. Here we describe a useful method for assigning mechanism class to compounds and bioactive agents using an 84-feature signature from a panel of primary human cell systems (BioMAP systems). For this approach, a reference data set of well-characterized compounds was used to develop predictive models for 28 mechanism classes using support vector machines. These mechanism classes encompass safety and efficacy-related mechanisms, include both target-specific and pathway-based classes, and cover the most common mechanisms identified in phenotypic screens, such as inhibitors of mitochondrial and microtubule function, histone deacetylase, and cAMP elevators. Here we describe the performance and the application of these predictive models in a decision scheme for triaging phenotypic screening hits using a previously published data set of 309 environmental chemicals tested as part of the Environmental Protection Agency's ToxCast program. By providing quantified membership in specific mechanism classes, this approach is suitable for identification of off-target toxicity mechanisms as well as enabling target deconvolution of phenotypic drug discovery hits.

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-We examined growth in Pinus cembra L. (28 years old) across the treeline ecotone from 1900 to 2100 m elevation in the Alps. Eighteen plots were chosen at different microsites which are defined as a combination of elevation and steepness (gentle vs. steep slope) on a south-facing slope in the Schmirn Valley, Tyrol/Austria. Over the range of altitudes studied, elevation and steepness had influences on growth depending on tree size: (1) Elevation and steepness had little effect on growth as long as trees were very small (< 0.5 m in height); (2) Both elevation and steepness affected tree growth significantly when the tree height was between 0.5 and 3 m; (3) As trees exceeded 3 m in height, tree canopies fully covered the ground surface and created a forest microclimate causing growth to decline with increasing elevation, irrespective of steepness. We conclude that the microsite related to microclimate, controls growth during the early life stages of trees, but following canopy closure the local climate (mesoclimate) associated with topography begins to determine tree growth. growth responses / high altitude / micro-environmental conditions / tree ecology / treeline ecotone Résumé -Effets de la microstation sur la croissance de Pinus cembra dans la zone subalpine des Alpes autrichiennes. Nous avons examiné la croissance de Pinus cembra L. (28 ans) dans l'écotone de la limite forestière entre 1900 et 2100 m d'altitude dans les Alpes. Dixhuit placettes ont été choisies dans différentes microstations définies selon l'altitude et la déclivité du terrain sur une pente exposée au sud dans la vallée de Schmirn, dans le Tyrol autrichien. Dans toute la zone étudiée, l'altitude et la déclivité ont exercé une influence qui dépendait de la taille de l'arbre: (1) elles avaient peu d'effet sur la croissance des arbres de très petite taille (< 0.5 m de haut); (2) elles avaient un effet significatif sur les arbres d'une hauteur entre 0,5 et 3 m; (3) à partir de 3 m de haut, la canopée couvrait complètement la surface du sol et créait ainsi un microclimat forestier qui entraîne un ralentissement de la croissance avec l'augmentation de l'altitude, indépendamment de la déclivité. Nous en concluons que la microstation liée au microclimat détermine la croissance des arbres durant leur jeune âge, mais après la fermeture de la canopée, le climat local (mésoclimat) associé à la topographie commence à influencer la croissance des arbres. réactions à la croissance / altitude / conditions microenvironnementales / écologie des arbres / écotone de la limite forestière