Quantitative detection of disseminated melanoma cells by trp-1 transcript analysis reveals stochastic distribution of pulmonary metastases

A better understanding of the process of melanoma metastasis is required to underpin the development of novel therapies that will improve patient outcomes. The use of appropriate animal models is indispensable for investigating the mechanisms of melanoma metastasis. However, reliable and practicable quantification of metastases in experimental mice remains a challenge, particularly if the metastatic burden is low. Here, we describe a qRT-PCR-based protocol that employs the melanocytic marker Trp-1 for the sensitive quantification of melanoma metastases in the murine lung. Using this protocol, we were able to detect the presence of as few as 100 disseminated melanoma cells in lung tissue. This allowed us to quantify metastatic burden in a spontaneous syngeneic B16-F10 metastasis model, even in the absence of visible metastases, as well as in the autochthonous Tg(Grm1)/Cyld$^{-/-}$ melanoma model. Importantly, we also observed an uneven distribution of disseminated melanoma cells amongst the five lobes of the murine lung, which varied considerably from animal to animal. Together, our findings demonstrate that the qRT-PCR-based detection of Trp-1 allows the quantification of low pulmonary metastatic burden in both transplantable and autochthonous murine melanoma models, and show that the analysis of lung metastasis in such models needs to take into account the stochastic distribution of metastatic lesions amongst the lung lobes

IER2-induced senescence drives melanoma invasion through osteopontin

Expression of the immediate-early response gene IER2 has been associated with the progression of several types of cancer, but its functional role is poorly understood. We found that increased IER2 expression in human melanoma is associated with shorter overall survival, and subsequently investigated the mechanisms through which IER2 exerts this effect. In experimental melanoma models, sustained expression of IER2 induced senescence in a subset of melanoma cells in a p53/MAPK/AKT-dependent manner. The senescent cells produced a characteristic secretome that included high levels of the extracellular phosphoglycoprotein osteopontin. Nuclear localization of the IER2 protein was critical for both the induction of senescence and osteopontin secretion. Osteopontin secreted by IER2-expressing senescent cells strongly stimulated the migration and invasion of non-senescent melanoma cells. Consistently, we observed coordinate expression of IER2, p53/p21, and osteopontin in primary human melanomas and metastases, highlighting the pathophysiological relevance of IER2-mediated senescence in melanoma progression. Together, our study reveals that sustained IER2 expression drives melanoma invasion and progression through stimulating osteopontin secretion via the stochastic induction of senescence

European small pelagic fish distribution under global change scenarios

The spectre of increasing impacts on exploited fish stocks in consequence of warmer climate conditions has become a major concern over the last decades. It is now imperative to improve the way we project the effects of future climate warming on fisheries. While estimating future climate-induced changes in fish distribution is an important contribution to sustainable resource management, the impacts on European small pelagic fish—representing over 50% of the landings in the Mediterranean and Black Sea between 2000 and 2013—are yet largely understudied. Here, we investigated potential changes in the spatial distribution of seven of the most harvested small pelagic fish species in Europe under several climate change scenarios over the 21st century. For each species, we considered eight Species Distribution Models (SDMs), five General Circulation Models (GCMs) and three emission scenarios (the IPCC Representative Concentration Pathways; RCPs). Under all scenarios, our results revealed that the environmental suitability for most of the seven species may strongly decrease in the Mediterranean and western North Sea while increasing in the Black and Baltic Seas. This potential northward range expansion of species is supported by a strong convergence among projections and a low variability between RCPs. Under the most pessimistic scenario (RCP8.5), climate-related local extinctions were expected in the south-eastern Mediterranean basin. Our results highlight that a multi-SDM, multi-GCM, multi-RCP approach is needed to produce more robust ecological scenarios of changes in exploited fish stocks in order to better anticipate the economic and social consequences of global climate change

Hyporheic Zone Microbiome Assembly Is Linked to Dynamic Water Mixing Patterns in Snowmelt‐Dominated Headwater Catchments

Terrestrial and aquatic elemental cycles are tightly linked in upland fluvial networks. Biotic and abiotic mineral weathering, microbially mediated degradation of organic matter, and anthropogenic influences all result in the movement of solutes (e.g., carbon, metals, and nutrients) through these catchments, with implications for downstream water quality. Within the river channel, the region of hyporheic mixing represents a hot spot of microbial activity, exerting significant control over solute cycling. To investigate how snowmelt‐driven seasonal changes in river discharge affect microbial community assembly and carbon biogeochemistry, depth‐resolved pore water samples were recovered from multiple locations around a representative meander on the East River near Crested Butte, CO, USA. Vertical temperature sensor arrays were also installed in the streambed to enable seepage flux estimates. Snowmelt‐driven high river discharge led to an expanding zone of vertical hyporheic mixing and introduced dissolved oxygen into the streambed that stimulated aerobic microbial respiration. These physicochemical processes contributed to microbial communities undergoing homogenizing selection, in contrast to other ecosystems where lower permeability may limit the extent of mixing. Conversely, lower river discharge conditions led to a greater influence of upwelling groundwater within the streambed and a decrease in microbial respiration rates. Associated with these processes, microbial communities throughout the streambed exhibited increasing dissimilarity between each other, suggesting that the earlier onset of snowmelt and longer periods of base flow may lead to changes in the composition (and associated function) of streambed microbiomes, with consequent implications for the processing and export of solutes from upland catchments.Geological Society of America; Central Ohio Gem and Mineral Society; OSU's School of Earth Sciences' Friends of Orton Hall; Biological and Environmental Research program in the U.S. DOE Office of Science [DE-SC0016488]; Watershed Function Scientific Focus Area at Lawrence Berkeley National Laboratory; U.S. Department of Energy (DOE) Subsurface Biogeochemical Research Program, DOE Office of Science, Office of Biological and Environmental ResearchUnited States Department of Energy (DOE) [DE-AC0205CH11231]; Office of Biological and Environmental Research (BER)6 month embargo; published online: 26 October 2019This item from the UA Faculty Publications collection is made available by the University of Arizona with support from the University of Arizona Libraries. If you have questions, please contact us at [email protected]