Global gene expression analysis of canine osteosarcoma stem cells reveals a novel role for COX-2 in tumour initiation

Osteosarcoma is the most common primary bone tumour of both children and dogs. It is an aggressive tumour in both species with a rapid clinical course leading ultimately to metastasis. In dogs and children distant metastasis occurs in >80% of individuals treated by surgery alone. Both canine and human osteosarcoma has been shown to contain a sub-population of cancer stem cells (CSCs), which may drive tumour growth, recurrence and metastasis, suggesting that naturally occurring canine osteosarcoma could act as a preclinical model for the human disease. Here we report the successful isolation of CSCs from primary canine osteosarcoma, as well as established cell lines. We show that these cells can form tumourspheres, and demonstrate relative resistance to chemotherapy. We demonstrate similar results for the human osteosarcma cell lines, U2OS and SAOS2. Utilizing the Affymetrix canine microarray, we are able to definitively show that there are significant differences in global gene expression profiles of isolated osteosarcoma stem cells and the daughter adherent cells. We identified 13,221 significant differences (p = 0.05), and significantly, COX-2 was expressed 141-fold more in CSC spheres than daughter adherent cells. To study the role of COX-2 expression in CSCs we utilized the COX-2 inhibitors meloxicam and mavacoxib. We found that COX-2 inhibition had no effect on CSC growth, or resistance to chemotherapy. However inhibition of COX-2 in daughter cells prevented sphere formation, indicating a potential significant role for COX-2 in tumour initiation

Guidelines for the use and interpretation of assays for monitoring autophagy (4th edition)1.

In 2008, we published the first set of guidelines for standardizing research in autophagy. Since then, this topic has received increasing attention, and many scientists have entered the field. Our knowledge base and relevant new technologies have also been expanding. Thus, it is important to formulate on a regular basis updated guidelines for monitoring autophagy in different organisms. Despite numerous reviews, there continues to be confusion regarding acceptable methods to evaluate autophagy, especially in multicellular eukaryotes. Here, we present a set of guidelines for investigators to select and interpret methods to examine autophagy and related processes, and for reviewers to provide realistic and reasonable critiques of reports that are focused on these processes. These guidelines are not meant to be a dogmatic set of rules, because the appropriateness of any assay largely depends on the question being asked and the system being used. Moreover, no individual assay is perfect for every situation, calling for the use of multiple techniques to properly monitor autophagy in each experimental setting. Finally, several core components of the autophagy machinery have been implicated in distinct autophagic processes (canonical and noncanonical autophagy), implying that genetic approaches to block autophagy should rely on targeting two or more autophagy-related genes that ideally participate in distinct steps of the pathway. Along similar lines, because multiple proteins involved in autophagy also regulate other cellular pathways including apoptosis, not all of them can be used as a specific marker for bona fide autophagic responses. Here, we critically discuss current methods of assessing autophagy and the information they can, or cannot, provide. Our ultimate goal is to encourage intellectual and technical innovation in the field

A good life? a good death? reconciling care and harm in animal research

Laboratory animal science represents a challenging and controversial form of human-animal relations because its practice involves the deliberate and inadvertent harming and killing of animals. Consequently, animal research has formed the focus of intense ethical concern and regulation within the UK, in order to minimize the suffering and pain experienced by those animals whose living bodies model human diseases amongst other things. This paper draws on longitudinal ethnographic research and in-depth interviews undertaken with junior laboratory animal technicians (ATs) in UK universities between 2013 and 2015, plus insights from interviews with key stakeholders in laboratory animal welfare. In our analysis, we examine four key dimensions of care work in laboratory animal research: (i) the specific skills and sensitivities required; (ii) the role of previous experiences of animal care; (iii) the influence of institutional and affective environments and (iv) experiences of killing. We propose that different notions of care are enacted alongside, not only permitted levels of harm inflicted on research animals following research protocols, but also harms to ATs in the processes of caring and killing animals. Concluding, we argue for greater articulation of the coexistence of care and harms across debates in geography about care and human-animal relations

Unsettling antibiosis: how might interdisciplinary researchers generate a feeling for the microbiome and to what effect?

Decades of active public health messaging about the dangers of pathogenic microbes has led to a Western society dominated by an antibiotic worldview; however recent scientific and social interest in the microbiome suggests an emerging counter-current of more probiotic sentiments. Such stirrings are supported by cultural curiosity around the ‘hygiene hypothesis’, or the idea it is possible to be ‘too clean’ and a certain amount of microbial exposure is essential for health. These trends resonate with the ways in which scientists too have adopted a more ‘ecological’ perspective on the microbiome. Advances in sequencing technologies and decreasing costs have allowed researchers to more rapidly explore the abundance and diversity of microbial life. This paper seeks to expand on such probiotic tendencies by proposing an interdisciplinary methodology researchers might use to generate more-than-antibiotic relations between lay participants and their domestic microbiome. The paper draws on findings from an ESRC-funded study, Good Germs, Bad Germs: Mapping microbial life in the kitchen (grant number ES/N006968/1), which sought to: (i) explore human-microbe relations in the domestic kitchen; and (ii) make scientific techniques for visualising the domestic microbiome available to non-expert publics through a form of ‘participatory metagenomics’. We examine how scientific knowledge and techniques are enroled into lay practices of making microbes sensible; how these intersect with, reinforce or disrupt previous feelings for microorganisms; and how new ways of relating with microbial others emerge. In reflecting on these findings we draw on work in animal geographies, environmental humanities and the social science of the microbiome. We examine how cultural, emotional and embodied responses to nonhuman others—their ability to affect ‘us’ humans—have implications for the ways in which public health authorities, researchers and ‘lay’ publics alike seek to engage with and govern nonhuman life. We argue that understanding and potentially generating different modes of relating to microbes—a feeling for the microbiome—offers opportunities for reconfiguring microbiopolitics and intervening into the ways in which publics respond to perceived microbial opportunities and threats

Developing a Collaborative Agenda for Humanities and Social Scientific Research on Laboratory Animal Science and Welfare

Improving laboratory animal science and welfare requires both new scientific research and insights from research in the humanities and social sciences. Whilst scientific research provides evidence to replace, reduce and refine procedures involving laboratory animals (the ‘3Rs’), work in the humanities and social sciences can help understand the social, economic and cultural processes that enhance or impede humane ways of knowing and working with laboratory animals. However, communication across these disciplinary perspectives is currently limited, and they design research programmes, generate results, engage users, and seek to influence policy in different ways. To facilitate dialogue and future research at this interface, we convened an interdisciplinary group of 45 life scientists, social scientists, humanities scholars, non-governmental organisations and policy-makers to generate a collaborative research agenda. This drew on methods employed by other agenda-setting exercises in science policy, using a collaborative and deliberative approach for the identification of research priorities. Participants were recruited from across the community, invited to submit research questions and vote on their priorities. They then met at an interactive workshop in the UK, discussed all 136 questions submitted, and collectively defined the 30 most important issues for the group. The output is a collaborative future agenda for research in the humanities and social sciences on laboratory animal science and welfare. The questions indicate a demand for new research in the humanities and social sciences to inform emerging discussions and priorities on the governance and practice of laboratory animal research, including on issues around: international harmonisation, openness and public engagement, ‘cultures of care’, harm-benefit analysis and the future of the 3Rs. The process outlined below underlines the value of interdisciplinary exchange for improving communication across different research cultures and identifies ways of enhancing the effectiveness of future research at the interface between the humanities, social sciences, science and science policy