G protein-coupled receptors (GPCR) are very good targets for drugs. Their presence in the cell membrane make them accessible to drugs, and these receptors are involved in the vast majority of cellular processes. Indeed, GPCR are targeted by more than 30% of marketed drugs. To increase the efficacy and decrease adverse side effects of these drugs, a better comprehension of GPCR signalling is necessary. Knowledge concerning the different receptors has drastically increased these last years. The downside of this phenomenon is the profusion of omics data and scientific papers, the integration of which is a real challenge. The objective of ABLISS is the development of a method for building these signalling networks from available data as a whole: literature and large-scale datasets. The method will encompass two main components: (1) a natural language processing module, allowing to extract and format experimental results from scientific papers and (2) a knowledge-based method, allowing the inference of the network from these results. The framework will be applied to the deciphering of GPCR-triggered ß-arrestin- and ERK-dependent signalling. A first workpackage will be devoted to the knowledge-based method. The principle will be the formalization in ASP (Answer Set Programming) of the reasoning that allows the expert deducing network elements from experimental results. We have developed a first prototype, and thus demonstrated the feasibility of our approach. In ABLISS, we will extend the rules and predicate database to cover more experiment types, but also to adapt the reasoning module to the predicate-arguments structures that can be automatically generated by the natural language processing module. We will also study the reliability of a deduced fact. Finally, we will develop abductive reasoning to propose experimental plan allowing verifying hypotheses within the network. A second workpackage will concern the natural language processing module. During the preliminary work on knowledge-based network inference, the necessary manual extraction and formalization of experimental facts has appeared as a major limitation. We have shown, for a limited number of experimental results, that a transducer cascade allows extracting and formatting predicate-arguments structures directly from scientific publications. In ABLISS we will pursue this task, in particular through the development of a transducer cascade allowing extraction and formalization of experimental facts obtained through a large diversity of experiment types. Iteratively, we will ensure the completeness of this predicate ensemble. Finally, we will develop modules to complete the arguments of a predicate when these are not all present locally. The third workpackage will apply the framework to the building of ERK- and ß-arrestin-dependent signalling triggered by different GPCR. A first reason for this choice is that the concerned scientific publications corpus is relatively modest (around 1300 publications), allowing a manual control of obtained results. A second reason is the expertise of the partner coordinating the project in this particular area. New knowledge hypothesized in the network will be validated experimentally.

Growing evidence demonstrates that emotional communication takes place between humans and domestic animals, but most studies have focused on the visual and acoustic channels, neglecting olfactory communication, the most primitive and widespread channel. Recent studies of our teams show that equids and bovidae can perceive human olfactory signals associated with different emotional states, and these chemical cues have begun to be characterized. These first results lead us to investigate further the chemical communication of emotions from humans to animals but also from animals to humans. We propose a project on two domestic livestock species, equids (Equus caballus) and bovidae (Ovis aries), with three aims: (1) To determine whether these species discriminate between different human emotional odours and whether these odours induce emotions in animals; (2) To investigate reciprocity, namely are humans able to discriminate animal emotional odours and how do they react emotionally to these odors? (3) To analyze the chemical composition of emotional odours produced by humans, horses and sheep to confirm whether chemical differences exist between odours from different emotional states in a species, and determine whether the three species share chemical signatures linked to emotional states. Regarding methods, collection of human emotional odours will be based on well-established and published protocols. The participants will watch emotion-charged film extracts (displaying fear, joy, sadness or inspiring disgust) while equipped with under-arm gauze pads. Animal emotional odours will be collected following the same principle. Horses and sheep will experience situations of contrasting emotional valence (positive, such as pleasant touch contact, and more negative, such as a novel environment), while equipped with gauze pads placed under abdominal belts. These odours will then be tested on receiver subjects in appropriate tests for each species. Thus, horses and sheep will participate in ‘habituation/ discrimination’ tests which will enable us to demonstrate whether they can discriminate different human emotional odours. They will then undergo a battery of tests to evaluate their emotional reactivity (e.g., a neophobia test) according to whether they are exposed or not to human emotional odours. The human subjects will take part in specific tests to determine whether or not they can evaluate implicitly or explicitly the animal odours (e.g., standardized laboratory tests in which they will self-evaluate their emotional state before and after smelling an odour). The consortium is well-experienced in the battery of tests the animal and human participants will perform. Finally, to describe the chemical composition of the emotional signals in human and animal sweat, the organic volatile compound extracts will be analyzsed through gas chromatography -mass spectrometry. This inter disciplinary project will group fields such as cognitive ethology, human psychology and chemical ecology. It should improve understanding of how animals and humans mutually communicate their emotions through chemicals. Four different scientific teams will collaborate on the project, which will be supported by scientists recognized in the domains of animal cognition, emotions and welfare, the human-animal relationship, olfactory processing in animals and humans, and chemical ecology.

KISS project associates increasing livestock productivity and sustainability with decreased health risk. Our ambition is to develop a new treatment for reproduction control improving reproductive performance and reducing hormone use. This new treatment will be based on the development of analogs of the endogenous neuropeptide kisspeptin (Kp). A key objective of farming sustainable intensification is improvement of reproduction control in livestock. Our goal is to develop a new treatment to increase reproductive success and to obtain a superior control of birth timing while reducing hormone use and working cost. Current treatments face 4 major problems: i) use of hormone which could persist in the food and in the environment (e.g. progesterone derivatives). If more restrictive laws banning hormonal treatment will be approved, farmers will be obliged to apply less efficient and more time consuming methods leading to income reduction. This could render economically untenable the management of small farm and results in a dramatic drop of their number causing a deconstruction of local social tissue. ii) sanitary risk due to potential diseases transmission (i.e. use of equine Chorionic Gonadotropin (eCG), also named PMSG for pregnant mare’s serum gonadotrophin, extracted from blood serum), iii) immunological response against eCG that affects efficacy of ensuing treatments, and iv) suboptimal efficiency. To address these problems we designed a 1st generation of Kp analogs. Tests in sheep showed that these analogs are active at extremely low doses (tens of µg). Consequently any potential food contaminant issued from Kp analogs would be present in very low amount. In addition, thanks to their peptide nature, Kp analogs will be timely degraded and their inactive building block (simple amino acids) would be rapidly recycled or excreted. Furthermore, preliminary experiments in ewe showed that our 1st generation Kp analogs are superior to eCG in synchronizing ovulation and suggest that they may also improve efficiency. These results imply that replacement of eCG by Kp analogs is foreseeable. This would be by itself a major improvement eliminating sanitary risks and immunogenic liability. To complete the proof of concept tests to induce ovulation in sheep during the non breeding season are planned. Nonetheless, to assure an optimal response we anticipate the need to improve pharmacokinetics and pharmacodynamics of our 1st generation analogs. Therefore, we will develop a 2nd generation of molecules with refined pharmacokinetics and pharmacodynamics optimally suited for management of livestock reproduction. On the other hand, relevant features of Kp system, that may have an impact on Kp analogs efficacy, have been poorly investigated (e.g. receptor desensitization mechanisms, existence of functionally selective agonists, precise distribution of Kp receptor, etc). The lack of a sufficient portfolio of Kp receptor ligands with varied chemical structures and pharmacological profiles (partial agonists, antagonists, and biased agonists) largely account for the paucity of information on these topics. Our chemistry effort is generating a series of analogs with different chemical structures, highly selective and with distinct pharmacological profiles. We have the unique opportunity to exploit these new pharmacological tools, and eventually modify them to create antagonists, to better understand Kp system properties. This information will represent an important advance in basic science and will help the design of optimized analogs. Finally an added value of Kp analogs could be their use as a first step to develop new therapeutic agents to treat human diseases. To summarize, the objectives of the present project are 1) to develop a 2nd generation of Kp analogs 2) to ameliorate livestock reproduction management and 3) to advance our understanding of Kp system physiological functions.

The control of female reproduction has become a major societal and economic concern, and thus a better understanding of the central mechanisms acting on the reproductive axis is necessary. Early studies on the neural pathways involved in the control of the reproductive axis have highlighted the pivotal role of GnRH (Gonadotropin Releasing Hormone) neurons located in the rostral hypothalamus. GnRH release into the portal blood stimulates the secretion of the pituitary gonadotropins which are critical to trigger puberty and to regulate reproductive fonction. In recent years, however, studies have revealed that hypothalamic neurons producing peptides of the RF-amide (Arg-Phe-NH2) family, especially kisspeptins (Kp) and RFamide-related peptide-3 (RFRP-3), play a key role in the control of reproduction. These neuropeptides appear to act upstream of GnRH neurons, and current findings in this domain are leading to a new model for the neuroendocrine control of reproduction. Kp neurons, located in the arcuate and anteroventral periventricular nuclei, project to GnRH cell bodies and nerve terminals. There, Kp binds to its specific receptor Kiss1R (or GPR54) to potently stimulate GnRH release. Notably, Kp neurons are the main central targets for the positive and negative feedback effects of sex steroids. Moreover, Kp neurons sense metabolic signals to tune reproductive activity according to the energetic status of the organism. In seasonal breeders, Kp expression is regulated by photoperiod and the peptides synchronise reproduction with seasons. Altogether, current data have demonstrated that Kp is a key regulator of the gonadotropic axis in all mammalian species studied so far. On the other hand, RFRP-3, expressed in neurons of the dorsomedial hypothalamus, was first reported to inhibit reproductive activity by reducing GnRH neuron activity and pituitary gonadotropin release. However, recent data report that RFRP-3 activity depends on species, gender and physiological status, indicating complex mechanisms of action. The distribution and pharmacology of the receptor for RFRP-3, GPR147, are poorly known and the lack of selective pharmacological and genetic tools severely limits the study of the RFRP-3/GPR147system. To complicate the picture, a few studies suggest that GPR147 may bind other endogenous RFamide peptides, including Kp. In this highly competitive context of a renovated understanding of the neuroendocrine control of reproduction in mammals, the main aim of the REPRAMIDE proposal is to elucidate the role and to describe the mechanisms underpinning RFRP-3/GPR147 influence on mammalian reproduction. Our specific objectives are 1) to determine the physiological effect of RFRP-3 on female reproductive activity using three complementary animal models in order to clarify species-specific differences; 2) to generate suitable tools to refine the pharmacological and biochemical knowledge of GPR147; 3) to evaluate the putative interaction between GPR147 and other RF-amide receptors, particularly Kiss1R, both at the pharmacological and physiological levels. The findings obtained in the course of our studies will contribute to the development of better strategies to treat human fertility problem and to manage reproduction in livestock.