Background Accumulation of extracellular matrix, recruitment of inflammatory macrophages and neutrophils secreting reactive oxygen species (ROS), and cytokines are characteristic of idiopathic pulmonary fibrosis (IPF) and renal fibrosis (RF). Signaling cascades triggered by TGF beta and involving cytokines and profibrotic factors are important therapeutic targets. The pro-inflammatory/pro-angiogenic chemokines ELR+CXCL and their receptors CXCR1/2 play a key role in promoting pulmonary fibrosis and nephropathy. In addition, VEGFC, a key driver of lymphangiogenesis produced by proinflammatory M2 macrophages, is critical for fibrosis via a cross-talk with TGF beta and ELR+CXCL stimulate lymphangiogenesis via VEGFC. To develop small therapies targeting these processes, the coordinator founded start-ups Roca therapeutics (RT) and Kekkan Biologics (KB). RT developed CXCR1/2 inhibitors and its drug candidate RCT001, and KB licensed the coordinator team's anti-VEGFC monoclonal antibodies. RCT001 inhibits the production of ROS, inflammation and fibrosis, is non-toxic to normal cells, stable, has good bioavailability and half-life. Anti-VEGFC Mab inhibits the proliferation of fibroblasts. Hypothesis Our hypothesis stipulates that ELR+CXCL/CXCR1/2 play a crucial role in fibrosis and that targeting CXCR1/2 can stop inflammation, mesenchymal cell activation, angiogenesis, and fibrosis. Interfering with the VEGFC pathway, which is also linked to CXCR1/2 signaling, may be effective in its own right or may further enhance the efficacy of RCT001. We believe that targeting two interconnected signaling pathways involved in chronic inflammation and abnormal lymphangiogenesis may represent innovative therapies. Methodology This translational project is based on the study of patient samples and the correlation between the presence of players in the ELR+CXCL/CXCR1/2 and VEGFC signaling pathways and the severity of IPF and/or RF. The effects of inhibiting both pathways individually or in combination on M2 macrophage polarisation and their ability to inhibit ROS formation will be tested in models of epithelial and endothelial cells and fibroblasts. The relevance of the two targetings will be evaluated functionally or biologically in models of IPF and RF in vivo. Expected results Pulmonary and renal fibrosis are currently at therapeutic impasses. Innovative therapies are urgently needed. Our patented molecules, backed by two start-ups, mean hope for patients. The two start-ups will licence the application patents for these inhibitors to carry out all the necessary steps to enter the clinical phase.

Today, information systems are one of the world's main resources. As accentuated by the Covid-19 crisis, our society relies on an ever-increasing need to process and communicate data, with significant repercussions on politics, defense, health, innovation, daily-life and the economy. The level of security remains a major issue for many use-cases, where secret key encryption are provably secure and can be implemented in the real world via quantum solutions. Quantum-safe communication, the first commercially available quantum technology, provides a unique means to establish, between distant locations, random strings of identical secret bits, with a level of security unattainable using conventional approaches. The implementation of actual quantum systems has become crucial, given the strong military, societal and economic impacts. This path, considered as one of the most promising for IT innovation, benefits from largely endowed R&D programs, such as the EU Flagship and other national initiatives (UK, Germany, China, USA, France). With the development of quantum computers and sensors, it becomes of prime necessity to connect them. Consequently, tasks such as distributed quantum computing and sensing will lead to a large-scale quantum Internet. The major obstacle to the adoption of such networks lies in the limited distance (~100 km) over which they can be deployed, due to losses in optical fibers and the curvature of the Earth. In the absence of reliable quantum repeaters, the space segment represents the only potential way to circumvent this limitation. To date, the only real demonstrations have been made in China (Micius satellite), but many projects are underway at the international scale. SoLuQS aims at effectively answering this demand by building industrial "entanglement source" prototypes that meet the constraints of spatialization, without compromising their performance. The key words of our achievements will be compactness and integrability, allowing satellite exploitation for both civil and military domains. These devices will eventually allow the connection of 2 metropolitan quantum networks (Paris and Nice). SoLuQS will therefore follow the promising path of new telecom-compatible laser optical communication systems in free space, and is thus part of the ASTRID AAP's thematic axis 3, "Cryptography - Communication", with a focus on "network security", their "operational implementation" based on "multimodal entanglement", as well as "space solutions". We will develop, at the French scale, the necessary tools for spatialization, in view of establishing a secure space/ground communication link, in order to anticipate future satellite realizations. SoLuQS brings together the best international teams in quantum communication (INPHYNI and LIP6) as well as a major French space industrial group (Thales Alenia Space) which will promote both integration and spatialization of the achievements. The consortium will pursue an active knowledge dissemination strategy. IP and the attraction of industrialists have a directly exploitable economic value, both in terms of patents, market reach, and creation of start-ups. We will ensure the training of staff and students as well as the promotion of partners in both the academic and industrial communities. These activities will be complemented by dissemination actions (international conferences, scientific and general public publications, etc.) in order to maximize the project impact. Taken as a whole, our actions will ensure France to play a leading role on the international level, in terms of disruptive quantum technologies for space quantum communication.

The superfamily of TGF-ß ligands represents one of the most prominent families of morphogens. These factors have profound effects on many aspects of embryonic development, cell behaviour and homeostasis and malfunction of the pathways associated with these cytokines can lead to a variety of pathologies. Despite intensive research, there are still large gaps in our knowledge regarding the specificity of these ligands, the regulation of the activity of their receptors and the interactions between the TGF-ß pathways and other signalling pathways. In particular, how sources of TGF-ß morphogens are generated and how the resulting morphogen gradients can be translated into patterns of gene expression during early development remain central questions in current developmental biology. This proposal attempts to fill these gaps in our knowledge by characterising novel regulators of dorsal-ventral axis formation upstream and downstream of Nodal and by modelling the gene regulatory network (GRN) activated by this factor. We address this question within the sea urchin model, an organism phylogenetically close to vertebrates but with many advantages for the analysis of regulatory networks in early development. Our first aim is to characterize the early events that shape the Nodal gradient and initiate the downstream GRN that controls dorsal-ventral (D/V) axis formation. Our laboratory has recently discovered several key factors involved in D/V axis formation. We identified a maternal TGF-ß ligand, a transmembrane protein, an ETS domain transcriptional repressor and the JNK kinase as factors critically required to restrict the spatial expression of nodal. The similarity of the phenotypes caused by inactivation of either this maternal TGF-ß ligand, the transmembrane protein or this ETS factor strongly suggests that they act in the same pathway to specify the D/V axis. However, the relationships between these factors and the mechanisms by which they antagonize nodal expression are presently completely enigmatic. To clarify the relationships between these factors we will identify the binding partners of the TGF-ß ligand and perform biochemical analyses and epistasis experiments. In this first aim, we will also determine whether Nodal and/or BMP2/4 work as morphogens, i.e. as long-range, concentration-dependent signalling factors in the sea urchin embryo by using a combination of treatments with recombinant Nodal and BMP2/4 proteins and ectopic expression of mRNAs encoding these ligands or the activated forms of their receptors. The second aim of this project is to extend and model the gene regulatory network activated by Nodal. We recently identified and validated about fifteen novel genes regulated by Nodal encoding a variety of regulatory proteins including transcription factors, cytokines, and secreted proteins most of which have never been characterized. The expression patterns of these genes identify novel regulatory domains and boundaries along both the animal-vegetal and dorsal-ventral axes, revealing an unsuspected complexity in patterning of the ectoderm. We propose to dissect the regulatory mechanisms establishing these new domains, to characterize these novel Nodal target genes and to analyze their function and position in the GRN. Finally, to further test the role of individual components of this network and understand how it achieves both robustness to environmental perturbations such as regulative development, and plasticity to evolutionary scenarios, we will start to construct a logical model of this GRN. Our third and last aim is to start dissecting the mechanisms that allow responding cells to read different levels of Nodal or BMP2/4. We will start to investigate how thresholds of response are encoded in the genome. We will perform detailed bioinformatics analyses on an extended set of Nodal and BMP2/4 target genes to identify and dissect the architecture of the cis-regulatory modules of these selected target genes.

Disorders of sex development (DSD) are very heterogeneous and despite intensive research over the last 30 years, only about 50% of DSDs can be explained on the molecular level. This highlights that our knowledge on mechanisms governing sex determination is still fragmented. In the project SexDiff, we will use mouse and human genetics in combination with transcriptomic analyses and bio-informatics to clarify how sex determination is driven. Sex determination is a developmental process allowing the differentiation of a bipotential precursor in two completely different organs, the testis or the ovary. This decision is driven by the paternal transmission of the Y-linked gene SRY which eventually initiates testicular development by up-regulating the transcription factor SOX9. In absence of SRY, R-spondin1 (RSPO1), an activator of the WNT/?-catenin signalling pathway, initiates ovarian differentiation. Both pathways antagonize each other, and sexual differentiation is determined by the dominant pathway. We have a long-standing interest in mechanisms of mammalian sex determination and have contributed to understanding of function of key genes such as SOX9 and RSPO1 and how these genes promote one sexual fate and repress the other. Despite recent advances our understanding of genetic components and mechanisms of sex determination remain limited. Discovery of novel factors and mechanisms involved in the process is of vital interest as mutations in yet to be discovered genes may be a cause of human reproductive pathologies, particularly DSD. The Wilms’ tumour suppressor WT1 is essential for the differentiation of the gonad. Different variants of WT1 exist and the imbalance of the ratio between the alternative spliced isoforms +KTS/-KTS are the cause of the male-to-female sex reversal in the Frasier syndrome. Recently discovered mutations impacting specifically the 4th DNA binding Zinc finger promote female-to-male sex reversal. This positions WT1 at the crossroad of the cell fate decision during sexual development. In this project we aim to decipher the role of these isoforms/variant in the differentiation of the gonad using mouse models and modified induced pluripotent stem cells (iPSC) and to uncover novel signalling networks regulated by WT1 during gonadal differentiation. We believe that it is important because these newly identified factors may provide a valuable diagnostic tool to understand the aetiology of idiopathic cases of human disorders of sex development. In contrast to testis, much less is known about the mechanisms of ovarian development in mammals. We have recently established that ovarian differentiation is decided before the first signs of sexual differentiation, a concept that breaks with the present view of sex determination. During the course of SexDiff, we will further explore this and characterize the molecular function of Rspo1 and identify new actors of ovarian differentiation using our Rspo1 loss-of-function mutants. These newly identified factors will then be further characterised to establish their causality and contribution to errors of ovarian development in human using the combinatorial approach involving genomics, in-silico, in-vitro, ex-vivo and cellular reprogramming approaches. To achieve these goals, we will employ a whole range of state-of the art techniques including single-cell RNA-Sequencing, cellular reprogramming (using iPSCs) and targeted mutagenesis by CRISPR/Cas9. Using the concerted approaches involving mouse models, human patients, cellular models and in–silico analyses, SexDiff will contribute to an in-depth understanding of the normal and pathogenic development of the gonads. This is envisaged to contribute and enhance our understanding of the aetiology of errors in gonad development in human and pave way for a better diagnosis. SexDiff, therefore, is of interest not only for enhancing our knowledge of fundamental biology but has an implicit clinical application.

--- BACKGROUND --- Light-activated proteins of the rhodopsin family, found in all three domains of life, have also been found to be encoded by viruses, specifically giant viruses. Little is known on these virus-encoded rhodopsins and on their possible role in viral infection. Their structure, their oligomeric organization, and their function in terms of transport and substrates are unknown. In spite of significant amino-acid sequence similarity, we have sufficient preliminary data to show that viral rhodopsins differ greatly from known rhodopsins, and can assemble in complexes resembling human pentameric ligand-gated ion channels, where the agonist is light. These proteins could therefore involve novel biological mechanisms and sustain possible applications in optogenetics. --- OBJECTIVES --- Our aims are to decipher the structure and the function of viral rhodopsins, and to examine the possibility of using these proteins as light-sensitive actuators in mammalian tissues. The results should shed light on the role of viral rhodopsins in host infection and could form the basis of new tools for optogenetic applications. --- CONSORTIUM --- The consortium assembles three partners with demonstrated, complementary expertise in the fields of structural biology (IBS-Membrane, Institut de Biologie Structurale, Grenoble), electrophysiology in model cells (IBS-Channels, Institut de Biologie Structurale, Grenoble), and electrophysiology and optogenetics in mammalian native cells (iBV, Institut de Biologie Valrose, Nice). --- WORKFLOW --- Viral rhodopsins are separated in two phylogenetic groups. We will work on representatives of each group, OLPVRI and OLPVRII. Partner IBS-Membrane will perform structural characterization of OLPVRI and OLPVRII in their different conformational states by X-ray crystallography, time-resolved serial crystallography, and single-particle cryo-electron microscopy using advanced European instruments (Grenoble synchrotron, Hamburg X-ray free electron laser, and Grenoble Titan Krios). The partner will also perform in vitro functional characterization of the proteins. In parallel, partner IBS-Channels will characterize the function the viral rhodopsins in model cells and optimize their properties by protein engineering informed by structural data. Partner iBV will test the optogenetic potential of the proteins in neuronal cells and brain slices. The proposed tasks are extremely challenging. However, the feasibility of the project is asserted by solid preliminary data and proven successful experience of the partners in the corresponding fields of science and methodologies. In particular, a high-resolution structure of OLPVRII in the ground state has already been obtained and functional characterization of OLPVRI has been started.