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.

Human populations are impregnated by various types of chemical pollutants reported as “hazardous waste” present in the environment. Certain pollutants have adverse effects on brain, but the underlying mechanisms and time-window exposures are still unknown, as well as their contribution on dementia, the 7th leading cause of death. EXPOSIGNALZ project aims to delineate the impact of a selection of environmental pollutants on brain health throughout life and their role in dementia especially Alzheimer’s disease (AD) representing about 70% of dementia cases. Through interdisciplinary approaches, integrating experimental and epidemiological studies, our objectives are to: 1) Identify environmental pollutants likely to have neurotoxic effects and pro-amyloidogenic properties predictive of a neurodegenerative trajectory related to AD using in vitro screening models; 2) Characterize pollutant signatures associated with brain aging and AD in biological matrices of 4 European population-based cohorts of various age groups; 3) Understand the mechanisms of action of pollutants identified in the in vitro screenings and those found in biological signatures, using AD-iPSC and AD preclinical models; 4) Explore the impact of pollutants on early neurodevelopment as a factor of susceptibility for later neurodegenerative diseases using brain organoids and gestational contamination models and 5) Disseminate knowledge to policy makers, to the relevant stakeholders and general public in order to define guidelines for disease prevention and take actions for population’s health. The project, which aims to identify chemical pollutants as new risk factors for dementia as well as new pollutant-associated biomarkers, could contribute to: (i) reduce or delay the incidence of AD, and thus reduce the economic and social burden; and (ii) allow an earlier diagnosis of AD combined with new disease modifying treatments to delay the entrance of patients in the more severe stages of the pathology.