EndocRinE disruptors throughout life: interferenCes with develOpmental, reproductiVE and cardiovasculaR health
Progetto The RECOVER project will explore molecular mechanisms underlying the effects of endocrine disruptors (EDs) plastic pollutants that contaminate the environment, including Bisphenols (BPs) and perfluoroalkyles (PFs), in promoting health disorders mainly occurring at different stages of life, from alterations of fetal development to infertility and cardio-vascular diseases. To this purpose experimental research protocols will be carried out to assess: a) molecular mechanisms of oxidative and apoptotic damage induced by BPA in human spermatozoa, b) the biological effects of BPs/PFs on human endothelial cells and c) the epigenetic and molecular mechanisms by which the in utero exposure to BPs/PFs can alter the integrity of the stem cells and affect human development. The hypothesis of the existence of common biochemical molecular pathways through which EDs can exert their biological effects in different cells will be tested. Specifically, it will be investigated the signalling leading to the activation of the enzyme ABHD2 which hydrolyzes the endocannabinoid 2-AG modulating calcium fluxes within the cell. This pathway, partly characterized in human sperm could be active also in other cells, given the widespread expression of the ABHD2. Advanced cytofluorometric techniques will be used to study the apoptotic/oxidative damage induced by EDs on human spermatozoa. The molecular analysis of the proatherogenic effects of EDs will be carried out using endothelial cells in both classical culture systems and a microfluidic system, a model that closely mimics the physiological fluidic shear stress. Biological, biochemical, transcriptomic, and epigenomic effects of EDs on human fetal development will be assessed combining data from human induced pluripotent stem cells and perinatal stem cells in an innovative biological model that mimics the prenatal condition. Biochemical analyses will include the assessment of ABHD2 hydrolytic activity and the quantification of intracellular levels of 2-AG by liquid chromatography-atmospheric pressure chemical ionization-mass spectrometry, both in the presence and absence of EDs.
Unravelling the molecular mechanisms through which BPs and PFs negatively interfere with the unctional integrity of human sperm as well as endothelial and stem cells might open new strategies in preventing their impact on male fertility, cardiovascular and development disorders. In particular, results from the RECOVER project could prompt preventive approaches based on the use of new molecules targeting the ABHD2/2-AG pathway. Identification of damage mechanisms may also be relevant to set up new diagnostic tools to monitor cellular health conditions in response to different degrees of exposure to EDs. Finally, results from dose-response experiments could help the regulatory agencies to set appropriate tolerable daily intakes, thus reducing human exposure and environmental pollution by BPs/PFs.