Alzheimer's disease (AD) affects about 600,000 people in Italy and constitutes one of the most serious problems of social medicine. The few therapies approved so far are poorly effective and only at the initial stages of the disease. It is therefore urgent to find strategies capable of slowing the disease.
Glial activation, increased neuroinflammation and changes in metabolic functions characterize the development of AD. Much interest is focusing on endocannabinoids (ECB), especially for their ability to counteract neuroinflammation.
Aim of the project will be: i) to evaluate the efficacy of FAAH, a pharmacological inhibitor of hydrolase involved in the catabolism of ECB, in restoring autophagy, which is involved in the clearance of the amyloid peptide and is reduced in AD; 2) since we recently discussed the potential of some foods derived from plants to increase the ECB tone, we will evaluate green extracts as a possible source of nutraceuticals to counteract neuroinflammation and enhance autophagy.
The experiments will be carried out in vitro, taking advantage of the murine BV2 microglia cell line, analyzing the pro and anti-inflammatory markers after treatment with amyloid peptide or with proinflammatory factors in the presence of FAAH or plant extracts using immunofluorescence, western blot, real -time PCR. Furthermore, the AD Tg2576 mice will be supplemented with FAAH or with plant extracts. Brain will be analyzed by immunohistochemistry, western blot, real-time PCR for the presence of amyloid plaques and the presence of activated microglia, analyzing its polarization towards a pro- or anti-inflammatory phenotype.
This project will allow to deepen the knowledge on the role of ECB in AD pathogenesis, and the development and production of innovative nutraceutical substances from natural sources and the creation of a controlled supply chain of various plants of food interest to be used as matrices for the production of nutraceuticals and functional foods.
The present project is designed according to an integrated approach in a multidisciplinary team, sharing specific and highly specialized knowledge, which will contribute in obtaining results of excellence in a relatively short time period.
Autophagy is a common phenomenon in eukaryotic cells. It is not only the basic physiological activity of the cells, but also a mechanism of the cells to protect themselves from various types of stressors. Through autophagy, organisms can regulate the balance of protein metabolism, which is essential for the removal of cellular catabolites and the growth of structures and organelles.
Investigations into the biological mechanism of autophagy suggest that insufficient autophagy may be a key factor in promoting the development of neurodegenerative diseases. Our recent results showing the anti-inflammatory role of endocannabinoids in a model of AD in vitro, prompted us to hypothesize that endocannabinoids may improve autophagy, that is severely impaired in AD. This information will deepen the knowledge on pathogenetic mechanisms underlying AD progression as well as suggest new therapeutic paths
This project, in addition to increasing the knowledge on the etiopathogenesis of AD, and on the alteration of autophagic processes during the disease, proposes the development of innovative nutraceuticals based on bioactive molecules from natural sources or typical processes, also with a view to a greater sustainability,
natural extracts and "end products" or processing intermediates of food industry transformations, with the aim of enhancing complex molecular profiles, capable with their multifactorial action of Impact on the development of complex diseases, such as Alzheimer's disease, object of this study. In our recent paper we analyzed the role of some foods in the increase of endocannabinoid tone which plays an important role in the decrease of inflammatory factors that characterize the evolution of the CEO. In our study we will analyze the ability of these plant-derived molecules to enhance the autophagic mechanisms which are decreased in AD and which lead to a decrease in the clearance of amyloid peptides that are deposited in senile plaques, histopathological hallmark of the disease.
This research project aims to use innovative nutraceutical substances from natural sources coming from a controlled supply chain of different plants of food interest to be used as matrices for the production of nutraceuticals and / or functional foods.
During the project, in vitro and in vivo models will be developed for the evaluation of the quality and safety of the new formulations and functional properties of the developed nutraceuticals.
The methods used, also in consideration of the collaboration and synergy with other departments of Sapienza University, will allow young researchers (YR) involved in research to increase their know-how for a better integration into national and international research organizations. Finally, the possibility for YR to work in a multidisciplinary team, will help to improve their knowledge and practice of methodology in the biomedical research area. In addition, considering that at present, a correct understanding of the methods of research will be improved, the YRs will be trained (and followed during the entire project development) to work according to the main issue of the methodology of academic integrity. In this connection it is to remind that the PI prof Rita Businaro completed a course on research integrity, according to the VIRT2UE project, funded by Horizon 2020b research program (grant agreement N 787580). Each PhD student (PhDS) and post-doc YR will be will be supported and followed by the researcher involved in the project and part of their working time will be specifically devoted to the training in the performing a research project according to the international standard guidelines for a correct behaviour in the study plaining, the adequate methodology for experiments setting, the data handling (with particular attention to the reproducibility of the experiments), including special attention to sensitive data (clinical and personal data of subjects enrolled during the study), and finally a correct and ethic production and publication of the whole results of the study. Each YR and PhDs will be involved in all the part of the study organization and particular attention will be reserved to the data handling, in order to directly learn the correct conduct during the progression of the work of eventual project modification in order to attempt at specific problems that were not considered during project ideation
All data obtained with the above mentioned experimental approaches will be published on international scientific journals and presented to national as well as international meetings and congresses.