Ricerc@Sapienza

Understanding the differences between physiological and pathological autophagy may help us design therapeutic strategies to target pathological autophagy without hindering its physiological effects.
The multidisciplinary approaches involving a team of health professionals from various disciplines will provide combined data, that will elucidate the molecular mechanisms underlying the regulation of autophagy, that is found to be involved in the pathogenesis RA and other autoimmune diseases. These findings will form a basis for future studies into the role of autophagy allowing the discovery of new targets and compounds and the developing of novel types of drugs targeting cellular catabolism process. There are several autophagy modulators in use or under investigation in the management of RA therapy. Moreover, novel therapeutics and interventions are being investigated to counter autophagy-associated diseases, including apoptosis inhibitors and activators. Excessive autophagy stimulation may cause self-destruction which could be controlled with drugs such as vacuolar-type H (+)-ATPase inhibitors, cycloheximide, lysosome alkalizers (chloroquine, hydroxychloroquine, NH4Cl, and neutral red), and acidic protease inhibitors (E64d and pepstatin A), whilst knocking down Beclin 1 and Atg5 using MiR-30a could be used to inhibit the excessive cannibalism caused by autophagy. Upregulating autophagy could therapeutically benefit a range of diseases prevent autoimmune conditions, and also increase lifespan. Rapamycin, small-molecular enhancers of rapamycin (AUTEN-9), trehalose, IMPase inhibitors (carbamazepine and valproic acid), epigenetic modulators (anacardic acid, curcumin, garcinol, and spermidine), and chemicals (fluoxetine, penitrem A, and metformin) are all autophagy stimulators which help to ameliorate disorders associated with reduced autophagy. Syntheses will be performed applying the most modern organic and pharmaceutical chemistry approaches to deliver compounds with maximized purity, yields and throughput. Techniques will include multicomponent reactions, liquid-phase parallel synthesis, microwave and flow chemistry techniques, under pressure and inert reactions, applying selective protective groups and stereoselective synthesis. The structure and purity (>95%) of the molecules will be confirmed by HPLC, LC-MS and NMR spectroscopy. Moreover, we expect to identify novel interactors of the given molecules, their targets and find post-translational modifications turning out to be required in the ontogenesis or progression of other diseases, such as neurodegeneration or cancer. Post-translational modifications encompass a group of reactions that modify the structure and extend the functions of proteins. Two of these modifications, citrullination and carbamylation may play a role in the pathogenesis of RA. Citrullination is an enzymatic post-translational modification that is mediated by PAD, which transform peptide-bound arginine residues into citrulline, a non-natural amino acid. Carbamylation is a chemical post-translational modification consisting of the addition of a cyanate group on self peptides, leading to the production of homocitrulline. In addition, taking advantage of the deeper knowledge of some pathogenic aspects of RA and on the recently discovered cell processes (such as citrullination, autophagy and NETosis) might represent an innovative tool for the discovery of new biomarkers, e.g. characteristics of germs, their enzymatic machinery, their interaction with host immune response and the induction of cellular response affecting survival. Moreover, the understanding of the cell process involved in autoimmune diseases such as NET formation, autophagy, enables physicians to be more confident and more skillful to use drugs for the treatment of autoimmune diseases. The consequences of these expected results could be their application, aiming to evaluate their weight in diagnostic procedures and, even more, prognosis and response to treatment. Our final purpose is to verify the identified biomarkers by larger clinical studies accordingly to specific therapeutic protocols based on the use of new strategies of care (standard DMARDs) or new drugs (biological drugs). All this could have great importance for clinical, preventive and therapeutic aspects, but also great economic relevance (to allow a rationalization of the health care costs). It is important to mention that the new and effective treatments of RA, with the employment of biologic drugs, undergo from the huge costs and the weight of severe side effects. Futhermore, the response of these therapies being sometimes ambiguous and their efficacy is limited in a percentage of patients (about 30%). This scenario imposes the need of discovery new pathways and therapeutic targets. We aim the achievements of the targets of this project will have positive consequences in terms of economic costs as well as for the life quality of patients affected from RA.