The amniotic membrane of human term placenta is a valuable source of cells endowed with several beneficial effects. Amongst these, mesenchymal stromal cells from the amniotic membrane (hAMSCs) have been shown to favour tissue repair and regeneration after transplantation in rodent models of inflammatory-based disease. The benefits of transplanted amniotic cells were observed despite cell engraftment in injured tissue, thus suggesting these cells produce bioactive factors able to mediate these effects through paracrine signaling. Accordingly, beneficial effects in preclinical models, were demonstrated when using cell-free treatments, such as conditioned media containing factors secreted by amniotic cells during their in vitro culture.

Therefore, the mechanisms of action are not based on cell replacement but rather on the repair of damaged tissue by bioactive factors with anti-inflammatory, anti-fibrotic, and anti-angiogenic properties, resulting in the activation of resident stem cells. The results obtained in several animal models are now being exploited in over 20 Phase I and Phase II clinical trials (https://clinicaltrials.gov). It is increasingly more evident that the bioactive factors produced by amniotic cells (i.e. secretome) can favour tissue repair and regeneration through the resolution of inflammation by acting on different inflammatory mediators, however to date, their biochemical nature, as well as the extracellular vesicle fraction(s) in which they are found, have yet to be defined.

Based on this state of the art, the main aims of the project are:

1. to provide a significant advancement in the knowledge of the amniotic secretome; this project will implement high-throughput sequencing and other massively parallel technologies, also known as "omics technologies" (e.g. proteomics, genomics, transcriptomics, lipidomics and metabolomics). Even though each technology individually cannot capture the entire biological complexity, their integration enables a multiparametric/multimodal profiling of molecules and bioactive factors through a system biology approach. This will be performed both on whole secretome and nano/microvesicle fractions;
2. to develop a suitable delivery system, based on the design of innovative hydrogel-based nanofillers entrapping the secretome, functionalized to achieve a controlled and optimized release in the target tissue.
3. to validate the efficacy of the secretome delivered through innovative nanotechnology in a clinically relevant animal model, represented by spontaneous equine osteoarthritis, a paradigm condition in which immune-related mechanisms are prominently involved and lead to irreversible joint damage.

The feasibility of this project is supported by a coordinated, multi-disciplinary group. Indeed, the Italian, multi-disciplinary team in this project is comprised of internationally-renowned researchers including cell and molecular biologists, morphologists, chemists, biochemists, clinical rheumatologists, and veterinary surgeons that have a long-standing experience in the different research and clinical area involved in the project:

• perinatal stem cells including one of the pioneers in this field, the PI of the project;
• development of novel nano-technologies;
• molecular profiling through high-throughput multiomic technologies;
• veterinary medicine related to equine osteoarthritis.

Working group:

• Wanda Lattanzi - Faculty of Medicine and Surgery
• Sabrina Ceccariglia - Facultyof Medicine and Surgery
• Stefano Alivernini - Facultyof Medicine and Surgery
• Tommaso Pirronti - Facultyof Medicine and Surgery

Partners:

• Università Cattolica del Sacro Cuore (Coordinator)
• Università degli Studi di Palermo
• Consiglio Nazionale delle Ricerche
• Università degli Studi di Roma "La Sapienza"
• Università degli Studi di Cagliari
• Università degli Studi "G. d'Annunzio" Chieti-Pescara