The main scientific objective of SECRET is to advance translational research by focusing on the regenerative and therapeutic potential of human perinatal cells, including MSC derived from the amniotic membrane (hAMSC), umbilical cord (hUC/WJ-MSC), and amniotic fluid (hAFSC). The project will thoroughly characterize the cellular secretome (conditioned medium as a whole, CM or Evs concentrated thereof) by integrating cell biology, molecular analysis, and proteomics. The project will develop innovative disease models for myocardial infarction (MI), ischemic stroke (IS), and multiple sclerosis (MS) to explore perinatal cell therapies in conditions with unmet clinical needs. Optimal delivery strategies will be identified using biomaterials tailored to the secretome and the targeted organs, while fostering collaboration through secondments and training.

The research program has thus been structured into 3 scientific work-packages (WP 2-3-4) to reflect its key areas of innovation, from in vitro mechanism of action to delivery systems development and finally in vivo validation for clinical applications. Whereas WP1, 5 and 6 will assist network-wide training initiatives, data administration, project management, DoC recruiting, and consortium communications. 

Projects of each DC will run independently; however, the stand-alone projects are designed to complement each other and, through synergies, identify optimal, new products for the effective treatment of MI, IS and MS patients.

WP2: Cells and SECRET fractions, Lead Prof. Ornella Parolini,  Università Cattolica del Sacro Cuore, Italy

WP2 aims to optimize pre-conditioning strategies for bioactive secretome formulations from hAMSC, hUC/WJ-MSC, and hAFSC, and optimize EV production strategy. The study will harmonize analysis on paracrine potential of PC secretomes by comparing them from three sources simultaneously. Perinatal cell-derived CM/EVs, with or without pre-conditioning, will be characterized for their differential paracrine molecular signature by RNA-seq and proteomic profiling analyses, as well as for their functional phenotype. Clonal cell lines for secretome and EV production will be generated and cultured under GMP-transferable conditions. Techniques include advanced multi-color flow cytometry, mass spectrometry, RNAseq, 3D microfluidic platforms accommodating phenotypic fibrosis and angiogenic assays. 

WP3: SECRET mechanisms of action, Lead Prof. Sveva Bollini, Università degli Studi di Genova, Italy

WP3 aims to determine the optimal human perinatal cell secretome for cardiac repair, neuro-protection and neurological recovery in the context of MS and IS. Experiments will be conducted on mouse and human advanced pre-clinical in vitro and in vivo models. In particular validation of the therapeutic efficacy of perinatal derivatives will be performed on i) iPSC-derived cardiomyocytes, ii) human cardiac organ-on-a-chip system, iii) iPSC-derived brain organoids, iv) mouse models of MI, MS, and IS.

WP4: SECRET delivery methods, Lead Dr Ricardo Pires, Universidade Do Minho, Portugal

The study aims to develop supramolecular hydrogels resembling the cardiac/brain extracellular environment. Hydrogel and nanocarrier formulations will be optimized for encapsulation and controlled release of secretome fractions, and suitability for targeted delivery to the brain and to the heart. The systems will be validated in in vitro and in vivo pre-clinical models.