Anagrafe della ricerca

Ischemic heart disease is the major cause of mortality worldwide, making the demand for safe and effective cardiac regeneration approaches an urgent need. In vitro pathological cardiac tissue models may support efficient design and preclinical validation of new advanced therapies in compliance with 3Rs principle (Reduction, Refinement, Replacement). Among investigated approaches, non-viral direct reprogramming (DR) of cardiac fibroblasts (CFs) into induced cardiomyocytes (iCMs) triggered by microRNA release has emerged as a promising strategy for in situ treatment and cardiac function recovery, although it is limited by low efficiency. DESIRE project aims at exploiting interdisciplinary and complementary expertise, including tissue engineering, biomaterial design, nanomedicine, human stem cell extracellular vesicle biology, microfluidics and in vitro modelling, to significantly enhance cardiac DR efficiency. DESIRE will design an advanced DR approach for post-infarct cardiac regeneration based on in situ delivery of four reprogramming microRNAs, called miRcombo. To improve treatment specificity, efficiency and safety, miRcombo-loaded lipoplexes will be surface decorated with hyaluronic acid (HA-LPs), to bind CD44 receptors overexpressed by CFs during post-infarct inflammation. To improve in situ retention of HA-LPs/miRNA, they will be administered by an injectable bioactive hydrogel, obtained by blending natural polymers, such alginate (ALG), its derivative di-aldehyde alginate (ADA) and gelatin modified with carbohydrazide (GEL-C), forming a double crosslinked network by calcium ions and Shiff’s base reaction. Human amniotic fluid stem cell-derived extracellular vesicles (hAFS-EVs) providing paracrine cardio-protection, antifibrotic and anti-inflammatory signaling will be co-encapsulated in the hydrogel for gradual release to further enhance DR efficiency (“priming treatment”). The bioactive hydrogel will be provided with proper biochemical and biophysical cues to maximize DR efficiency and iCM maturation, and will be validated in a novel human cardiac scar on a chip simulating post-injection hydrogel-tissue interaction. The model will also impart in vivo-like mechanical stimulation, responsible for the establishment of a fibrotic microenvironment and will allow live-recording of electrophysiological parameters. In vitro preclinical validation of DESIRE advanced approach will involve a comparison of the structural and functional properties of iCMs with human adult CMs and induced pluripotent stem cell-derived CMs, healthy cardiac tissue model (positive control) and untreated cardiac scar model (negative control). In vitro preclinically-validated advanced approach from DESIRE will support new technological developments and collaborative projects at EU level for progressive translation into clinical settings. Perspective applications include cardiac regeneration from ischemic, acquired or inherited heart fibrotic disease.