Speaker
Description
"Unresolved inflammatory responses in chronic disorders such as diabetes, cancer, cardiovascular diseases, among others, often drive the onset of non-healing wounds. Endothelial cells (EC), the population of cells lining the inner layer of blood vessels, become dysfunctional under the pathophysiological context of chronic disorders and undergo endothelial-to-mesenchymal transition (EndMT), resulting in a fibroblast-like phenotype. During the early stages of wound healing EndMT results in impaired vascularization and the deposition of scar tissue by fibroblasts, which contribute to the onset of non-healing wounds. Targeted therapeutics aimed at reversing EndMT at the site of injury by transdifferentiating fibroblasts to EC have the potential to enhance vascularization, expedite the wound healing process and prevent the chronification of wounds. We developed a collagen membrane functionalized with phosphatidylserine (PS)-containing lipid nanoparticles (LNP) that encapsulate mRNA encoding for proteins inducing fibroblast transdifferentiation into EC (GATA2, ETV2, FLI1, KLF4). Our particles are nanoscopic in size (~70 nm) and can successfully encapsulate over 70% of the RNA loaded into them during their preparation. Furthermore, our LNP are taken up by up to 40% fibroblast populations treated with them, a result that is comparable to other conventional transfection methods such as lipofectamine. Gene expression of the delivered mRNA is also significantly increased in fibroblasts treated with our LNP. For the development of our collagen membranes we devised an approach involving their functionalization with Annexin V, a PS-binding protein, to be used as an anchor to attach our LNP onto the membranes and enhance their retention. Our results also show a successful functionalization of the collagen membranes with Annexin V and our LNP. These membranes have the potential to serve as implants promoting in situ wound healing by providing a niche for resident fibroblasts to transdifferentiate to EC through the uptake of our mRNA-loaded LNP, ultimately enhancing vascularization and expediting tissue repair. The biomimetic approach we propose here for the development of our membranes follows the use of biocompatible materials mimicking the normal composition of the extracellular matrix to avoid the onset of inflammatory responses after their implantation. Moreover, this approach represents a novel alternative to the development of RNA therapeutics to concomitantly encapsulate and deliver ad hoc multiple RNAs, while protecting them from degradation and making it possible for them to exert specific biological functions in target cells. Lastly, the inherent versatility of these mRNA-LNP formulations make them an attractive platform that has the potential for targeting a variety of chronic diseases to improve their treatment and positively impact the quality of life of patients.
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62825451448
