Search

The project focuses on the modulation of fibrotic phenotype up-set in rare diseases through a Toll-like Receptor 4 (TLR4) antagonist. In particular, in this work, we started to test the possible impact on fibrosis progression of FP7 and FP12 that showed inhibition activity against TRL4 activation by LPS in a dose-dependent way in both human and murine cells 1. Fibrosis is an outcome of the repair response to tissue damage caused by inflammation. When the fibrotic process is excessive or dysregulated it leads to a pathological condition that can affect different organs and functions. Here, it is now clear that inflammation, which however is not the only trigger, plays a key role in the critical cellular process of fibroblast activation that leads to fibrosis up-set 2. The recent discovery of complex crosstalk between fibrosis progression and inflammatory pathways underlines the central role of TRL4 and its potential as a new drug target3. Here it is proposed an in vitro screening on cellular models of fibrosis with TLR4 antagonists to identify new potential drugs targeting Idiopathic Pulmonary Fibrosis (IPF), a rare fibrotic pathology where a pivotal role of TLR4-mediated inflammation has been observed 4 5.

In this work, we present the potential of Fourier transform infrared (FTIR) microspectroscopy to compare on whole cells, in an unbiased and untargeted way, the capacity of bacterial lipopolysaccharide (LPS) and two rationally designed molecules (FP20 and FP20Rha) to activate molecular circuits of innate immunity. These compounds are important drug hits in the development of vaccine adjuvants and tumor immunotherapeutics. The biological assays indicated that FP20Rha was more potent than FP20 in inducing cytokine production in cells and in stimulating IgG antibody production post-vaccination in mice. Accordingly, the overall significant IR spectral changes induced by the treatment with LPS and FP20Rha were similar, lipids and glycans signals being the most diagnostic, while the effect of the less potent molecule FP20 on cells resulted to be closer to control untreated cells. We propose here the use of FTIR spectroscopy supported by artificial intelligence (AI) to achieve a more holistic understanding of the cell response to new drug candidates while screening them in cells.

We present here preliminary data on a study on the possible impact of chemical modulation of TLR4 activity on fibrosis progression. Our labs developed several synthetic molecules that showed biological activity in inhibiting TRL4 activation by LPS in a dose-dependent way in both human and murine cells. Our in vitro experiments with purified receptors suggested that the antagonistic action is due to the interaction of these compounds with MD-2 and CD14 co- receptors (1). Fibrosis is an outcome of the repair response to tissue damage caused by inflammation. When the fibrotic process is excessive or dysregulated it leads to a pathological condition that can affect different organs and functions. Here, is now clear that inflammation, which however is not the only trigger, plays a key role in the critical cellular process of fibroblasts activation that leads to fibrosis upset (2). The recent discovery of a complex crosstalk between fibrosis progression and inflammatory pathways suggests the central role of TRL4 and its potential as new drug target(3). Here we report an in vitro screening on cellular models of fibrosis of a variety of synthetic TLR4 antagonists to identify new potential drugs targeting fibrotic diseases. Thus, the aim is to identify new or old compounds acting on TR4 to block or prevent the fibrosis development. Principal target disses are Idiopathic Pulmonary Fibrosis (IPF) and Morphea which are rare fibrotic pathologies where a pivotal role of TLR4-mediated inflammation has been observed (4) (5). References: 1. Facchini FA, Zaffaroni L, Minotti A, Rapisarda S, Calabrese V, Forcella M, et al. Structure– Activity Relationship in Monosaccharide-Based Toll-Like Receptor 4 (TLR4) Antagonists. J Med Chem. 12 aprile 2018;61(7):2895–909. 2. Upcoming treatments for morphea - Wenzel - 2021 - Immunity, Inflammation and Disease - Wiley Online Library [Internet]. [citato 11 maggio 2023]. Disponibile su: https://onlinelibrary.wiley.com/doi/full/10.1002/iid3.475. 3. B

Bioimaging has drastically transformed the field of medicine, and made the process of diagnosis easy and fast. Visualization of complete organ to complex biological processes has now become possible. Among the various imaging processes, fluorescence imaging using nontoxic fluorescent nanomaterials is advantageous for several beneficial features including high sensitivity, minimal invasiveness, and safe detection limit. In this study, we have synthesized and characterized a new class of nontoxic, self-activated fluorescent hydroxyapatite nanoparticles (fHAps) with different aspect ratios (thin-rods, short-rods, rods) by changing the stabilizing agents (triethyl amine and acetyl acetone) and solvents (water and dimethyl sulfoxide). fHAps showed excellent fluorescence with a broad emission spectrum ranging from 350 to 750 nm and maximum at 502 nm. The presence of fluorescence was attributed to the electronic transition in the asymmetric structure of fHAps as confirmed by ESR spectroscopy and the absence of fluorescence in symmetric HAp NPs. In addition to exceptional fluorescence behavior, these NPs were found to be nontoxic in nature and could be easily internalized in both prokaryotic and eukaryotic systems. We propose that the fHAps provide a safe and a potential alternative to the current fluorescent materials in use for biolabeling and bioimaging applications.