Currents Fabio Projects

Dengue fever is the most dominant arthropod-borne viral disease, threatening almost half of the world ’s population in 129 countries in tropical and subtropical regions. The efficacy of the only currently approved vaccine (DengVaxia ®) is limited by the serostatus of the vaccinee, and its use restricted to 9 to 45-year-old seropositive individuals. This project, developed in collaboration with the team of Dr. Ketloy of Chulalongkorn University (Thailand), aims at developing a safe and cost-effective tetravalent dengue vaccine protecting all individuals regardless of their age and serostatus at the time of vaccination. The developed vaccine should be easily produced, stable at fridge or room temperature and administered via intramuscular or mucosal route, with a simple dose-regimen. To achieve this goal, tetravalent adjuvanted nucleic acid lipid-based formulations are being developed and optimized using a Rational Design of Experiments approach.

Adjuvant research is of primary importance in the development of novel and safe vaccines against mutating infectious diseases. Pandemic influenza is a fitting example; in fact, researchers agree that the emergence of a new pandemic strain is not a question of if but a question of where and when. The most efficient way of controlling the rise and spread of a new pandemic disease is through the development of a safe and effective vaccine. Unfortunately, inactivated vaccines alone fail to induce a sufficient immune response and reuqire high doses and multiples administration, increasing vaccine campaign costs and reducing patient compliance. Adjuvants are necessary to ensure a sufficient immunogenicity. There is a need to identify new adjuvants or adjuvant combinations which can be combined with nano-carriers such as polymeric particles to increase targeting and thus reduce toxicity and allow dose sparing. In this study, we characterize and evaluate an adjuvant system composed of tomatine, solanine (lesser known saponins compared to Quillaja saponaria derivatives), SG101 NOD-like receptor (NLR) agonist, synthetized in the University of Ljubljana, encapsulated into PLGA-cholesterol nanoparticles for a WIV H5N1 influenza vaccine.

As a multifactorial disease, cancer imposes challenges that have been followed by constant effort by researchers to find new drugs, targets, and therapies. The cocktail combination used in this project was previously identified using the Therapeutically Guided Multidrug Optimization (TGMO) method. A four-drug combination called C2, composed of two tyrosine kinase inhibitors (TKIs): Tacedinaline and Tubacin with two histone deacetylase inhibitors (HDACIs): Erlotinib and Dasatinib was identified as a low-dose drug combination. The efficiency of an optimized drug cocktail is however hindered by distinct pharmacokinetic properties of each drug hence, solubility, polarity, charge, or stability. The result is a lack of control over the ratio, leading to the loss of synergy. Up to the challenge : the use of nanotechnology to load a drug cocktail  is a well-known strategy. Indeed, nanoparticles extend drug ’s blood circulation time and allow modulation of time and place of drug release. With their versatility and high biocompatibility, liposomal systems are the nanocarrier this project is based on.

Protein corona is an important factor in to predict nanomedicines efficacy and safety. Even though the corona has been characterized for different nanosystems, less is known about what happens to proteins at the nano-bio interface.

This project aims to predict and optimize the adsorption of a plasma protein to different surface functionalized superparamagnetic iron oxide nanoparticles (SPIONs). Transferrin was chosen as a protein model since it is generally detected on corona. Additionally, studies have shown that selective adsorption of the protein can alter the nanoparticles pharmacodynamics through specific interaction with its receptor. However, the mechanism of transferrin adsorption and potential conformation alterations has not yet been sufficiently examined.

We intend to describe and identify the key parameters involved in the nano-bio interface interactions, thus predicting the critical quality attributes (CQAs, i.e. interactions with plasma components) of the nanocarrier system. This will serve as a basis to engineer nanoparticles with a transferrin corona for specifically targeting transferrin receptor 1 (TfR1) expressing endothelia (e.g., the blood-brain barrier), or tumor cells with high metastatic potential.

Leading scientific and regulatory bodies in the pharmaceutical field acknowledged that the lack of established methods to provide reliable preclinical data represents the bottleneck in bringing promising nanomedicines to the market.  The aim of this project is to perform a thorough analysis of selected pharmaceutical nanosystems, focusing on the determination of size, shape, surface properties, and bio-nano interaction assessment (as selected combination of critical quality attributes - CQAs), applying a combination of orthogonal and complementary techniques. The overall impact of this project will lead to the reduction of risks, time, and financial investment in preclinical development of nanomedicines and “nanosimilars”, by suggesting new methods and optimized protocols for physicochemical and in vitro quality and safety assessment of these complex, innovative products. The project activities will gather information obtained from selected state-of-the-art methodologies, pointing out the minimal information level required for reliable nanomedicine characterization, bringing nanomedicines to a “more mature phase ”.

Retinal pathologies are diseases affecting the posterior segment of the eye, an area that is difficult to access for treatment. Intravitreal injection is the main route of administration because it directly targets the area of interest. However, this highly invasive administration has rare but serious side effects and is a heavy burden for the patient, who often stops the treatment early. It is therefore essential to look at other less invasive ocular delivery routes. This project focuses on the development of nanostructured lipid carriers, nanocapsules and nanoemulsions for topical application (eye drops). The selected raw materials, by promoting corneal absorption, increasing the residence time and solubilizing more API, allow to optimize ocular bioavailability and to reach the retina.

Immunotherapy sees its effectiveness limited by the resistance of poorly immunogenic “cold ”tumors, such as found in glioblastoma, prostate can or pancreatic cancer. In order to reduce this resistance, one strategy consists in modifying the tumor microenvironment (TME) towards the “hot ”phenotype, infiltrated by the immune system. Therefore, we used the activation of the stimulator of the interferon gene (STING) of the innate immunity pathway, targeting the cytosol of antigen-presenting cells (APC) in the TME. We opted for cGAMP, a negatively charged STING ligand. To allow targeting of APCs and efficient transfection to the cytosol of APCs (both dendritic cells (DC) and macrophages (M)) as well as protection against enzymes, positively charged carriers are required. For this, we formulated nanoparticles (NPs) with cGAMP through electrostatic interactions. To increase the quality of NP formulations, complementary techniques (DLS, NTA, AF4-MALS, UPLC-UV and EM) were used to characterize NPs ’critical quality attributes (CQA).

Intimal hyperplasia (IH) occurs in a considerable number of cases of blood vessel reconstruction by stenting or balloon angioplasty, venous bypass grafting, and arteriovenous dialysis accesses. It frequently leads to vessel occlusion and potential life-threatening graft failure.

To fight hyperplasia, we designed a delivery system for perivascular, local and sustained application of a statin. The device is capable of releasing the drug for several months thanks to microparticles reservoirs. The dopamine-derivatized hyaluronic acid gel that carries the microparticles may adhere to the tissue to ensure a long-lasting effect. Following successful proof-of-concept in in vitro and in mice, validation on larger models is foreseen towards an effective solution for the prevention of IH.

Osteoarthritis (OA) is the most common type of arthritis and degenerative joint disease. OA is a leading cause of chronic disability and progressively affects cartilage, the synovial membrane, bone and periarticular tissues . No efficient cure is yet available, and the current treatments essentially manage the symptoms, most often aiming at a reduction of inflammation and pain.

In an attempt to mitigate the progression of OA, the labs of pharmaceutical technology and biopharmacy are pursuing several approaches encompassing (i) the intra-articular administration  of sustained delivery microparticles releasing disease-modifying drugs, (ii) the design on novel hydrogels for the replacement/improvement of the worn-out synovial fluid and (iii) the administration of stem cells with concomitant nutrient supply.

The effectiveness of currently practised breast cancer treatment options usually does not last long enough and subsequently a new drug must be prescribed falling into a loop until there are no more treatment options available. Hence, a novel strategy is required. Our idea is to apply a microRNA cocktail, comprised of miRNAs that are downregulated in breast cancer, to restore their level of expression in the organism. Furthermore, in order to ensure efficient cell transfection, we are designing a polymer-based system to be implemented as a nanocarrier to overcome consecutive biological barriers and provide sustainable miRNA expression in the targeted region. In brief, we are working on a functional polymeric nanocarrier capable of encapsulating miRNAs which will constitute a potent treatment option to shrink breast cancer diagnosed at the early stage.

The burden of bacterial wound infections has considerably increased due to antibiotic resistance to most of the currently available antimicrobial drugs. Within this project, a new chemical platform for coupling chitosan derivatives to antimicrobial peptide dendrimers (AMPDs) has been developed. The AMPD-chitosan conjugates act synergistically in destroying the bacteria, without any antimicrobial activity loss. These new proprietary “active principles ”were successfully incorporated into various biopolymer formulations, including nanoparticles, gels, and foams. The antibacterial dressings are now tested in vivo in both healthy and infected mice. This chemical technological platform could be used for the development of new membrane disruptive therapeutics to eradicate pathogens present in acute and chronic wounds.