Antimicrobial peptides loaded inhalable calcium phosphates nanoparticles for the counteraction of antibiotic resistance: towards a new therapy for respiratory infections

Principal investigator: Michele Iafisco

Personnel involved: Camilla Fusacchia, Federica Mancini

Starting date: 25/04/2022
Duration: 36 months
Total funding: 430.340,00 €
CNR-ISSMC role: Partner
Coordinator: Maurizio Sanguinetti (Università Cattolica del Sacro Cuore)
Consortium: Università Cattolica del Sacro Cuore; Consiglio Nazionale delle Ricerche (CNR-ISSMC, CNR-SCITEC e CNR-IRGB)

Antibiotic resistance (AMR) is one of the top 10 global public health threats facing humanity. In the last two decades, respiratory bacteria have increasingly become resistant to several antibiotics, and the prevalence of resistant strains is growing rapidly. Hospital-acquired pneumonia, ventilator-acquired pneumonia, and chronic respiratory diseases such as cystic fibrosis are some examples of respiratory diseases complicated by infections with multidrug resistant pathogens. The search for new antibiotic formulations for these infections is therefore a primary objective in the current health context. Antimicrobial peptides (AMPs) are emerging category of therapeutic agents with a huge potential application. These molecules have a broad spectrum of action and differently from the usually employed drugs, act by different mechanisms against which bacteria hardly develop resistance mechanisms. However, one of the main concerns is related to stability of AMPs after administration, which dramatically may reduce their power. Additionally, the efficacy of providing a sustained localization at specific site might not be properly achieved. In this view, the use of nanoparticles (NPs) for the AMPs delivery provides different advantages: (i) release of molecule in a more specific time and spatial window, and (ii) peptide protection against early degradation. Therefore, a multifunctional NP-based therapeutic formulation certainly represents an effective alternative to traditional therapies. The aim of AppliCare is to reach the proof of concept for a new therapeutic formulation based on inhalable and biodegradable calcium phosphate (CaPs) NPs, functionalized with new AMPs. CaP-NPs were selected for this project because are inhalable and fully biocompatible. As first step, an in-silico identification, production, and cytotoxicity assessment of novel peptides with putative antimicrobial activity and inhibition of ribosome assembly will be carried out. Then, the evaluation of the in vitro antimicrobial and antibiofilm activity and the behavior of the selected AMPs in clinical pathogens will be performed. The best-in-class AMPs will be used for the functionalization of CaP-NPs and the in vitro antimicrobial and antibiofilm activity of the formulations will be tested. The biodistribution and biocompatibility of the lead formulation will be assessed in healthy animals, as well as its efficacy in one selected animal model of respiratory infection. The success of our approach will pave the way for an effective bypassing of the multidrug resistance burden in pulmonary infections. AppliCare is multidisciplinary and 2 research units (Università Cattolica del Sacro Cuore and Consiglio Nazionale delle Ricerche) with different and complementary scientific backgrounds including molecular and cellular biology, microbiology, biochemistry, chemistry, material sciences, biophysics, in silico modeling, and animal physiology, are involved for the achievement of the overall goal.

Publications and patents

• Antimicrobial peptides for tackling cystic fibrosis related bacterial infections: A review. Francesca Bugli, Cecilia Martini, Maura Di Vito, Margherita Cacaci, Daniele Catalucci, Alessandro Gori, Michele Iafisco, Maurizio Sanguinetti, Alberto Vitali. Microbiological Research, 263, 2022, 127152.

• Biocompatible antimicrobial colistin loaded calcium phosphate nanoparticles for the counteraction of biofilm formation in cystic fibrosis related infections. Iafisco, M.; Carella, F.; Degli Esposti, L.; Adamiano, A.; Catalucci, D.; Modica, J.; Bragonzi, A.; Vitali, A.; Torelli, R.; Sanguinetti, M.; Bugli, F  Journal of Inorganic Biochemistry 2022, 230, 111751.