NANOTECHNOLOGICAL APPROACHES FOR CANCER DIAGNOSIS, TREATMENT AND MANAGEMENT: IMPLEMENTATION OF FERRITIN PROTEIN NANOPARTICLES FOR TUMOR TARGETING

Despite the efforts performed in the field of cancer research, data concerning the incidence of this pathology in North America and Europe reveal that tumors still represent the second cause of death. In fact, the effectiveness of many drugs is often seriously affected by lack of specificity in hitting the therapeutic target, determining the onset of toxicity issues. In this context, Nanomedicine aims to make a decisive contribution. Nanoparticle (NPs)-based drug delivery systems can enhance the physicochemical properties of a wide variety of drugs used in oncology to limit off-site side effects and improve their therapeutic efficacy, increasing drug accumulation in target tissue. Thus, through the development of nanocarriers capable of selectively recognizing and interacting with tumor cells and/or tissues, potential groundbreaking applications in the diagnosis, treatment and prevention of cancer could become feasible. In my PhD project, I exploited the unique features of a NPs based on recombinant heavy-chain ferritin nanocages (HFn). HFn nanocages represent a promising protein- based class of NPs for drug delivery widely investigated by virtue of (1) their natural tumor homing and limited accumulation in off-target organs which leads to reduced off-target toxicity; (2) their biocompatibility; (3) their structure that allows the encapsulation of different types of drugs or fluorescent tracers; (4) their versatility to be modified at the surface both exploiting chemical and genetical strategies. My work has been divided into three main studies aimed at implementing innovative strategies based on HFn nanocages both for diagnostic and therapeutic approaches in oncology . In Study 1, I presented the exploitation of HFn nanocages as nanodevices for fluorescence image-guided surgery (FGS). This technique is gaining interest due to its potential to improve tumor margin visualization and real-time identification of tumor deposits, maximizing the benefits for patients. However, no tumor-targeted probe effectively capable of localizing tumor tissue is available to date. For this reason, we proposed a HFn-based nanotracer for the delivery of indocyanine green (ICG), which is the main fluorescent dye currently used in clinics for FGS. A novel variant of HFn was developed, extensively characterized in vitro and loaded with ICG for in vivo experiments demonstrating potentiality as tumor-targeted nanotracers for the identification of tumors. 3 In Study 2, HFn nanocages were exploited for the encapsulation of a chemotherapeutic drug, doxorubicin (DOX) obtaining FerOX nanoformulation. FerOX has been investigated by virtue of its natural tumor homing and limited accumulation in off- target organs which leads to reduced off-target toxicity of drugs herein incapsulated. In particular, the object of this study was to assess the efficacy of FerOX as an alternative formulation capable of preserving the proliferative potential of human lymphocytes and the potential impact in the generation of an adaptative immune response after treatment. In Study 3, we focused on the use of HFn nanocages conjugated to the monoclonal anti-HER2 antibody trastuzumab (TZ). By exploiting the ability of HFn to cross the blood brain barrier (BBB), we achieved the delivery of TZ as a prophylactic treatment to tackle brain metastasis in HER2-positive advanced breast cancer.