Combined Ferritin Nanocarriers with ICG for Effective Phototherapy Against Breast Cancer

Leopoldo Sitia,1,* Paola Saccomandi,2,* Leonardo Bianchi,2 Marta Sevieri,1 Cristina Sottani,3 Raffaele Allevi,1 Elena Grignani,3 Serena Mazzucchelli,1 Fabio Corsi1,4 1Department of Biomedical and Clinical Sciences, Università degli studi di Milano, Milan, Italy; 2Department of Mechanical Engineering, Politecnico di Milano, Milan, Italy; 3Environmental Research Center, Istituti Clinici Scientifici Maugeri IRCCS, Pavia, Italy; 4Istituti Clinici Scientifici Maugeri IRCCS, Pavia, Italy*These authors contributed equally to this workCorrespondence: Serena Mazzucchelli, Department of Biomedical and Clinical Sciences, Università degli studi di Milano, via G.B. Grassi 74, Milan, 20157, Italy, Email serena.mazzucchelli@unimi.it Fabio Corsi, Istituti Clinici Scientifici Maugeri IRCCS, via S. Maugeri 10, Pavia, 27100, Italy, Email fabio.corsi@unimi.itIntroduction: Photodynamic Therapy (PDT) is a promising, minimally invasive treatment for cancer with high immunostimulatory potential, no reported drug resistance, and reduced side effects. Indocyanine Green (ICG) has been used as a photosensitizer (PS) for PDT, although its poor stability and low tumor-target specificity strongly limit its efficacy. To overcome these limitations, ICG can be formulated as a tumor-targeting nanoparticle (NP).Methods: We nanoformulated ICG into recombinant heavy-ferritin nanocages (HFn-ICG). HFn has a specific interaction with transferrin receptor 1 (TfR1), which is overexpressed in most tumors, thus increasing HFn tumor tropism. First, we tested the properties of HFn-ICG as a PS upon irradiation with a continuous-wave diode laser. Then, we evaluated PDT efficacy in two breast cancer (BC) cell lines with different TfR1 expression levels. Finally, we measured the levels of intracellular endogenous heavy ferritin (H-Fn) after PDT treatment. In fact, it is known that cells undergoing ROS-induced autophagy, as in PDT, tend to increase their ferritin levels as a defence mechanis