title: Protein nanocages for self-triggered nuclear delivery of DNA-targeted chemotherapeutics in Cancer Cells.
abstract: A genetically engineered apoferritin variant consisting of 24 heavy-chain subunits (HFn) was produced to achieve a cumulative delivery of an antitumor drug, which exerts its cytotoxic action by targeting the DNA at the nucleus of human cancer cells with subcellular precision. The rationale of our approach is based on exploiting the natural arsenal of defense of cancer cells to stimulate them to recruit large amounts of HFn nanoparticles loaded with doxorubicin inside their nucleus in response to a DNA damage, which leads to a programmed cell death. After demonstrating the selectivity of HFn for representative cancer cells compared to healthy fibroblasts, doxorubicin-loaded HFn was used to treat the cancer cells. The results from confocal microscopy and DNA damage assays proved that loading of doxorubicin in HFn nanoparticles increased the nuclear delivery of the drug, thus enhancing doxorubicin efficacy. Doxorubicin-loaded HFn acts as a "Trojan Horse": HFn was internalized in cancer cells faster and more efficiently compared to free doxorubicin, then promptly translocated into the nucleus following the DNA damage caused by the partial release in the cytoplasm of encapsulated doxorubicin. This self-triggered translocation mechanism allowed the drug to be directly released in the nuclear compartment, where it exerted its toxic action. This approach was reliable and straightforward providing an antiproliferative effect with high reproducibility. The particular self-assembling nature of HFn nanocage makes it a versatile and tunable nanovector for a broad range of molecules suitable both for detection and treatment of cancer cells.
authors.affiliation: Dipartimento di Biotecnologie e Bioscienze, Università di Milano-Bicocca, Piazza della Scienza 2, 20126 Milano, Italy.
authors.forename: Michela
authors.initials: M
authors.lastname: Bellini
authors.affiliation: Dipartimento di Scienze Biomediche e Cliniche "Luigi Sacco", Università di Milano, Ospedale L. Sacco, Via G.B. Grassi 74, 20157 Milano, Italy. Electronic address: serena.mazzucchelli@gmail.com.
authors.forename: Serena
authors.initials: S
authors.lastname: Mazzucchelli
authors.affiliation: Dipartimento di Biotecnologie e Bioscienze, Università di Milano-Bicocca, Piazza della Scienza 2, 20126 Milano, Italy.
authors.forename: Elisabetta
authors.initials: E
authors.lastname: Galbiati
authors.affiliation: Dipartimento di Scienze Biomediche e Cliniche "Luigi Sacco", Università di Milano, Ospedale L. Sacco, Via G.B. Grassi 74, 20157 Milano, Italy.
authors.forename: Silvia
authors.initials: S
authors.lastname: Sommaruga
authors.affiliation: Dipartimento di Scienze Biomediche e Cliniche "Luigi Sacco", Università di Milano, Ospedale L. Sacco, Via G.B. Grassi 74, 20157 Milano, Italy.
authors.forename: Luisa
authors.initials: L
authors.lastname: Fiandra
authors.affiliation: Dipartimento di Scienze Biomediche e Cliniche "Luigi Sacco", Università di Milano, Ospedale L. Sacco, Via G.B. Grassi 74, 20157 Milano, Italy.
authors.forename: Marta
authors.initials: M
authors.lastname: Truffi
authors.affiliation: Dipartimento di Biotecnologie e Bioscienze, Università di Milano-Bicocca, Piazza della Scienza 2, 20126 Milano, Italy.
authors.forename: Maria A
authors.initials: MA
authors.lastname: Rizzuto
authors.affiliation: Dipartimento di Biotecnologie e Bioscienze, Università di Milano-Bicocca, Piazza della Scienza 2, 20126 Milano, Italy.
authors.forename: Miriam
authors.initials: M
authors.lastname: Colombo
authors.affiliation: Dipartimento di Biotecnologie e Bioscienze, Università di Milano-Bicocca, Piazza della Scienza 2, 20126 Milano, Italy.
authors.forename: Paolo
authors.initials: P
authors.lastname: Tortora
authors.affiliation: Dipartimento di Scienze Biomediche e Cliniche "Luigi Sacco", Università di Milano, Ospedale L. Sacco, Via G.B. Grassi 74, 20157 Milano, Italy.
authors.forename: Fabio
authors.initials: F
authors.lastname: Corsi
authors.affiliation: Dipartimento di Biotecnologie e Bioscienze, Università di Milano-Bicocca, Piazza della Scienza 2, 20126 Milano, Italy. Electronic address: davide.prosperi@unimib.it.
authors.forename: Davide
authors.initials: D
authors.lastname: Prosperi
mesh_terms: D000970:Antineoplastic Agents / Q000008:administration & dosage* / Q000737:chemistry; D001052:Apoferritins / Q000008:administration & dosage* / Q000737:chemistry; D045744:Cell Line, Tumor; D002467:Cell Nucleus / Q000187:drug effects*; D004249:DNA Damage; D018076:DNA, Complementary / Q000008:administration & dosage / Q000494:pharmacology; D004273:DNA, Neoplasm / Q000187:drug effects*; D004926:Escherichia coli / Q000378:metabolism; D006367:HeLa Cells; D006801:Humans; D058990:Molecular Targeted Therapy / Q000379:methods*; D049329:Nanostructures / Q000737:chemistry*; D014178:Translocation, Genetic
chemical_list: D000970:Antineoplastic Agents; D018076:DNA, Complementary; D004273:DNA, Neoplasm; D001052:Apoferritins
keywords: Anticancer drugs; Apoferritin; Bionanoparticles; Drug delivery system; Nuclear targeting; Self-triggered translocation