Therapeutic Efficacy of the Novel Stimuli-Sensitive Nano-Ferritins Containing Doxorubicin in a Head and Neck Cancer Model

Abstract

Doxorubicin is employed alone or in combination for the treatment of several hematological and solid malignancies; despite its efficacy, there are associated cardiotoxicity limits both in its application in patients with heart disease risk factors and also in its long-term use. HFt-MP-PAS40 is a genetically engineered human ferritin heavy chain (HFt)-based construct able to efficiently entrap and deliver doxorubicin to cancer cells. HF-MP-PAS contains a short motif sequence (defined as MP) responsive to proteolytic cleavage by tumor matrix metalloproteases (MMPs), located between each HFt subunit and a masking polypeptide sequence rich in proline (P), alanine (A), and serine (S) residues (PAS). This carrier displayed excellent therapeutic efficacy in a xenogenic pancreatic cancer model in vivo, leading to a significant increase in overall animal survival in treated mice. Herein, we describe the HFt-MP-PAS40-Dox efficacy against squamous cell carcinomas of the head and neck (HNSCC) with the goal of validating the application of our nano-drug for the treatment of different solid tumors. In addition, a tolerability study in healthy mice was also performed. The results indicate that HFt-MP-PAS40-Dox produced increased anti-tumor effects both in vitro and in vivo in comparison to the free drug in several HNSCC cell lines. In the acute toxicity studies, the maximum tolerated dose (MTD) of HFt-MP-PAS40-Dox was about 3.5 higher than the free drug: 25 mg/kg versus 7 mg/kg doxorubicin equivalents. Importantly, evaluation of heart tissues provided evidence that doxorubicin is less cardio-toxic when encapsulated inside the ferritin carrier. In conclusion, HFt-MP-PAS40-Dox may be administered safely at higher doses compared with the free drug, resulting in superior efficacy to control HNSCC malignancies.

Engineered Human Nanoferritin Bearing the Drug Genz-644282 for Cancer Therapy

“…PAS and PASE polypeptides enhance HFt water-solubility and determine a 10/15-fold drop in CD71 binding affinity, as determined by surface plasmon resonance experiments [ 24 ]. Decreased binding to the bulk of CD71 molecules expressed by a multitude of normal tissues, albeit at low levels, both extends the half-life of the nanocarrier in the bloodstream as compared to native HFt [ 23 , 24 , 25 , 26 ] and increases the nanocarrier chances to recirculate until it binds at highly dense in CD71, but dimensionally small tumor site. Once there, the PASE-linked MP sequence is selectively and conditionally cleaved off by matrix metalloproteases 2 and 9, which are greatly enriched in the tumor microenvironment but essentially absent in normal tissues, leading to HFt unmasking and local restoration of HFt:CD71 binding and tumor killing.…”

“…Once there, the PASE-linked MP sequence is selectively and conditionally cleaved off by matrix metalloproteases 2 and 9, which are greatly enriched in the tumor microenvironment but essentially absent in normal tissues, leading to HFt unmasking and local restoration of HFt:CD71 binding and tumor killing. The HFt-MP-PASE construct proved indeed to be capable of delivering the canonical anticancer drugs doxorubicin and mitoxantrone (Mit), both of which were stably entrapped in the internal HFt cavity, to human pancreatic (PaCa44) and head and neck (FaDu) tumor xenotransplants in vivo, with good therapeutic efficacy [ 23 , 24 , 26 ]. However, complete and durable tumor regression could not be obtained either by us using these payloads, or by other groups using different HFt:drug nanosystems [ 2 , 23 , 26 ].…”

Doxorubicin loaded ferritin nanoparticles for ferroptosis enhanced targeted killing of cancer cells

“…[18][19][20] Nanomaterials have shown to great potential in encapsulating and transporting drugs, penetrating cell membranes and releasing drugs in tumor cells through the enhanced permeability and retention (EPR) effect, 21 which augment the accumulation of drugs in tumor cells but reduce that in normal cells. 22 Ferritin (Fn), a universal intracellular protein that stores iron and releases it in a controlled fashion, was designed to encapsulate DOX for targeting delivery drugs to transferrin receptor 1 (TFR1) overexpressed tumor cells, 23,24 human leukemia (HL-60), human colorectal carcinoma (Lovo), human breast adenocarcinoma (MDA-MB-468), human liver carcinoma (Hep G2), human cervical adenocarcinoma (HeLa), human breast (MCF-7), human small cell lung carcinoma (NCI-H69). 25,26 Ferritin, with DOX loaded, could directly interact with TFR1 (ref.…”

“…By this way, DOX was released in tumor cells for therapeutic research. 24 In this study, DOX was loaded into iron saturated ferritin. The ferritin was designed as a tumor targeting drug carrier.…”

Ferritin Nanocages for Protein Delivery to Tumor Cells

“…By "protein cage", we mean a structure of protein nature consisting of subunits that self-assemble to form a highly symmetric hollow nanosphere large enough to enclose therapeutics such as conventional drugs, nucleic acids, and other proteins. In addition to viral capsids, excellent examples of protein cages are thermosomes [32], small heat-shock proteins [33], pyruvate dehydrogenase multienzyme complexes [34], and ferritins [35][36][37][38][39][40]. Although all these proteins have been extensively studied for the delivery of conventional drugs and/or contrast agents, very little has been done regarding their use in the delivery of therapeutic proteins.…”