Development of WRAP5 Peptide Complexes for Targeted Drug/Gene Co-Delivery toward Glioblastoma Therapy

Abstract

Despite the great progress over the past few decades in both the diagnosis and treatment of a great variety of human cancers, glioblastoma remains the most lethal brain tumor. In recent years, cancer gene therapy focused on non-viral vectors which emerged as a promising approach to glioblastoma treatment. Transferrin (Tf) easily penetrates brain cells of the blood–brain barrier, and its receptor is highly expressed in this barrier and glioblastoma cells. Therefore, the development of delivery systems containing Tf appears as a reliable strategy to improve their brain cells targeting ability and cellular uptake. In this work, a cell-penetrating peptide (WRAP5), bearing a Tf-targeting sequence, has been exploited to condense tumor suppressor p53-encoding plasmid DNA (pDNA) for the development of nanocomplexes. To increase the functionality of developed nanocomplexes, the drug Temozolomide (TMZ) was also incorporated into the formulations. The physicochemical properties of peptide/pDNA complexes were revealed to be dependent on the nitrogen to phosphate groups ratio and can be optimized to promote efficient cellular internalization. A confocal microscopy study showed the capacity of developed complexes for efficient glioblastoma cell transfection and consequent pDNA delivery into the nucleus, where efficient gene expression took place, followed by p53 protein production. Of promise, these peptide/pDNA complexes induced a significant decrease in the viability of glioblastoma cells. The set of data reported significantly support further in vitro research to evaluate the therapeutic potential of developed complexes against glioblastoma.

Evidence That a Peptide-Drug/p53 Gene Complex Promotes Cognate Gene Expression and Inhibits the Viability of Glioblastoma Cells

“…The preparation of TMZ-loaded peptides was already described in a previous publication from our team [ 43 ]. After synthesis and TMZ loading, peptides were suspended in ultrapure water and kept at −20 °C.…”

“…The morphology of the peptide/pDNA complexes developed at N/P ratios of 0.5 and 1 was determined by scanning electron microscopy (SEM). The microscopy images have been reported in a previous publication [ 43 ].…”

“…In this context, the design/development of non-viral delivery systems to promote cellular uptake and intracellular targeted payload delivery has been considered a convenient tool to increase therapeutic efficacy in cancer gene therapy [ 38 , 39 , 40 ]. Cell-penetrating peptides (CPPs) are short peptides composed of less than 30 amino acids that can carry biomolecules and easily translocate through the cell membrane—a property related to the amino acid sequence [ 41 , 42 , 43 ]. Although the mechanism of CPP penetration is not fully understood, it seems to be influenced by factors such as the cell type, membrane composition, properties of the cargo, or peptide/cargo ratio [ 44 , 45 , 46 ].…”

« New Modalities in Cancer Imaging and Therapy » XVe édition de l’atelier organisé par le réseau « Vectorisation, Imagerie, Radiothérapies » du Cancéropôle Grand-Ouest, 5–8 octobre 2022, Erquy, France

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Targeted Glioblastoma Treatment via Synthesis and Functionalization of Gold Nanoparticles With De Novo–Engineered Transferrin-Like Peptides: Protocol for a Novel Method

“…Given the potential synergy among AuNPs, transferrin-like peptides, temozolomide, and MGMT inhibitors, we propose the synthesis, stabilization, and functionalization of AuNPs with de novo–engineered transferrin-like peptides and the aforementioned chemotherapeutic agents [ 7 , 13 , 14 ]. This exploratory proposal aims to outline a novel protocol for a potentially more effective GBM treatment strategy, offering insights and considerations for future researchers in this field.…”