Peptide-Targeted Polyplexes for Aerosol-Mediated Gene Delivery to CD49f-Overexpressing Tumor Lesions in Lung

• Published in: Molecular therapy. Nucleic acids Vol. 18; pp. 774 - 786
• Main Authors: Taschauer, Alexander, Polzer, Wolfram, Alioglu, Fatih, Billerhart, Magdalena, Decker, Simon, Kittelmann, Theresa, Geppl, Emanuela, Elmenofi, Salma, Zehl, Martin, Urban, Ernst, Sami, Haider, Ogris, Manfred
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 06.12.2019; Elsevier Limited; American Society of Gene & Cell Therapy; Elsevier
• Subjects: Alveoli; Antigens; Bioluminescence; Breast cancer; CD49f targeting; Cell surface; Chromatography; Cytology; Gene expression; Gene transfer; Internalization; Lesions; Ligands; linear polyethylenimine; Localization; Lung cancer; lung tumor transfection; Lungs; Magnetic resonance imaging; Metastasis; Molecular weight; Nanoparticles; Particle size; Peptides; Polyethylene glycol; polyplexes; Proteins; pulmonary delivery; Reporter gene; Software; Statistical analysis; Stem cells; Surface markers; Trachea; Tumor cell lines; Tumor cells; Tumors; United States > US; Austria; Germany; linear polyethylenimine; pulmonary delivery; lung tumor transfection; CD49f targeting; polyplexes
• ISSN: 2162-2531; 2162-2531
• DOI: 10.1016/j.omtn.2019.10.009

Peptide ligands can enhance delivery of nucleic acid-loaded nanoparticles to tumors by promoting their cell binding and internalization. Lung tumor lesions accessible from the alveolar side can be transfected, in principle, using gene vectors delivered as an aerosol. The cell surface marker CD49f (Integrin α6) is frequently upregulated in metastasizing, highly aggressive tumors. In this study, we utilize a CD49f binding peptide coupled to linear polyethylenimine (LPEI) promoting gene delivery into CD49f-overexpressing tumor cells in vitro and into lung lesions in vivo. We have synthesized a molecular conjugate based on LPEI covalently attached to the CD49f binding peptide CYESIKVAVS via a polyethylene glycol (PEG) spacer. Particles formed with plasmid DNA were small (<200 nm) and could be aerosolized without causing major aggregation or particle loss. In vitro, CD49f targeting significantly improved plasmid uptake and reporter gene expression on both human and murine tumor cell lines. For evaluation in vivo, localization and morphology of 4T1 murine triple-negative breast cancer tumor lesions in the lung of syngeneic BALB/c mice were identified by MRI. Polyplexes applied via intratracheal aerosolization were well tolerated and resulted in measurable transgene activity of the reporter gene firefly luciferase in tumor areas by bioluminescence imaging (BLI). Transfectability of tumors correlated with their accessibility for the aerosol. With CD49f-targeted polyplexes, luciferase activity was considerably increased and was restricted to the tumor area.