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Local delivery of anticancer agents or gene therapeutics to lung tumors can circumvent side effects or accumulation in non-target organs, but accessibility via the alveolar side of the blood-air barrier remains challenging. Polyplexes based on plasmid and linear polyethylenimine (LPEI) transfect healthy lung tissue when applied intravenously (i.v.) in the mouse, but direct delivery into the lungs results in low transfection of lung tissue. Nevertheless, LPEI could offer the potential to transfect lung tumors selectively, if accessible from the alveolar side. This study combined near infrared fluorescent protein 720 (iRFP720) and firefly luciferase as reporter genes for detection of tumor lesions and transfection efficiency of LPEI polyplexes, after intratracheal microspraying in mice bearing 4T1 triple negative breast cancer lung metastases. Simultaneous flow cytometric analysis of iRFP720 and enhanced green fluorescent protein expression in vitro demonstrated the potential to combine these reporter genes within transfection studies. Polyplex biophysics was characterized by single nanoparticle tracking analysis (NTA) to monitor physical integrity after microspraying in vitro. 4T1 cells were transduced with iRFP720-encoding lentivirus and evaluated by flow cytometry for stable iRFP720 expression. Growth of 4T1-iRFP720 cells was monitored in Balb/c mice by tomographic near infrared imaging, tissue and tumor morphology by computed tomography and magnetic resonance imaging. In 4T1-iRFP720 tumor-bearing mice, intratracheal administration of luciferase-encoding plasmid DNA by LPEI polyplexes resulted in successful tumor transfection, as revealed by bioluminescence imaging.

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A 28‐year‐old Icelandic horse gelding was presented with a laryngopharyngeal squamous cell carcinoma. The gelding had been treated for penile carcinoma in situ with a partial phallectomy 2 years earlier. Polymerase chain reaction of tumour DNA and subsequent amplicon sequencing revealed that the equine papillomavirus type 2 E6 oncogene sequences of both lesions were identical. There is strong evidence that equine papillomavirus type 2 is causally associated with genital squamous cell carcinomas and precancerous lesions. Recent reports indicate that equine papillomavirus type 2 might also play an active role in the pathogenesis of approximately 20% of equine squamous cell carcinomas in the oronasal, pharyngeal and laryngeal regions. To the authors’ knowledge, this is the first report of a horse consecutively developing a penile carcinoma in situ and a laryngopharyngeal squamous cell carcinoma that were apparently induced by the same equine papillomavirus type 2 variant. Possible equine papillomavirus type 2 infection pathways in this horse and the importance of early detection of lesions are discussed in this context.

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