Biodistribution and Safety Assessment of Bladder Cancer Specific Recombinant Oncolytic Adenovirus in Subcutaneous Xenografts Tumor Model in Nude Mice

Abstract

Background The previous works about safety evaluation for constructed bladder tissue specific adenovirus are poorly documented. Thus, we investigated the biodistribution and body toxicity of bladder specific oncolytic adenovirus Ad-PSCAE-UPII-E1A (APU-E1A) and Ad-PSCAE-UPII-E1A-AR (APU-E1A-AR), providing meaningful information prior to embarking on human clinical trials. Materials and Method Conditionally replicate recombinant adenovirus (CRADs) APU-E1A, APU-EIA-AR were constructed with bladder tissue specific Uroplakin II (UP II) promoter to induce the expression of Ad5E1A gene and E1A-AR fusing gene, and PSCAE was inserted at upstream of promoter to enhance the function of promoter. Based on the cytopathic and anti-tumor effect of bladder cancer, these CRADs were intratumorally injected into subcutaneous xenografts tumor in nude mice. We then determined the toxicity through general health and behavioral assessment, hepatic and hematological toxicity evaluation, macroscopic and microscopic postmortem analyses. The spread of the transgene E1A of adenovirus was detected with RT-PCR and Western blot. Virus replication and distribution were examined with APU-LUC administration and Luciferase Assay. Results General assessment and body weight of the animals did not reveal any alteration in general behavior. The hematological alterations of groups which were injected with 5×108 pfu or higher dose (5×109 pfu) of APU-E1A and APU-E1A-AR showed no difference in comparison with PBS group, and only slight increased transaminases in contrast to PBS group at 5×109 pfu of APU-E1A and APU-E1A-AR were observed. E1A transgene did not disseminate to organs outside of xenograft tumor. Virus replication was not detected in other organs beside tumor according to Luciferase Assay. Conclusions Our study showed that recombinant adenovirus APU-E1A-AR and APU-E1A appear safe with 5×107 pfu and 5×108 pfu intratumorally injection in mice, without any discernable effects on general health and behavior.

Replicon RNA Viral Vectors as Vaccines

“…An extensive comparison to other delivery systems is not within the scope of this review, so only a few examples are addressed. Clearly, adenovirus-based vaccine development and gene therapy has a longer history, which has generated a multitude [130] of vector improvements and also resulted a number of clinical trials [131,132]. Similarly, herpes simplex virus (HSV) vectors have been frequently applied and HSV-GM-CSF have, for instance, been subjected to phase I−III human clinical trials in glioblastoma and melanoma patients [133].…”

Expression and functional evaluation of Mytilus galloprovincialis foot protein type 5 (Mgfp-5), the recombinant mussel adhesive protein

“…The common method is to implant the test material into the body of an animal. However, implantation in vivo is limited by the long experimental period, complicated operation process, complex body environment and parameter control, among others (Wang et al, 2012;Lee et al, 2016). In contrast, experiments conducted in vitro are relatively simple and their reproducibility can easily be controlled (Li et al, 2015).…”

YB-1-basierte Virotherapie

“…immunogenen Zelltods der Tumorzelle [6]. Vacciniaviren und Reoviren entwickelt und charakterisiert [7,8,9,10,11,12,13,14]. Im Design dieser Viren wurden dabei unterschiedliche Strategien hinsichtlich der Replikationskontrolle und des therapeutischen Effekts entwickelt.…”