A Novel Method for Controlled Gene Expression via Combined Bleomycin and Plasmid DNA Electrotransfer

Abstract

Electrochemotherapy is an efficient method for the local treatment of cutaneous and subcutaneous metastases, but its efficacy as a systemic treatment remains low. The application of gene electrotransfer (GET) to transfer DNA coding for immune system modulating molecules could allow for a systemic effect, but its applications are limited because of possible side effects, e.g., immune system overactivation and autoimmune response. In this paper, we present the simultaneous electrotransfer of bleomycin and plasmid DNA as a method to increase the systemic effect of bleomycin-based electrochemotherapy. With appropriately selected concentrations of bleomycin and plasmid DNA, it is possible to achieve efficient cell transfection while killing cells via the cytotoxic effect of bleomycin at later time points. We also show the dynamics of both cell electrotransfection and cell death after the simultaneous electrotransfer of bleomycin and plasmid DNA. Therefore, this method could have applications in achieving the transient, cell death-controlled expression of immune system activating genes while retaining efficient bleomycin mediated cell killing.

Tetrahedral Framework Nucleic Acids Reverse New-Onset Type 1 Diabetes

“…The induction of regulatory immune cells to treat or prevent T1D is an exceptionally promising immunotherapy for T1D . Instead of infusing regulatory immune cells directly, inducing them and halting the destructions of β-cells through making use of biomaterials are more favored. − In many studies, systemic administration of biomaterials can accurately induce regulatory immune cells, delete autoreactive T-cells, re-establish immune tolerance, and meanwhile maintain the function and structure of the islets. , Among the biomaterials, nanoscale materials account for the vast majority. ,− As one of the promising nanoscale materials, various forms of DNA nanomaterials are usually adopted in immune therapies, such as DNA nanogels, DNA nanocapsules, and DNA origami. − In immunotherapy, the DNA nanomaterials themselves were unable to modulate the immune system in most cases. , The DNA nanomaterials are commonly treated as vehicles to deliver therapeutics to the body areas that other delivery systems cannot reach …”

The Evidence of the Bystander Effect after Bleomycin Electrotransfer and Irreversible Electroporation

“…It is also known that one bleomycin molecule can make multiple DNA lesions [ 42 ]. In our recent publication, we demonstrated that DNA degradation takes place after bleomycin electrotransfer at concentrations used in the present paper [ 43 ]. Since ECT, like radiation therapy, is a local treatment associated with local cell damage, ROS generation, DNA damage (in case of bleomycin electrotransfer), and release of DAMPs, the bystander effect following ECT seems to be very plausible ( Figure 5 ).…”

Effects of buffer composition and plasmid toxicity on electroporation-based non-viral gene delivery in mammalian cells using bursts of nanosecond and microsecond pulses

“…4), also less popular buffers from other studies focused on nucleofection are included to enable better repeatability and consolidation of knowledge (e.g., No. 3, 6–8) ( Pakhomov et al, 2014 ; Parreno et al, 2015 ; Zhou et al, 2016 ; Chopra et al, 2019 ; Gibot et al, 2020 )…”