Hyaluronidase and Collagenase Increase the Transfection Efficiency of Gene Electrotransfer in Various Murine Tumors

Abstract

One of the applications of electroporation/electropulsation in biomedicine is gene electrotransfer, the wider use of which is hindered by low transfection efficiency in vivo compared with viral vectors. The aim of our study was to determine whether modulation of the extracellular matrix in solid tumors, using collagenase and hyaluronidase, could increase the transfection efficiency of gene electrotransfer in histologically different solid subcutaneous tumors in mice. Tumors were treated with enzymes before electrotransfer of plasmid DNA encoding either green fluorescent protein or luciferase. Transfection efficiency was determined 3, 9, and 15 days posttransfection. We demonstrated that pretreatment of tumors with a combination of enzymes significantly increased the transfection efficiency of electrotransfer in tumors with a high extracellular matrix area (LPB fibrosarcoma). In tumors with a smaller extracellular matrix area and less organized collagen lattice, the increase was not so pronounced (SA-1 fibrosarcoma and EAT carcinoma), whereas in B16 melanoma, in which only traces of collagen are present, pretreatment of tumors with hyaluronidase alone was more efficient than pretreatment with both enzymes. In conclusion, our results suggest that modification of the extracellular matrix could improve distribution of plasmid DNA in solid subcutaneous tumors, demonstrated by an increase in transfection efficiency, and thus have important clinical implications for electrogene therapy.

Hyaluronidase Contributes to Early Inflammatory Events Induced by Electrotransfer in Mouse Skeletal Muscle

“…HYA enzyme is successfully used in applications other than vaccination. In mice it increases the efficiency of transfection by gene ET in solid subcutaneous tumors, and it has been shown to be effective for the spreading of chemotherapeutic agents in various cancer types (Spruss et al, 1995;Klocker et al, 1998;Cemazar et al, 2012).…”

An overview on therapeutic potential and various applications of microbial collagenases

“…There is a wide range of industrial applications of collagenase including food industries [74], cosmetic industries [75] tannery and meat industries, [76,77] but the most significant applications of collagenase is in the field of therapeutics. Collagenases have various noninvasive therapeutic applications such as treatment of Dupuytren's and Peyronie'sdisesase, burns, wound healing, intervertebral disc herniation, chronic total occlusions, glaucoma, cartilage repair, uterine fibroid, cellulite, keloid, nipple pain and degradation of human retained placenta [69,78] and cancer gene therapy [79,80]. The collagenases can mediate tumor invasion through several mechanisms, which include constitutive production of enzyme by the tumor cells, induction of collagenase production in the neighbouring stromal cells and interactions between tumor/ stromal cells to induce collagenase production by one or both cell types.…”

Tumor Stromal Disrupting Agent Enhances the Anticancer Efficacy of Docetaxel Loaded PEGylated Liposomes in Lung Cancer

“…As mentioned earlier, tumor extracellular matrix (ECM) is mainly composed of a dense collagen network embedded in the gel of glycosaminoglycans (GAGs), primarily hyaluronan. A number of studies demonstrating the use of collagenase, hyaluronidase and lysyl oxidase to degrade tumor ECM, as a means to improve the delivery of various therapeutics have been published in last decade [39,40]. Collagenase labeled gold nanoparticle exhibited 35% higher tumor accumulation compared with unlabeled gold nanoparticles [17].…”

“…Also, it has been shown that liposomes injected 1 h after the collagenase injection, led to approximately 1.5-fold increase in tumor uptake. Further, pretreatment with collagenase and hyaluronidase were found to enhance the transfection efficiency of plasmid DNA in subcutaneous tumor [39].…”