Treatment of spontaneous canine mast cell tumors by electrochemotherapy combined with IL-12 gene electrotransfer: Comparison of intratumoral and peritumoral application of IL-12

Tratar, Urša Lampreht; Milevoj, Nina; Čemažar, Maja; Žnidar, Katarina; Valentinuzzi, Katja Ursic; Brožič, Andreja; Tomsič, Katerina; Serša, Gregor; Tozon, Nataša

doi:10.1016/j.intimp.2023.110274

Cited by 8 publications

(15 citation statements)

References 49 publications

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“…The retention of plasmid DNA decreased significantly at 14 days, and at 28 days, plasmid DNA was present only in the plate (1 mg/mL and 2 mg/mL) groups. These results are similar to those gathered from veterinary studies, where the presence of plasmid DNA 7 days after the procedure was still evident but reduced over time and was almost nonexistent 28 days after the procedure [ 12 ]. Skin swab analysis from the phIL12 GET showed that the plasmid was not retained on the surface of the skin at the site of injection, irrespective of the electrodes used.…”

Section: Discussionsupporting

confidence: 87%

“…In addition, IL-12 reduces the growth of tumor vessels via IFNγ, which contributes to the antitumor effectiveness of IL-12 [ 8 ]. Studies have already been published demonstrating the antitumor efficacy of IL-12 in preclinical studies on mice [ 9 , 10 , 11 ], in clinical studies on spontaneous tumors in dogs [ 12 ], and in human studies on melanoma, where in addition to a local effect, they have also demonstrated a systemic effect on distant untreated tumors [ 13 ]. IL-12 GET has been predominantly used as an intratumoral or intradermal application [ 14 , 15 ].…”

Section: Introductionmentioning

confidence: 99%

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Safety and Efficacy of IL-12 Plasmid DNA Transfection into Pig Skin: Supportive Data for Human Clinical Trials on Gene Therapy and Vaccination

Lampreht Tratar,

Jesenko,

Omerzel

et al. 2024

IJMS

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Gene electrotransfer (GET) of plasmids encoding interleukin 12 (IL-12) has already been used for the treatment of various types of tumors in human oncology and as an adjuvant in DNA vaccines. In recent years, we have developed a plasmid encoding human IL-12 (phIL12) that is currently in a phase I clinical study. The aim was to confirm the results of a non-clinical study in mice on pharmacokinetic characteristics and safety in a porcine model that better resembled human skin. The GET of phIL12 in the skin was performed on nine pigs using different concentrations of plasmid phIL12 and invasive (needle) or noninvasive (plate) types of electrodes. The results of our study demonstrate that the GET of phIL-12 with needle electrodes induced the highest expression of IL-12 at the protein level on day 7 after the procedure. The plasmid was distributed to all tested organs; however, its amount decreased over time and was at a minimum 28 days after GET. Based on plasmid copy number and expression results, together with blood analysis, we showed that IL-12 GET is safe in a porcine animal model. Furthermore, we demonstrated that pigs are a valuable model for human gene therapy safety studies.

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Section: Discussionsupporting

confidence: 87%

Section: Introductionmentioning

confidence: 99%

Safety and Efficacy of IL-12 Plasmid DNA Transfection into Pig Skin: Supportive Data for Human Clinical Trials on Gene Therapy and Vaccination

Lampreht Tratar,

Jesenko,

Omerzel

et al. 2024

IJMS

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“…ECT and IL-12 GET were performed on canine mast cell tumors, a much better response to the treatment was obtained than with IL-12 p.t. GET resulted in prolonged disease- and progression-free intervals ( 37 ). In other studies ( 67 – 69 ), various spontaneous canine tumors were treated with a combination of ECT with IL-12 GET, with plasmid and chemotherapeutic drug injected concurrently and resulted in a good antitumor response in several different tumor types, except sarcoma.…”

Section: Discussionmentioning

confidence: 99%

“…Nevertheless, IFN-γ is also the main mediator of IL-12 systemic toxicity, as seen during clinical testing of recombinant IL-12 ( 28 ). Gene electrotransfer (GET) of plasmids encoding IL-12 has been shown to be an effective treatment approach, with controlled release and greater safety, as shown in preclinical settings in mouse models ( 29 – 32 ) and veterinary clinical studies in dogs ( 33 – 37 ). In addition, GET of plasmids encoding IL-12 is currently also in clinical trials, showing promising results ( 38 – 40 ).…”

Section: Introductionmentioning

confidence: 99%

The effectiveness of calcium electroporation combined with gene electrotransfer of a plasmid encoding IL-12 is tumor type-dependent

Lisec,

Markelc,

Ursic Valentinuzzi

et al. 2023

Front. Immunol.

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IntroductionIn calcium electroporation (CaEP), electroporation enables the cellular uptake of supraphysiological concentrations of Ca2+, causing the induction of cell death. The effectiveness of CaEP has already been evaluated in clinical trials; however, confirmatory preclinical studies are still needed to further elucidate its effectiveness and underlying mechanisms. Here, we tested and compared its efficiency on two different tumor models to electrochemotherapy (ECT) and in combination with gene electrotransfer (GET) of a plasmid encoding interleukin-12 (IL-12). We hypothesized that IL-12 potentiates the antitumor effect of local ablative therapies as CaEP and ECT.MethodsThe effect of CaEP was tested in vitro as well as in vivo in murine melanoma B16-F10 and murine mammary carcinoma 4T1 in comparison to ECT with bleomycin. Specifically, the treatment efficacy of CaEP with increasing calcium concentrations alone or in combination with IL-12 GET in different treatment protocols was investigated. We closely examined the tumor microenvironment by immunofluorescence staining of immune cells, as well as blood vessels and proliferating cells.ResultsIn vitro, CaEP and ECT with bleomycin reduced cell viability in a dose-dependent manner. We observed no differences in sensitivity between the two cell lines. A dose-dependent response was also observed in vivo; however, the efficacy was better in 4T1 tumors than in B16-F10 tumors. In 4T1 tumors, CaEP with 250 mM Ca resulted in more than 30 days of growth delay, which was comparable to ECT with bleomycin. In contrast, adjuvant peritumoral application of IL-12 GET after CaEP prolonged the survival of B16-F10, but not 4T1-bearing mice. Moreover, CaEP with peritumoral IL-12 GET modified tumor immune cell populations and tumor vasculature.ConclusionsMice bearing 4T1 tumors responded better to CaEP in vivo than mice bearing B16-F10 tumors, even though a similar response was observed in vitro. Namely, one of the most important factors might be involvement of the immune system. This was confirmed by the combination of CaEP or ECT with IL-12 GET, which further enhanced antitumor effectiveness. However, the potentiation of CaEP effectiveness was also highly dependent on tumor type; it was more pronounced in poorly immunogenic B16-F10 tumors compared to moderately immunogenic 4T1 tumors.

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“…electroporation, allows simultaneous delivery of drugs and genes to the tumors and produces a better antitumor response than when electrochemotherapy and peritumoral IL-12 gene electrotransfer were subsequently administered. Tumor control and survival of dogs after combined concomitant electrochemotherapy and IL-12 gene electrotransfer were improved and prolonged, respectively ( 40 ). In addition, a study on sarcoma bearing mice showed that adjuvant intramuscular mIL-12 application after initial electrochemotherapy is dose dependent and dependent on the amount of IL-12 in the system, achieving better results in immunocompetent mice ( 24 ).…”

Section: Intersection Of Electrochemotherapy and Immunotherapymentioning

confidence: 99%

Electrochemotherapy combined with immunotherapy – a promising potential in the treatment of cancer

Hadzialjevic,

Omerzel,

Trotovsek

et al. 2024

Front. Immunol.

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Electrochemotherapy is a novel, locoregional therapy that is used to treat cutaneous and deep-seated tumors. The electric pulses used in electrochemotherapy increase the permeability of the cell membranes of the target lesion and thus enhance the delivery of low-permeant cytotoxic drugs to the cells, leading to their death. It has also been postulated that electrochemotherapy acts as an in situ vaccination by inducing immunogenic cell death. This in turn leads to an enhanced systemic antitumor response, which could be further exploited by immunotherapy. However, only a few clinical studies have investigated the role of combined treatment in patients with melanoma, breast cancer, hepatocellular carcinoma, and cutaneous squamous cell carcinoma. In this review, we therefore aim to review the published preclinical evidence on combined treatment and to review clinical studies that have investigated the combined role of electrochemotherapy and immunotherapy.

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Treatment of spontaneous canine mast cell tumors by electrochemotherapy combined with IL-12 gene electrotransfer: Comparison of intratumoral and peritumoral application of IL-12

Cited by 8 publications

References 49 publications

Safety and Efficacy of IL-12 Plasmid DNA Transfection into Pig Skin: Supportive Data for Human Clinical Trials on Gene Therapy and Vaccination

Safety and Efficacy of IL-12 Plasmid DNA Transfection into Pig Skin: Supportive Data for Human Clinical Trials on Gene Therapy and Vaccination

The effectiveness of calcium electroporation combined with gene electrotransfer of a plasmid encoding IL-12 is tumor type-dependent

Electrochemotherapy combined with immunotherapy – a promising potential in the treatment of cancer

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