Duration of the project:
Partners:
Institute of Oncology Ljubljana
University of Primorska Faculty of Health Sciences
University of Ljubljana Veterinary Faculty
Principal investigator / researcher:
Maja Čemažar, PhD (SICRIS, ResearchGate)
Project coordinator at UP FHS:
Maja Čemažar, PhD (SICRIS, ResearchGate)
Link:
Summary:
Scientific background and problem statement: In the last decades immunogene therapy for the treatment of cancer is gaining in importance. Currently there are 59 running immunegene studies in the field of human oncology, 11 of them are using plasmid DNA encoding the cytokine interleukin 12 (IL-12) indicating the great potential of this kind of therapy. In spite of marked advances in the field of gene therapy, the problem of safety and efficiency of this therapy in combination with standard treatment modalities remains. In the process of translation of these therapies to human medicine, the importance of clinical studies in veterinary medicine is becoming more and more valued, especially in dogs, namely spontaneously occurring tumors in dogs and the similarity of canine and human immune systems represent an important translational bridge accelerating the transition to human medicine.
The aim of the proposed project is the development and validation of a safer adjuvant immune gene therapy with IL-12 in combination with surgery for the treatment of oral and skin tumors in dogs. For this purpose plasmid DNA encoding canine IL-12 gene under transcriptional control of the tissue specific promoter for skin will be constructed. Plasmid will be evaluated in cell cultures and in tumor models in vivo. Clinical protocol will be prepared, and all necessary approvals will be obtained from the competent authorities and first patients will be recruited into the clinical study.
Methods: Plasmids encoding canine IL-12 under the transcriptional control of a tissue specific promoter and with kanamycin resistance gene will be prepared using standard molecular biology methods. Plasmid without the antibiotic resistance gene will be prepared according to the instructions of the supplier of ORT® technology. Appropriate bacterial strains will be transformed with the plasmids to produce larger quantities of endotoxin free plasmid to be used in further experiments. The expression profiles of the plasmids will be determined in different canine and human cells using RT-PCR and ELISA. Additionally proliferation assay will be performed. Antitumor effect of the prepared plasmids will be determined in canine tumor xenografts in nude mice. Antitumor effectiveness will be assessed by tumor growth delay assay and tumor control probability assay and histological analysis of the tumors will be conducted. Clinical study will be executed at the Clinic for Surgery and Small Animal of the Veterinary faculty University of Ljubljana.
Execution and management of the project: The work on the project will be executed at the laboratories of 5 partners participating in the project, where all necessary equipment for molecular biology, cell cultures, immunocytochemistry, animal experiments and clinical study is available. The researchers involved in the project have vast experience in the field of plasmid preparation, cell culture isolation, electroporation as a drug and gene delivery method, clinical protocols preparation, and electroporation in veterinary patients.
The management and coordination of the project will be assured by regular meetings and constant communication with partners on the project.
Relevance and potential impact of the results: The proposed project is designed as a preclinical and clinical project that addresses important issues regarding safe use of cancer gene therapy in clinical setting. The results of the project, if successful, may have impact on implementation of gene therapy in veterinary oncology and above all the translation of such clinical studies into human oncology in Slovenia and worldwide.
»The authors acknowledge the project J3-6796 Translational oncology: development and validation of immunogene therapy with Interleukin-12 combined with surgery for canine cancers of the oral cavity and skin was finacially supported by the Slovenian Research Agency.«
