Evaluation and optimization of liver transfection with non-viral vectors in murine experimental Sepsis

• Grant number: 12/12270-0
• Support Opportunities: Scholarships in Brazil - Scientific Initiation
• Start date: October 01, 2012
• End date: September 30, 2014
• Field of knowledge: Health Sciences - Medicine - Medical Clinics
• Principal Investigator: Erich Vinicius de Paula
• Grantee: Rodolfo Monteiro Enz Hubert
• Host Institution: Faculdade de Ciências Médicas (FCM). Universidade Estadual de Campinas (UNICAMP). Campinas , SP, Brazil

Abstract

Sepsis is one of the greatest challenges in medical practice, with mortality rates as high as 70% in most serious cases. However, its treatment is based mainly on the use of antibiotics and supportive therapy. Unlike areas such as oncology and rheumatology, in which targeted-specific therapies are being increasingly utilized, in the sepsis context this type of strategy has not been incorporated into clinical practice yet. In the last years, a better understanding of sepsis pathophysiology allowed the discovery of potential therapeutic targets such as inflammatory cytokines, coagulation factors, and recently, endothelial barrier modulators. In the areas where targeted-specific therapy is largely disseminated, like oncology and inflammatory diseases, therapies are usually based on the use of recombinant proteins and monoclonal antibodies. Although effective, these strategies are expensive and require, in some cases, continuous infusion of proteins with a short half-life. In this context, gene therapy with non-viral vectors can be an attractive strategy, especially in sepsis. Indeed, sepsis has some characteristics that makes it attractive to gene therapy with non-viral vectors, such as the need for only transient expression, and the possibility that endothelial barrier breakdown present in sepsis might facilitate gene expression induced by these vectors. In this project, we intend to evaluate liver transfection with non-viral vectors in the context of murine experimental sepsis. Initial experiments will use the hydrodynamic transfection protocol, which has been shown to allow consistent liver expression with non-viral vectors but has not been studied in the context of sepsis. Next, we will study the impact of alterations of critical transfection parameters such as dose, dilution, and speed of injection, to evaluate whether sepsis results in any change in the pharmacological aspects of hydrodynamic transfection. All transfection efficiency experiments will use beta-galactosidase and? Green Fluorescent Protein? visualization in tissue sections. Knowledge of the behavior of hydrodynamic transfection in sepsis is fundamental for future preclinical studies using non-viral vectors in murine experimental sepsis, planned in our laboratory.(AU)