Use of the IL-1 receptor antagonist (Anakinra) on innate immunity: effect of brain death and preservation of the kidney in the cold

• Grant number: 20/13510-0
• Support Opportunities: Regular Research Grants
• Start date: August 01, 2021
• End date: January 31, 2024
• Field of knowledge: Health Sciences - Medicine - Medical Clinics
• Principal Investigator: Mario Abbud Filho
• Grantee: Mario Abbud Filho
• Host Institution: Faculdade de Medicina de São José do Rio Preto (FAMERP). Secretaria de Desenvolvimento Econômico (São Paulo - Estado). São José do Rio Preto , SP, Brazil

Abstract

Brain death (BD) triggers systemic inflammation, with consequent activation of inflammatory pathways in organs donated for transplants. Kidneys from deceased donors with extended criteria (ECD) or with high risk scores (KDPI> 85%) considered "non-ideal" kidneys, could be used, but have been discarded by Tx teams because they have inferior outcomes, worsening the shortage of organs for Tx. The prolonged duration of cold ischemia time (CIT) may also be involved in changing the inflammatory profile of these kidneys. In a previous study, we reported that molecules in the innate immunity pathways are more expressed in "non-ideal" kidneys, than in donor kidneys with standard criteria (SCD). One of these highly expressed molecules is interleukin IL-1² and based on this observation we elaborated the hypothesis that blocking this molecule could reduce the innate inflammatory response, easing sterile inflammation and improving the quality of "non-ideal" kidneys. Objectives: To evaluate the effect of administration of the IL-1 receptor antagonist (Anakinra) on gene and protein expression related to innate immunity and sterile inflammation, in rat kidneys, after the ME process and with prolonged cold ischemia time. Methods. Male Wistar rats will be used, divided into 4 groups: sham rats (n = 6), control rats with ME induction and without treatment (n = 6); 3) rats with induction of BD and administration of Anakinra in 2 different doses (0.5mg / kg (n = 6) and 1mg / kg (n = 6)) at the time of BD. After 6 hours of ventilation maintenance, the kidneys of the animals will be perfused and preserved with Euro-Collins solution and analyzed in the 12h and 24h cold preservation times. From the collected kidney samples, the extraction of total RNA for gene expression will be performed by real-time quantitative polymerase chain reaction (qPCR) using the TaqMan Gene Expression Array Plates system for molecules involved in the innate immune response in addition to histological analysis and immunohistochemistry. The descriptive statistical analysis will include absolute and relative frequencies for categorical variables and means, medians, standard deviation and variation for continuous variables. Comparison of groups of categorical variables will be performed using the chi-square test or Fisher's exact test, when appropriate. Values of P <0.05 will be considered significant. (AU)