Stem cell-based delivery of TRAIL (tumor necrosis factor-related apoptosis-inducing ligand) therapy aiming at blocking renal fibrosis in a model of experimental chronic kidney disease

• Grant number: 21/00091-2
• Support Opportunities: Scholarships in Brazil - Post-Doctoral
• Effective date (Start): June 01, 2021
• Effective date (End): November 09, 2023
• Field of knowledge: Biological Sciences - Physiology - Physiology of Organs and Systems
• Principal Investigator: Irene de Lourdes Noronha
• Grantee: Felipe Mateus dos Santos Ornellas
• Host Institution: Faculdade de Medicina (FM). Universidade de São Paulo (USP). São Paulo , SP, Brazil

Abstract

Tubulointerstitial fibrosis is the final common pathway leading to end stage Chronic Kidney Disease (CKD), ultimately with renal replacement therapy. So far, there is no approved anti-fibrotic therapy for this situation. Myofibroblasts, effector cells of renal fibrosis, contribute to the establishment and progression of fibrogenesis. Myofibroblasts have a particularly high expression of Death Receptors (DR), which make them sensitive to Tumor necrosis factor-related Apoptosis-Inducing Ligand (TRAIL) inducing cell death. TRAIL is a transmembrane protein that induces apoptosis via death receptors, playing an important role in the regulation of cell survival and inflammation. Selective apoptosis induced by exogenous TRAIL through DRs signaling expressed by tumor cells, sparing the normal ones, emerged the use of TRAIL as a strategy for an anticancer molecular target. This selectivity is mainly based on the expression of DR4 and DR5. The successful of TRAIL as anticancer therapy has triggered interest to investigate the role of TRAIL in other cells that undergo phenotypic and morphological differentiation, such as fibroblasts, which undergo transdifferentiation into myofibroblasts, opening up a line of investigation in fibrogenesis. In this context, strategies to block or even eradicate myofibroblasts represent an interesting alternative to prevent and/or reverse renal fibrogenesis. Diabetic mice knockout for TRAIL presented increased proteinuria, fibrosis and renal inflammation. In diabetic nephropathy experimental models, the treatment with recombinant TRAIL improved the morphological changes and blocked the expression of pro-fibrotic genes. The antifibrotic effects appear to be related to the high DR expression in myofibroblasts, making them sensitive to cell death induced by TRAIL. In fact, preliminary results developed during my sandwich doctorate at the Center for Nanomedicine - Johns Hopkins University, showed an increase in DR5 mRNA in renal myofibroblasts. The short half-life of recombinant TRAIL, however, limits the success of exogenous TRAIL. To overcome these limitations, the present project aims to induce the expression of TRAIL through gene therapy. For this purpose, stem cells will be used as a TRAIL gene vector to enable delivery and to allow the induction of TRAIL protein synthesis in renal tissue. This strategy becomes innovative and promising for the blocking/reversal of fibrosis due to potential anti-fibrotic action of TRAIL associated with the renoprotective effects induced by stem cells already described. The Folic Acid Nephropathy (FAN) model, recently standardized in our laboratory, will be used. Thus, the objective of the present study is to analyze the effect of the combination of cellular and gene therapy, via transfer of TRAIL gene carried by adipose cells derived Mesenchymal Stem Cells (MSC-AD), in a model of fibrogenic kidney disease. Specific objectives include: 1) to evaluate the efficiency of TRAIL gene transfer in vitro in renal fibroblasts, and in vivo, in the FAN model; 2) verify the efficacy of therapy with TRAIL gene delivered by MSC-AD as prevention of kidney disease, performed in zero time, at the time of induction of the FAN model in order to prevent/attenuate the development of fibrosis; 3) to analyze the possible effects of reversal of renal fibrosis through the administration of therapy with TRAIL gene delivered by MSC-AD, after the establishment of renal fibrosis, better simulating the conditions observed in the clinical practice of CKD. The study aims to demonstrate that stem cell-based delivery of TRAIL therapy can be established as a new therapy for renal fibrosis through of selective myofibroblast apoptosis, as well the simultaneous negative regulation of multiple fibrotic markers important for installation and maintenance of renal fibrosis. (AU)