SERTOLI CELLS AS CARRIERS FOR GENE THERAPY VECTORS IN BRAIN AND CANCER

ANTONELA S ASAD; PATRICIA JACOBO; CRISTIAN SOBARZO; SOLEDAD MENDEZ; FLAVIA ZANETTI; MARIELA A. MORENO AYALA; CAMILA ZUCATTO; MARÍA VERÓNICA BAEZ; DIANA JERUSALINSKY; ADRIANA SEILICOVICH; SUSANA THEAS; LIVIA LUSTIG; MARIANELA CANDOLFI

Mar del Plata

Congreso; LXI Reunión Científica Anual de la Sociedad Argentina de Investigación Clínica (SAIC), LXIV Reunión de la Sociedad Argentina de Inmunología (SAI), XLVIII Reunión Anual de la Sociedad Argentina de Farmacología Experimental (SAFE).; 2016

Since Sertoli cells are key regulators in testis immunoprivilege, they constitute good candidates for cell-mediated gene therapy. Viral vectors are excellent tools for the delivery of therapeutic transgenes in chronic diseases. However, these vectors could be immunogenic leading to transient expression in vivo. Our hypothesis was that Sertoli cells could be useful carriers for the delivery of immunogenic gene therapy vectors into the brain parenchyma and the tumor microenvironment. Sertoli cells were obtained by enzymatic digestion (90% purity) from prepuberal or adult mice testis and infected with adenoviral (Adv) vectors (MOI 15) encoding the green fluorescent protein (GFP) under the CMV promoter. Transduction efficiency was assessed in vitro by fluorescent microscopy. Sertoli cells were harvested for in vivo injection 24 h after infection. Naïve mice received intracranial injections of Adv-infected Sertoli cells (60,000). Infected syngeneic Sertoli cells (35,000) were also injected in breast tumors growing in the flank of Balb/c mice or into glioblastomas growing in the brain of C57Bl/6 mice. In Sertoli cells from prepuberal mice, transgene expression was detected in 38% of cells 24 h post-infection. Transduction efficiency was higher when cells were obtained from adult mice (90%) in comparison with prepuberal mice (60%). GFP+ Sertoli cells from prepuberal or adult mice were readily detected in the normal brain parenchyma 72 h after injection. However, brains injected with adult-derived Sertoli cells exhibited higher frequency of GFP+ cells. Adult-derived GFP+ Sertoli cells were also detected within the tumor microenvironment in breast and brain tumors. CONCLUSIONS: Our results show that Sertoli cells from adult animals exhibit more stable transgene expression than prepuberal animals, probably due to their low proliferation rate. Sertoli cells could be useful carriers for the delivery of therapeutic transgenes for chronic diseases.