Bioengineered of T- and NK-cells by CAR against invasive fungal infections

Abstract

Invasive fungal infections (IFIs) show mortality at similar and higher levels compared to tuberculosis and malaria. Regarding neglected diseases, IFIs have a low financial support while tuberculosis and malaria are better supported. Since IFIs, candidiasis, aspergilosis, criptococosis and histoplasmosis have more effect on the global public health. The risk factors of IFIs are related to immunocompromised host over many situations, but previous data showed growth rate of immunocompetent host affected by IFIs. In this case, the virulence factors of Cryptococcus spp, C. albicans and H. capsulatum are responsible to promote the escape from host immune response. The CD4+ T cells can differentiate in T helper (Th) 1 (Th1) and 17 (Th17), Tc1 and Tc17 profiles originated from CD8+ T cells and the cytotoxic effects of natural killer (NK) cells, which can be impair to reach the infection site. Then, the current project aims to redirect T and NK cells by chimeric antigen receptor (CAR) promoting the direct effect of these immune cells in the infection site affected by IFIs. This new strategy of cell therapy against IFIs was started by Laurence J.N. Cooper's and Kumaresan's lab, which added into CAR the carbohydrate recognition domain of Dectin-1 (DectinCAR). Basically, CAR has four regions: (I) binding domain, (II) hinge or stalk region, (III) transmembrane region, (IV) cytoplasmic signaling domain. Cooper's and Kumar's lab created T cells modified expressing DectinCAR that showed a therapeutic effect against Aspergillus fumigatus. This strategy therapeutic was realized for the responsible the current project that trained in these leader's groups of CAR, between 2017 and 2018 supported by FAPESP/BEPE, and I developed a CAR T cells to target Cryptococcus spp. These factors together support the aim of this current project, and three steps are essentials: (I) Phage Display technique to generate scFv (single chain variable fragment) specific to target C. gattii, C. neoformans, C. albicans or H. capsulatum; (II) construction of lentiviral vector to express a CAR specific for each fungi, and to do the transduction of the Jurkat cells to evaluate the recognition and activation triggered by CAR in the presence of the target; (III) redirection of CAR T cells and CAR NK cells for each fungi, and to analyze the effector activity of these modified cells against in vitro and in vivo infection by C. gattii, C. neoformans, C. albicans ou H. capsulatum. The current proposal has strong evidence that redirected T and NK cells by CAR to target fungi into the infection site can mediate a protective immune response against IFIs. Thus, this project opens perspectives for clinical application with high impact in the scientific field, and some steps from this project can produce a patent process due the relevance of CAR T cells in the cell therapy. Keywords: T cells; NK cells; Invasive fungal infections; CAR; Cell therapy. (AU)