Proof of Concept of a Local PROteoLIposomes-based Therapy Against Acute Lungs Infection in Chronic Granulomatous Disease Patients – PROLIC

Chronic Granulomatous Disease(CGD) is a rare genetic disease of innate immunodeficiency affecting phagocytes (neutrophils, macrophages) of patients by dysfunction of the membrane NADPH oxidase complex. This enzyme is responsible for the production of microbicidal toxic oxygen derivatives (ROS). The most frequent genetic form is the X-linked CGD (XCGD) affecting the NOX2/p22phox redox element of the NADPH oxidase complex. Thus, XCGD patients suffer from severe and recurrent infections, the primary cause of death being acute lung infection refractory to intravenous anti-infectious treatments. Only an infusion of granulocytes to boost their defective immunity can save them but this strongly compromises a possible subsequent allogenic marrow transplant and this treatment is also difficult to organize in an emergency context. Thus, a local treatment applied in emergency has all its place. The main objective of this project is to make a proof of concept of the efficacy and the safety of a local protein-based therapy using proteoliposomes in experimental models of acute lung infections, in a CGD context.
Recombinant membrane NOX2/p22phox complex will be expressed in HEK293 TRex cells after lentiviral infection. Affinity purification will be done by adding a "tag" to the plasmid construct. The PLs will be constituted by a lipidic mixture containing phosphocholine azido allowing to link by "click" chemistry molecules for the specific targeting of PLs to pulmonary macrophages. The proof of concept of the efficacy of native or modified PLs will be done by analyzing the restoration of the NADPH oxidase activity in NOX2/p22phox deficient cell models such as XCGD macrophages derived from induced pluripotent cells of patients. Finally, the efficacy of the optimized PLs will be tested in a XCGD mouse model of acute lung infections.