T-cell based assay to identify novel drug targets against HIV-1-envelope processing

Includes bibliographical references (p. 25-27).It is well established that HIV relies on both the viral protease and host proteases as part of its life cycle. The only poly-protein processed by host enzymes is the viral envelope, which is processed by Furin and similar peptidases and protein convertases (mainly PC-1) that reside in the luminal face of the Endoplasmic Reticulum (ER). The main goal of this defense is the development of a specific cell-based assay aimed at targeting the host cellular-based processing of the HIV envelope. The assay will allow the monitoring of the HIV-1 envelope processing, which is based on the cleavage of the gp160 envelope protein precursor, resulting in gp120 and gp41 products. Our assay is based on the engineering of a scaffold protein, which will travel to the cell surface unless HIV-1 envelope processing is blocked or inhibited. This will be detected by flow cytometry, allowing to discriminate between successful and inhibited or blocked envelope processing. Importantly, the assay, once implemented, will allow screening of ER/Golgi-localized random peptide libraries for the search of a novel family of antivirals. These antivirals will target the recognition of gp160 by Furin or similar peptidases rather that inhibit their activity, which would probably be detrimental to the cell. Moreover, the assay will be developed in T-cells, a cell type readily infected by HIV, through retroviral technology. Importantly, this assay promises to shed light on specific processe