The impetus for new curative approaches for HIV-AIDS lacks adequate clinical tools to monitor patient responses to therapeutics targeting the persistent viral reservoir. We therefore propose to develop a standardizable, integrated, clinically-relevant whole blood culture assay to evaluate HIV eradication therapeutics ex vivo by directly measuring responses of primary peripheral blood cells under internally- referenced patient-specific matched control and therapeutic conditions. The proposed system will quantify single-cycle virus production from infected primary blood cells comprising the entire peripheral blood immunological compartment. The relative difference in virus production under latency-activating conditions versus unstimulated matched cultures represents the target replication-competent provirus residing in latently- infected primary blood cells. The culture assay will support ex vivo testing of latency-activating, cytolytic, immunomodulatory and other curative therapeutic approaches required by the FDA for therapeutic success. This standardizable, readily-accessible whole blood culture assay for testing HIV eradication strategies will provide an enabling, next-generation research, clinic and patient-management platform. The Jericho team proposes to validate the feasibility of developing such a platform here via this ambitious Phase I SBIR project (expanded scope and budget). The envisioned integrated blood-collection system for HIV-infected whole blood culture is a closed device requiring less than 10.0 mL of donor blood per collection tube, directly obtained by a phlebotomist using a standard blood draw. After up to 72 hours of incubation at 37?C in a proprietary media, viable cells, soluble factors and virions can be co-collected and measured from these primary blood cultures to provide correlative biomarkers. The proposed product will allow for comparison of different test conditions (e.g., latency activators) and therapeutics (e.g., suppressive drugs, cytokines) across multiple internally-referenced culture tubes, specific to the infection status and immunological response of the individual. We propose to demonstrate the feasibility of standardizing and developing this platform to evaluate endpoints for HIV eradication strategies by characterizing virus production, cellular markers, and soluble factors from 45 HIV-infected donor whole blood sample collections. We will first quantify and characterize reproducibility of our stimulated virus production culture system within a coefficient of variance ?14% between replicate, matched collection tubes. We will then demonstrate the potential utility of the characterized platform using two selected compounds known to stimulate latently-infected cells. Preclinical therapeutic optimization translates directly to clinical monitoring and patient management, having identified relevant conditions and associated biomarkers using the proposed whole blood culture assay platform. As HIV is now moving from a manageable chronic disease to a cure, novel diagnostics will be increasingly required to ensure that the best treatments are used to optimize efficacy and to reduce patient risks.
This Phase I SBIR project is designed to prove the feasibility of developing a comprehensive, whole blood culture platform useful for clinically testing therapeutics intended for eradication of HIV-1 infection. This work represents a key step toward producing a next-generation research and clinical platform that will enable the optimization of therapeutic treatments for individuals infected with HIV-1, together with its complications. In thi way, the proposed technology meets NIH's mission to treat immune-mediated diseases, to develop new and more-effective treatments for HIV-infected people, and to find a cure. Specifically, the technology supports NIAID HIV Cure Research to: (1) determine the effects of latency reversal, immune modulation, cell therapies, and other interventions on the HIV reservoir; (2) study HIV latency in the context of clinically-infected whole blood cultures; and (3 investigate strategies that lead to purging of the latent reservoir and a cure for HIV/AIDS or control of viral rebound and sustained remission.
