Ex Vivo Profiling and Targeting of Metabolic Dysregulation in HIV-1 Infected Lymph Node Organoids.

The major obstacle to a cure for HIV-1 infection is the presence of viral reservoirs formed by latently infected CD4 T cells, in which the

virus is transcriptionally silent and thus hidden from the host immune response and shielded from current antiretroviral therapies. To

overcome this barrier, it is essential to find a cellular signature of latent HIV-1 infection allowing the detection and the selective

targeting of latently infected cells. However, the identification of specific markers of latency has proven so far difficult and few in vitro

studies have translated into clinical trials. One potential reason is the difficulty to recapitulate with standard 2D in vitro cultures the

complex tissue environment in which the viral reservoir is mainly seeded in vivo, the lymph node. In addition, the direct study of

lymph nodes is hampered by the need of an invasive procedure for their isolation. This project is aimed at leveraging my expertise in

working with 3D immune cell cultures to study latency markers of infection in a tonsil-derived organoid system that closely

recapitulates the lymph node environment while conserving the tunability and practicality of standard in vitro systems. With this tool,

I will focus the research of latency markers on cellular metabolic imbalances induced by HIV-1 infection. This line of study is pursued

by my host lab and has already been proven promising, having led to the reduction of the viral reservoir in a macaque model and in a

phase IIa clinical trial. I plan to obtain a comprehensive picture of the metabolic landscape of latently infected CD4 T cells in a

lymphoid organoid, which will guide the identification of selective markers of latency. The organoid system will then be employed to

test therapeutic candidates, which will be identified by in silico screenings, for their ability to kill latently infected cells in their native environments: the lymph node organoids and finally in lymphoid tissues of people living with HIV.