HIV persists for many years during therapy and normally rebounds to pretreatment levels when therapy is stopped. However, a small number of individuals have been able to control their infection, despite the presence of infectious virus, fo many years after stopping therapy. Understanding the factors that lead to virus control under these circumstances is critical to our ability to achieve this end in others as well as for an eventual cure of infection. We are learning that where the virus integrates its genome into the infected cell genome is of significance to the maintenance of HIV reservoirs. However, during suppressive therapy, the rarity of cells with replication competent HIV and the lack of expression of viral proteins by these cells have hampered the study of HIV reservoirs. We propose novel methods to overcome the problem of cell rarity by the use of a novel method to enrich for and expand single infected cells for study of structure and RNA expression of the integrated proviral genome and surrounding sequences within the host cell genome. To address the problem of the lack of protein expression, we will use additional novel methods to purify expanded infected cells, as lysates, allowing analysis of the entire cellular transcriptome and proteome. Lastly, we will recreate the proviral structures we observe in vivo in an in vitro model to discern the global impact of provirus integration on cell functionality.

Public Health Relevance

We propose to understand the role HIV has, as it exists at different sites in the infected cell genome, to the persistence of HIV reservoirs in different cell types and tissues, and the ability to control HIV infection after interruption of ART. We propose 3 Aims to address these issues, each building on the previous, and each providing heretofore unprecedented windows into reservoir cell structure, function and virologic control.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
5R01AI125026-03
Application #
9528449
Study Section
Special Emphasis Panel (ZRG1)
Program Officer
Kuo, Lillian S
Project Start
2016-08-11
Project End
2020-07-31
Budget Start
2018-08-01
Budget End
2019-07-31
Support Year
3
Fiscal Year
2018
Total Cost
Indirect Cost
Name
University of Washington
Department
Microbiology/Immun/Virology
Type
Schools of Medicine
DUNS #
605799469
City
Seattle
State
WA
Country
United States
Zip Code
98195