Combination HAART therapy, which predominantly targets the activities ofthe HIV-1 reverse transcriptase, protease, antj integrase enzymes, works in the majority of cases to inhibit HIV-1 replication and slow or stop the development of AIDS in infected patients. Despite the resounding success of HAART, drug resistance arises in a significant number of treated patients. There is therefore a continual need to develop so-called multi-generational drugs that work in the face of viral drug resistance mutations, as well as new compounds that inhibit HIV-1 through entirely novel means. Recent years have highlighted the interactions between host cellular proteins and viral proteins as novel drug targets, and this grant application previously established the critical role that the interaction between the cellular LEDGF/p75 protein and integrase plays in directing HIV- 1 to integrate into active genes. Ongoing work will ascertain the mechanisms of LEDGF/p75 binding to cellular chromatin and details ofthe interaction between LEDGF/p75 and the viral pre-integration complex. Work conducted during the current funding cycle moreover clarified that small molecule allosteric integrase inhibitors that compete with LEDGF/p75 for binding to integrase rather remarkably inhibited HIV-1 replication at the point of viral core maturation, an unprecedented finding for inhibitors of integrase catalytic function. Such compounds will now be used to further clarify the role of LEDGF/p75 in the virus replication cycle. The work will also determine the roles of novel integrase binding proteins in HIV-1 replication and the division of labor among integrase protomers in the pre-integration complex, which in turn could lead to new avenues of antiviral inhibitor development. The grant recently identified gain-of-function mutants in the HIV-1 integrase that enhanced significantly the transduction of target cells that express reverse charged, complementary LEDGF/p75 proteins, which opened the avenue to targeted gene therapy vectors in human cells. Ongoing work will continue to optimize retroviral DNA integration targeting through the modification of host cell proteins. The successful completion of these studies will refine the mechanisms of action of known targets of antiretroviral drugs, discover new targets for inhibitor development, and invent ways to introduce transgenes.

Public Health Relevance

The work will clarify the mechanism of action of known antiretroviral drugs and uncover new targets for small molecule inhibitor development. Technologies to direct the integration of retroviral vectors will improve the safety of genetic transduction.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Method to Extend Research in Time (MERIT) Award (R37)
Project #
4R37AI039394-20
Application #
8755871
Study Section
Special Emphasis Panel (NSS)
Program Officer
Miller, Roger H
Project Start
2015-06-01
Project End
2020-05-31
Budget Start
2015-06-01
Budget End
2016-05-31
Support Year
20
Fiscal Year
2015
Total Cost
Indirect Cost
Name
Dana-Farber Cancer Institute
Department
Type
DUNS #
076580745
City
Boston
State
MA
Country
United States
Zip Code
Lummertz da Rocha, Edroaldo; Rowe, R Grant; Lundin, Vanessa et al. (2018) Reconstruction of complex single-cell trajectories using CellRouter. Nat Commun 9:892
Engelman, Alan N; Singh, Parmit K (2018) Cellular and molecular mechanisms of HIV-1 integration targeting. Cell Mol Life Sci 75:2491-2507
El Ashkar, Sara; Schwaller, Juerg; Pieters, Tim et al. (2018) LEDGF/p75 is dispensable for hematopoiesis but essential for MLL-rearranged leukemogenesis. Blood 131:95-107
Hoyte, Ashley C; Jamin, Augusta V; Koneru, Pratibha C et al. (2017) Resistance to pyridine-based inhibitor KF116 reveals an unexpected role of integrase in HIV-1 Gag-Pol polyprotein proteolytic processing. J Biol Chem 292:19814-19825
Sugimura, Ryohichi; Jha, Deepak Kumar; Han, Areum et al. (2017) Haematopoietic stem and progenitor cells from human pluripotent stem cells. Nature 545:432-438
Alvarez, Frances J D; He, Shaoda; Perilla, Juan R et al. (2017) CryoEM structure of MxB reveals a novel oligomerization interface critical for HIV restriction. Sci Adv 3:e1701264
Lesbats, Paul; Engelman, Alan N; Cherepanov, Peter (2016) Retroviral DNA Integration. Chem Rev 116:12730-12757
Lu, Yi-Fen; Cahan, Patrick; Ross, Samantha et al. (2016) Engineered Murine HSCs Reconstitute Multi-lineage Hematopoiesis and Adaptive Immunity. Cell Rep 17:3178-3192
Serrao, Erik; Cherepanov, Peter; Engelman, Alan N (2016) Amplification, Next-generation Sequencing, and Genomic DNA Mapping of Retroviral Integration Sites. J Vis Exp :
Serrao, Erik; Engelman, Alan N (2016) Sites of retroviral DNA integration: From basic research to clinical applications. Crit Rev Biochem Mol Biol 51:26-42

Showing the most recent 10 out of 39 publications