The proposed project integrates groups from academia and industry to develop and perform initial testing of a novel platform for the potential eradication of HIV-1 by endowing the patient's own cells with stable resistance to HIV infection. To safely and effectively achieve such resistant cells, we propose to deploy a process call genome editing to either (i) perform targeted insertion of anti-HIV gene sequences, and/or permanently disrupt HIV relevant gene targets, directly in the genome of the patient's hematopoietic stem/progenitor cells (HSPC). Genome editing is achieved by invoking the cell's own natural DNA repair pathways to heal a double-strand break (DSB) introduced in vivo at the target locus by an engineered DNA cleavage enzyme called a zinc finger nuclease (ZFN). This technology has recently entered the clinic in subjects with HIV, where the safety, tolerability and early evidence of genetic protection from HIV infection has been observed following an infusion of autologous CCR5 gene edited CD4+ T cells. While CCR5 is a validated target for development of HIV therapeutics, disruption of this gene in isolation will not confer resistance to dual or X4-tropic HIV isolates. To expand the potential application of genome edited HSPCs to achieve a potential cure for HIV our strategy is perform the genetic addition of anti-HIV gene products to a defined location in the stem cell genome. Specifically, we propose to develop targeted nucleases to enable the in situ editing of endogenous human restriction factors as well develop methods to achieve the site-specific addition of anti-HIV genes in the critical long term hematopoietic stem cell (LT-HSC). The latter has recently been identified as a key area for optimization of genome editing strategies in these promising therapeutic stem cells.

Public Health Relevance

The proposed project integrates efforts from groups in industry and academia to develop and perform initial testing of new genome edited stem cell products aimed at achieving a cure for HIV. This work aims to overcome the basic science and translational obstacles to efficient site-specific addition of anti-HIV genes in to the critical long term hematopoietic stem cell (LT-HSC).

National Institute of Health (NIH)
National Heart, Lung, and Blood Institute (NHLBI)
Research Program--Cooperative Agreements (U19)
Project #
Application #
Study Section
Special Emphasis Panel (ZAI1-BP-A (J3))
Project Start
Project End
Budget Start
Budget End
Support Year
Fiscal Year
Total Cost
Indirect Cost
University of Southern California
Los Angeles
United States
Zip Code
Canny, Marella D; Moatti, Nathalie; Wan, Leo C K et al. (2018) Inhibition of 53BP1 favors homology-dependent DNA repair and increases CRISPR-Cas9 genome-editing efficiency. Nat Biotechnol 36:95-102
Colonna, Lucrezia; Peterson, Christopher W; Schell, John B et al. (2018) Evidence for persistence of the SHIV reservoir early after MHC haploidentical hematopoietic stem cell transplantation. Nat Commun 9:4438
Paul, Biswajit; Ibarra, Guillermo S Romano; Hubbard, Nicholas et al. (2018) Efficient Enrichment of Gene-Modified Primary T Cells via CCR5-Targeted Integration of Mutant Dihydrofolate Reductase. Mol Ther Methods Clin Dev 9:347-357
Reeves, Daniel B; Peterson, Christopher W; Kiem, Hans-Peter et al. (2017) Autologous Stem Cell Transplantation Disrupts Adaptive Immune Responses during Rebound Simian/Human Immunodeficiency Virus Viremia. J Virol 91:
Peterson, Christopher W; Benne, Clarisse; Polacino, Patricia et al. (2017) Loss of immune homeostasis dictates SHIV rebound after stem-cell transplantation. JCI Insight 2:e91230
Wang, Cathy X; Cannon, Paula M (2016) The clinical applications of genome editing in HIV. Blood 127:2546-52
Peterson, Christopher W; Wang, Jianbin; Norman, Krystin K et al. (2016) Long-term multilineage engraftment of autologous genome-edited hematopoietic stem cells in nonhuman primates. Blood 127:2416-26
Wang, Jianbin; Exline, Colin M; DeClercq, Joshua J et al. (2015) Homology-driven genome editing in hematopoietic stem and progenitor cells using ZFN mRNA and AAV6 donors. Nat Biotechnol 33:1256-1263