title: Engineering HIV-Resistant, Anti-HIV Chimeric Antigen Receptor T Cells.
abstract: The treatment or cure of HIV infection by cell and gene therapy has been a goal for decades. Recent advances in both gene editing and chimeric antigen receptor (CAR) technology have created new therapeutic possibilities for a variety of diseases. Broadly neutralizing monoclonal antibodies (bNAbs) with specificity for the HIV envelope glycoprotein provide a promising means of targeting HIV-infected cells. Here we show that primary human T cells engineered to express anti-HIV CARs based on bNAbs (HIVCAR) show specific activation and killing of HIV-infected versus uninfected cells in the absence of HIV replication. We also show that homology-directed recombination of the HIVCAR gene expression cassette into the CCR5 locus enhances suppression of replicating virus compared with HIVCAR expression alone. This work demonstrates that HIV immunotherapy utilizing potent bNAb-based single-chain variable fragments fused to second-generation CAR signaling domains, delivered directly into the CCR5 locus of T cells by homology-directed gene editing, is feasible and effective. This strategy has the potential to target HIV-infected cells in HIV-infected individuals, which might help in the effort to cure HIV.
authors.affiliation: Center for Immunity and Immunotherapies and Program for Cell and Gene Therapy, Seattle Children's Research Institute, Seattle, WA 98101, USA.
authors.forename: Malika
authors.initials: M
authors.lastname: Hale
authors.affiliation: Center for Global Infectious Disease Research, Seattle Children's Research Institute, Seattle, WA 98101, USA.
authors.forename: Taylor
authors.initials: T
authors.lastname: Mesojednik
authors.affiliation: Center for Immunity and Immunotherapies and Program for Cell and Gene Therapy, Seattle Children's Research Institute, Seattle, WA 98101, USA.
authors.forename: Guillermo S
authors.initials: GS
authors.lastname: Romano Ibarra
authors.affiliation: Center for Immunity and Immunotherapies and Program for Cell and Gene Therapy, Seattle Children's Research Institute, Seattle, WA 98101, USA.
authors.forename: Jaya
authors.initials: J
authors.lastname: Sahni
authors.affiliation: Center for Global Infectious Disease Research, Seattle Children's Research Institute, Seattle, WA 98101, USA.
authors.forename: Alison
authors.initials: A
authors.lastname: Bernard
authors.affiliation: Center for Immunity and Immunotherapies and Program for Cell and Gene Therapy, Seattle Children's Research Institute, Seattle, WA 98101, USA.
authors.forename: Karen
authors.initials: K
authors.lastname: Sommer
authors.affiliation: Center for Immunity and Immunotherapies and Program for Cell and Gene Therapy, Seattle Children's Research Institute, Seattle, WA 98101, USA; Department of Pediatrics, University of Washington, Seattle, WA 98101, USA; Department of Immunology, University of Washington, Seattle, WA 98101, USA.
authors.forename: Andrew M
authors.initials: AM
authors.lastname: Scharenberg
authors.affiliation: Center for Immunity and Immunotherapies and Program for Cell and Gene Therapy, Seattle Children's Research Institute, Seattle, WA 98101, USA; Department of Pediatrics, University of Washington, Seattle, WA 98101, USA; Department of Immunology, University of Washington, Seattle, WA 98101, USA. Electronic address: drawling@uw.edu.
authors.forename: David J
authors.initials: DJ
authors.lastname: Rawlings
authors.affiliation: Center for Global Infectious Disease Research, Seattle Children's Research Institute, Seattle, WA 98101, USA; Department of Pediatrics, University of Washington, Seattle, WA 98101, USA. Electronic address: thor.wagner@seattlechildrens.org.
authors.forename: Thor A
authors.initials: TA
authors.lastname: Wagner
mesh_terms: D057134:Antibodies, Neutralizing / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D003602:Cytotoxicity, Immunologic; D000939:Epitopes / Q000276:immunology; D023061:Gene Order; D005818:Genetic Engineering; D005822:Genetic Vectors / Q000235:genetics; D015483:HIV Antibodies / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D015699:HIV Envelope Protein gp120 / Q000276:immunology; D015658:HIV Infections / Q000235:genetics / Q000276:immunology* / Q000628:therapy*; D015497:HIV-1 / Q000276:immunology*; D006801:Humans; D007167:Immunotherapy; D008213:Lymphocyte Activation / Q000276:immunology; D016131:Lymphocyte Subsets / Q000276:immunology* / Q000378:metabolism*; D011948:Receptors, Antigen, T-Cell / Q000235:genetics / Q000378:metabolism*; D019713:Receptors, CCR5 / Q000235:genetics / Q000378:metabolism; D011993:Recombinant Fusion Proteins; D057127:Single-Chain Antibodies; D014779:Virus Replication
chemical_list: D057134:Antibodies, Neutralizing; D000939:Epitopes; D015483:HIV Antibodies; D015699:HIV Envelope Protein gp120; D011948:Receptors, Antigen, T-Cell; D019713:Receptors, CCR5; D011993:Recombinant Fusion Proteins; D057127:Single-Chain Antibodies
keywords: CAR T cell; HIV; cell therapy; chimeric antigen receptor; gene editing