title: A highly efficient short hairpin RNA potently down-regulates CCR5 expression in systemic lymphoid organs in the hu-BLT mouse model.
abstract: Inhibiting the expression of the HIV-1 coreceptor CCR5 holds great promise for controlling HIV-1 infection in patients. Here we report stable knockdown of human CCR5 by a short hairpin RNA (shRNA) in a humanized bone marrow/liver/thymus (BLT) mouse model. We delivered a potent shRNA against CCR5 into human fetal liver-derived CD34(+) hematopoietic progenitor/stem cells (HPSCs) by lentiviral vector transduction. We transplanted vector-transduced HPSCs solidified with Matrigel and a thymus segment under the mouse kidney capsule. Vector-transduced autologous CD34(+) cells were subsequently injected in the irradiated mouse, intended to create systemic reconstitution. CCR5 expression was down-regulated in human T cells and monocytes/macrophages in systemic lymphoid tissues, including gut-associated lymphoid tissue, the major site of HIV-1 replication. The shRNA-mediated CCR5 knockdown had no apparent adverse effects on T-cell development as assessed by polyclonal T-cell receptor Vbeta family development and naive/memory T-cell differentiation. CCR5 knockdown in the secondary transplanted mice suggested the potential of long-term hematopoietic reconstitution by the shRNA-transduced HPSCs. CCR5 tropic HIV-1 infection was effectively inhibited in mouse-derived human splenocytes ex vivo. These results demonstrate that lentiviral vector delivery of shRNA into human HPSCs could stably down-regulate CCR5 in systemic lymphoid organs in vivo.
authors.affiliation: Department of Medicine, Division of Hematology-Oncology, UCLA AIDS Institute, David Geffen School of Medicine, University of California-Los Angeles, 615 Charles E. Young Drive South, Los Angeles, CA 90095, USA.
authors.forename: Saki
authors.initials: S
authors.lastname: Shimizu
authors.forename: Patrick
authors.initials: P
authors.lastname: Hong
authors.forename: Balamurugan
authors.initials: B
authors.lastname: Arumugam
authors.forename: Lauren
authors.initials: L
authors.lastname: Pokomo
authors.forename: Joshua
authors.initials: J
authors.lastname: Boyer
authors.forename: Naoya
authors.initials: N
authors.lastname: Koizumi
authors.forename: Panyamol
authors.initials: P
authors.lastname: Kittipongdaja
authors.forename: Angela
authors.initials: A
authors.lastname: Chen
authors.forename: Greg
authors.initials: G
authors.lastname: Bristol
authors.forename: Zoran
authors.initials: Z
authors.lastname: Galic
authors.forename: Jerome A
authors.initials: JA
authors.lastname: Zack
authors.forename: Otto
authors.initials: O
authors.lastname: Yang
authors.forename: Irvin S Y
authors.initials: IS
authors.lastname: Chen
authors.forename: Benhur
authors.initials: B
authors.lastname: Lee
authors.forename: Dong Sung
authors.initials: DS
authors.lastname: An
mesh_terms: D000818:Animals; D001853:Bone Marrow / Q000378:metabolism*; D002454:Cell Differentiation / Q000235:genetics; D004195:Disease Models, Animal; D015536:Down-Regulation; D055785:Gene Knockdown Techniques; D015658:HIV Infections / Q000235:genetics / Q000378:metabolism*; D015497:HIV-1; D018380:Hematopoietic Stem Cell Transplantation; D006412:Hematopoietic Stem Cells / Q000378:metabolism; D006801:Humans; D007156:Immunologic Memory / Q000235:genetics; D016086:Lentivirus; D008099:Liver / Q000378:metabolism*; D051379:Mice; D016688:Mice, Inbred NOD; D016693:Receptors, Antigen, T-Cell, alpha-beta / Q000378:metabolism; D019713:Receptors, CCR5 / Q000096:biosynthesis* / Q000235:genetics; D013601:T-Lymphocytes / Q000378:metabolism; D013950:Thymus Gland / Q000378:metabolism*; D014161:Transduction, Genetic; D014183:Transplantation, Heterologous
chemical_list: D016693:Receptors, Antigen, T-Cell, alpha-beta; D019713:Receptors, CCR5