title: Targeting the HIV entry, assembly and release pathways for anti-HIV gene therapy.
abstract: Targeting the HIV entry and assembly pathways holds promise for development of novel anti-HIV gene therapy vectors. We characterized discrete dominant negative (DN) Gag and Envelope mutants for their anti-HIV-1 activity. We show here that capsid mutants (Q155N and Y164A) are more potent inhibitors of WT HIV than the matrix mutant 1GA. Both the Envelope mutants tested, V513E and R515A, were equally effective and a combination of Gag and Envelope DN genes significantly enhanced potency. Interestingly, the DN mutants acted at multiple steps in the virus life cycle rather than solely disrupting virus release or infection. Inhibition mediated by R515A could be partially attributed to the Envelope cytoplasmic tail, as deletion of R515A tail partially abrogated its DN effect. Finally, the Y164A/R515A double mutant expressed in a lentiviral vector was effective at inhibiting HIV replication in CD34+ hematopoietic stem cell-derived macrophages, demonstrating the therapeutic potential of our approach.
authors.affiliation: Center of Excellence for Infectious Disease, Department of Biomedical Sciences, Texas Tech University Health Sciences Center, El Paso, TX 79905, USA. anjali.joshi@ttuhsc.edu
authors.forename: Anjali
authors.initials: A
authors.lastname: Joshi
authors.forename: Himanshu
authors.initials: H
authors.lastname: Garg
authors.forename: Sherimay
authors.initials: S
authors.lastname: Ablan
authors.forename: Eric O
authors.initials: EO
authors.lastname: Freed
authors.forename: Kunio
authors.initials: K
authors.lastname: Nagashima
authors.forename: N
authors.initials: N
authors.lastname: Manjunath
authors.forename: Premlata
authors.initials: P
authors.lastname: Shankar
mesh_terms: D002460:Cell Line; D015658:HIV Infections / Q000188:drug therapy / Q000821:virology*; D015497:HIV-1 / Q000235:genetics* / Q000502:physiology; D006801:Humans; D009154:Mutation; D019065:Virus Assembly; D053586:Virus Internalization; D014779:Virus Replication; D017201:Virus Shedding; D054299:env Gene Products, Human Immunodeficiency Virus / Q000235:genetics* / Q000378:metabolism / Q000627:therapeutic use; D054301:gag Gene Products, Human Immunodeficiency Virus / Q000235:genetics* / Q000378:metabolism / Q000627:therapeutic use
chemical_list: D054299:env Gene Products, Human Immunodeficiency Virus; D054301:gag Gene Products, Human Immunodeficiency Virus