title: Evolution of coreceptor utilization to escape CCR5 antagonist therapy.
abstract: The HIV-1 envelope interacts with coreceptors CCR5 and CXCR4 in a dynamic, multi-step process, its molecular details not clearly delineated. Use of CCR5 antagonists results in tropism shift and therapeutic failure. Here we describe a novel approach using full-length patient-derived gp160 quasispecies libraries cloned into HIV-1 molecular clones, their separation based on phenotypic tropism in vitro, and deep sequencing of the resultant variants for structure-function analyses. Analysis of functionally validated envelope sequences from patients who failed CCR5 antagonist therapy revealed determinants strongly associated with coreceptor specificity, especially at the gp120-gp41 and gp41-gp41 interaction surfaces that invite future research on the roles of subunit interaction and envelope trimer stability in coreceptor usage. This study identifies important structure-function relationships in HIV-1 envelope, and demonstrates proof of concept for a new integrated analysis method that facilitates laboratory discovery of resistant mutants to aid in development of other therapeutic agents.
authors.affiliation: Division of Molecular Oncology, Department of Medicine, Washington University School of Medicine, St. Louis, MO, USA.
authors.forename: Jie
authors.initials: J
authors.lastname: Zhang
authors.affiliation: Division of Molecular Oncology, Department of Medicine, Washington University School of Medicine, St. Louis, MO, USA.
authors.forename: Xiang
authors.initials: X
authors.lastname: Gao
authors.affiliation: The McDonnelle Genome Institute, Washington University School of Medicine, St. Louis, MO, USA.
authors.forename: John
authors.initials: J
authors.lastname: Martin
authors.affiliation: The McDonnelle Genome Institute, Washington University School of Medicine, St. Louis, MO, USA.
authors.forename: Bruce
authors.initials: B
authors.lastname: Rosa
authors.affiliation: Department of Computer Science and Engineering, Washington University in St. Louis, St. Louis, MO, USA.
authors.forename: Zheng
authors.initials: Z
authors.lastname: Chen
authors.affiliation: The McDonnelle Genome Institute, Washington University School of Medicine, St. Louis, MO, USA.
authors.forename: Makedonka
authors.initials: M
authors.lastname: Mitreva
authors.affiliation: Division of Infectious Diseases, Brigham and Women׳s Hospital, Harvard Medical School, MA, USA.
authors.forename: Timothy
authors.initials: T
authors.lastname: Henrich
authors.affiliation: Division of Infectious Diseases, Brigham and Women׳s Hospital, Harvard Medical School, MA, USA.
authors.forename: Daniel
authors.initials: D
authors.lastname: Kuritzkes
authors.affiliation: Division of Molecular Oncology, Department of Medicine, Washington University School of Medicine, St. Louis, MO, USA. Electronic address: lratner@dom.wustl.edu.
authors.forename: Lee
authors.initials: L
authors.lastname: Ratner
mesh_terms: D000483:Alleles; D000595:Amino Acid Sequence; D019380:Anti-HIV Agents / Q000494:pharmacology* / Q000627:therapeutic use; D065100:CCR5 Receptor Antagonists / Q000494:pharmacology* / Q000627:therapeutic use; D002460:Cell Line; D019143:Evolution, Molecular; D005787:Gene Frequency; D015699:HIV Envelope Protein gp120 / Q000737:chemistry / Q000235:genetics; D015658:HIV Infections / Q000188:drug therapy / Q000235:genetics* / Q000378:metabolism / Q000821:virology*; D015497:HIV-1 / Q000145:classification / Q000187:drug effects* / Q000235:genetics*; D059014:High-Throughput Nucleotide Sequencing; D006801:Humans; D008958:Models, Molecular; D010446:Peptide Fragments / Q000737:chemistry / Q000235:genetics; D010802:Phylogeny; D020641:Polymorphism, Single Nucleotide; D056510:Position-Specific Scoring Matrices; D011487:Protein Conformation; D012367:RNA, Viral; D019713:Receptors, CCR5 / Q000235:genetics* / Q000378:metabolism; D019718:Receptors, CXCR4 / Q000235:genetics / Q000378:metabolism; D013329:Structure-Activity Relationship; D056189:Viral Tropism; D054299:env Gene Products, Human Immunodeficiency Virus / Q000737:chemistry / Q000235:genetics
chemical_list: D019380:Anti-HIV Agents; D065100:CCR5 Receptor Antagonists; D015699:HIV Envelope Protein gp120; C059762:HIV envelope protein gp120 (305-321); D010446:Peptide Fragments; D012367:RNA, Viral; D019713:Receptors, CCR5; D019718:Receptors, CXCR4; D054299:env Gene Products, Human Immunodeficiency Virus
keywords: Coreceptor inhibitor; Drug resistance; HIV/AIDS; Tropism shift