Coreceptor shift occurs in approximately half of all HIV infected individuals, and is promoted by use of CCR5 antagonist therapy. The proposed work uses a unique panel of CCR5/CXCR4 chimeric coreceptors and a panel of coreceptor shift HIV-1 isolates to examine the critical domains of CXCR4 required for HIV entry. The methodology includes ultra dense sequence analysis of HIV-1 gp120 sequences of quasispecies from U87.CD4 cells infected with env amplicons from a panel of dual/mixed isolates selected during CCR5 antagonist therapy in patient-specific virus libraries.
Aim 1. What proportion of HIV-1 gp120 sequences are responsible for VCVr resistance and how do HIV- 1 gp120 sequences of VCVs and VCVr isolates differ? For this purpose, U87.CD4.R5.X4 cells will be infected with patient-specific virus libraries in the presence or absence of VCV, and infected cell DNA subjected to ultra dense sequence analysis.
Aim 2. What proportion of HIV-1 gp120 sequences utilize CXCR4 for entry and how to the gp120 sequences differ from those that can not utilize CXCR4? For this purpose, U87.CD4.R5 and U87.CD4.X4 cells will be infected with patient-specific virus libraries, and infected cell DNA subjected to ultra dense sequence analysis.
Aim 3. What are the minimal domains of CXCR4 utilized for infection and how do HIV-1 gp120 sequences utilizing ECL3 or ECL2 differ from those that do not utilize these domains? For this purpose, U87.CD4 cells expressing chimeric CCR5/CXCR4 coreceptors will be infected with patient-specific virus libraries, and infected cell DNA subjected to ultra dense sequence analysis. Molecular details on the use of minimal domains of CXCR4 required for HIV entry will provide critical information to guide the development of more effective antiviral agents.

Public Health Relevance

HIV-1 isolates use a receptor, CD4, and a coreceptor, CCR5 and/or CXCR4 for entry into cells. Coreceptor shift occurs in approximately half of all HIV infected individuals, and is promoted by use of CCR5 antagonist therapy. The proposed work uses a unique panel of CCR5/CXCR4 chimeric coreceptors, and a panel of coreceptor shift HIV-1 isolates to examine the critical domains of CXCR4 required for HIV entry. What are the minimal domains of CXCR4 used by vicriviroc-resistant HIV-1 strains? To answer this question we will use envelope sequences, which represent a combination of multiple different envelope variants, from individual patients after acquisition of resistance to CCR5 antagonist, vicriviroc. Envelope sequences in each patient which promote entry into CXCR4 expressing cells will be determined by high resolution sequence analysis of infected cell DNA. A similar approach will be used to examine envelope sequences which allow entry into cells expressing specific domains of CXCR4, in order to determine the minimal domains required for HIV infection. Molecular details on the use of minimal domains of CXCR4 for HIV entry will provide critical information to guide the development of more effective antiviral agents.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
5R01AI106361-02
Application #
8631037
Study Section
AIDS Molecular and Cellular Biology Study Section (AMCB)
Program Officer
Sanders, Brigitte E
Project Start
2013-03-15
Project End
2015-02-28
Budget Start
2014-03-01
Budget End
2015-02-28
Support Year
2
Fiscal Year
2014
Total Cost
$190,000
Indirect Cost
$65,000
Name
Washington University
Department
None
Type
Schools of Medicine
DUNS #
068552207
City
Saint Louis
State
MO
Country
United States
Zip Code
63130