Educational Level:

Roadmap Objectives:

• • Article: Resistance to HIV-1 infection in Caucasian individuals bearing mutant alleles of the CCR-5 chemokine receptor gene. Samson et al. Nature, 1996; 382:722-5.
  • Content area/major concepts: I chose this paper as the starting point of the module because it identifies the gene responsible for HIV-1 resistance and includes population studies on the frequency of the mutant allele.
    
    HIV life cycle, receptor-mediated fusion, genetics, immunology, protein structure-function relationship
  • Methods or technology used to obtain data: PCR, DNA sequencing, mammalian cell culture, fusion assays, infection assays, genotyping
  • How the CREATE strategy was used:
  • Biggest teaching challenge: understanding diversity of HIV strains, need at least basic immunological knowledge, complexity of fusion experiment
• • Article: Maraviroc for previously treated patients with R5 HIV-1 infection. Gulick et al. NEJM, 2008; 359:1429-41
  • Content area/major concepts: I chose this paper to demonstrate how scientists used the knowledge of naturally occurring HIV resistance as the basis for new drug development. This paper also allows me to address how human clinical trials are carried out and analyzed.
    
    HIV fusion and pathology, receptor-ligand interactions, drug resistance
  • Methods or technology used to obtain data: Phase 3 Clinical Trial – includes studies on efficacy and safety (a LOT of stats!)
  • How the CREATE strategy was used:
  • Biggest teaching challenge: extensive statistical analysis with multiple variables and a large data set
• • Article: Long-term control of HIV by CCR5 delta32/delta32 stem-cell transplantation. Hütter et al. NEJM, 2009; 360:692-8.
  • Content area/major concepts: I chose this paper because they cured HIV!!! This paper describes the treatment of acute myeloid leukemia in a man with an HIV-1 infection. By purposefully choosing a stem cell donor that had the CCR5D32/D32 mutation, the researchers created an environment that should entirely inhibit viral replication by blocking fusion. This is the first proof of principle experiment that demonstrated that HIV could potentially be cured.
    
    Genetic variation, gene expression, receptor-ligand interactions, immunology, cancer, stem cells, HAART
  • Methods or technology used to obtain data: Genotyping, flow cytometry, chimerism analyses, ELISA, qPCR
  • How the CREATE strategy was used:
  • Biggest teaching challenge: Lack of knowledge about leukemia and stem-cell transplant therapy
• • Article: Effects of sequence changes in the HIV-1 gp41 fusion peptide on CCR5 inhibitor resistance. Anastassopoulou et al. Virology, 2012; 428:86-97.
  • Content area/major concepts: I chose this paper to begin to illustrate a molecular mechanism behind drug resistance. I think my students have several misconceptions and lack of knowledge about drug resistance. By the time they get to my class, I think that most of them understand that the drugs apply a selective pressure against microbes that are sensitive, which allows those that are resistant to grow. But, I don’t think they have any real idea of what the actual difference is between those that are sensitive and those that are resistant. This comes out in discussions when I try to emphasize that microbes that are resistant to one type of drug may not be resistant to other types.
    
    Protein-protein interactions, mutations and reversions, growth curves, drug resistance – how, why, varying levels
  • Methods or technology used to obtain data: Cell and viral culture, cloning and subcloning, infection assays, gene sequencing and sequence alignment
  • How the CREATE strategy was used:
  • Biggest teaching challenge: Keeping track of all of the different strains used and the shorthand notations used to identify them, understanding the big picture at the end – it would be very easy to get caught up in the altered protein-protein interactions and lose track of what that actually means in terms of the viral pathology
• • Article: Maraviroc and other HIV-1 entry inhibitors exhibit a class-specific redistribution effect that results in increased extracellular viral load. Kramer et al. Antimicrob Agents Chemother, 2012; epub ahead of printing.
  • Content area/major concepts: I chose this article because it addresses issues that may arise from using conventional testing methods (plasma viral load) as a detection method when using HIV fusion inhibitors instead of ARVs that act intracellularly. This will highlight the differences in the various classes of ARVs and get my students thinking more mechanistically about how drugs work (reinforcing some of the concepts from paper 4).
    
    Vertical gene transfer, HIV life cycle, HIV detection methods, drug mechanism of action
  • Methods or technology used to obtain data: Cell culture, transfection, spectrophotometry, enzyme (RT) activity assays
  • How the CREATE strategy was used:
  • Biggest teaching challenge: The techniques used in this paper are fairly straight forward and most have been encountered in previous papers in the module. The overall concept may be the biggest challenge – this drug increases viral load but is still an effective treatment – why?!

Advice for Using Module/Activity: