HIV/AIDS is a worldwide problem affecting 33 million people, with over 2 million deaths per year. Management of AIDS involves combinations of three or more antiretroviral drugs, but because HIV has the ability to mutate, new drug-resistant strains may be produced, rendering the treatment ineffective. Vaccines are currently designed and tested against prototype viruses that may not be representative of the diversity of viral quasispecies, and are inadequate for evaluation of the test vaccines. The study of viral proteins is mostly limited to single clones and unable to detect the contribution of genomic variability to the overall virus phenotype. This method can quickly generate recombinants that are representative of the viral genetic variability present in vivo, and thus can be a better tool for studying viral protein function.
Solution
A method to obtain viral recombinants that more effectively represents the quasispecies population has been developed. This approach is more rapid, efficient, and generates greater numbers of viral quasispecies than is currently attainable.
Competitive Advantage
More effective vaccines and therapeutic agents for pathogenic viruses can be rapidly developed using this approach. Furthermore, the method is a vast improvement over the existing ones in generating a greater number of viral recombinants quickly and efficiently.
Applications
Vaccine responses, drug susceptibility and viral fitness can be studied in infected individuals over time and among different individuals. This would be an extremely useful tool in developing antiretrovirals and/or HIV vaccines. In addition, the method could be used for the development of vaccines and therapeutic agents for other diseases caused by pathogenic viruses, including hepatitis, polio, and influenza viruses.
Patent Status
International Patent Application No. PCT/US2009/039660 was filed on April 6, 2009.
Licensing Opportunity
We are seeking commercialization partnerships for a novel, fast and efficient method for development of vaccines and therapeutic agents.
About the Inventors
Richard S. Myers has been cloning genes since the 1970s. He attended Iowa State University (1979-1983) specifically to work on plant biotechnology. His graduate work at the University of Illinois with Stanley Maloy focused on genetic approaches to dissect molecular machines in a bacterial pathogen, but he evaluated their successes by creating novel methods for enzyme studies including developing the first commercially offered software for enzyme kinetic analysis. After catching the recombination bug as a graduate student, Dr. Myers joined the lab of Frank Stahl (famous for his pivotal work in the most beautiful experiment in all of biology, the Meselson-Stahl experiment) where he contributed to toppling a famous recombination model and developed a love for viral recombination machines. He took up a position as a Fellow of the Canadian Institute of Advanced Research where he developed tools for functional genomics in the first sequenced bacterium (a pathogen). He also developed an interest in evolution and wrote an influential paper about the evolution of recombination hotspot dynamics. Since Dr. Myers moved to the University of Miami on Ground Hog's Day in 1997, he has been leading a research program devoted to untangling recombination mechanisms and putting them to work to create new opportunities for genetic engineers. When he isn't in the lab, Dr. Myers runs the Undergraduate Program in Biochemistry and Molecular Biology and is the Director for Undergraduate Research in BMB.
Rebeca Geffin is a Research Assistant Professor at the University of Miami Department of Pediatrics, in the Division of Infectious Diseases and Immunology. She arrived to the University as a post doctoral fellow in 1984 to work with a recently identified virus causing immunodeficiency in humans. Over the past 24 years, she has worked, followed the progress and the new discoveries in the field of HIV so that she not only possesses a wealth of knowledge in the subject, but also has a unique perspective that allows her to critically evaluate new ideas that have the potential for successful implementation. She has dedicated her scientific life to the study of infectious diseases, first at The Hebrew University of Jerusalem, where she obtained a Masters degree at the Department of Biochemistry, on the mechanism of entry of Sendai virus. Later she spent half a year at Yale University, where she worked in the laboratory of Nobel Laureate Paul Greengard. Her Doctorate was also obtained at the Hebrew University of Jerusalem on a lymphocyte derived protein that inhibited invasion of the malarial parasite Plasmodium falciparum. In Miami, she has focused her research effort on the study of immune and viral correlates of disease progression in children infected with HIV. She has authored original peer reviewed publications and has been recognized and cited by others. Her unique position as a basic scientist in a clinical department has given her exposure to the clinical aspects of HIV disease and has directed her research efforts to areas where discoveries could have a more direct relevance to modulation of disease.
Selected References
Myers RS, Stahl FW. ￇ and the RecBCD enzyme of Escherichia coli. Ann. Rev. Genet. 28:49-70, 1994.
Myers RS, Kuzminov A, Stahl FW. The recombination hotspot ￇ activates RecBCD recombination by converting E. coli to a recD mutant phenocopy. PNAS USA. 92:6244-6248, 1995.
Myers RS, Stahl MM, Stahl FW. ￇ recombination hotspot activity in phage ﾻ decays as a function of genetic distance. Genetics 141:805-812, 1995.
Boulton A, Myers RS, Redfield RJ. Evolution of recombination: The hotspot conversion paradox. PNAS USA. 94:8058-8063, 1997.
Jockovich ME, Myers RS. Nuclease activity is essential for RecBCD recombination in Escherichia coli. Molecular Microbiology 41:949-962, 2001.
Subramanian K, Rutvisuttinunt W, Scott W, Myers RS. The enzymatic basis of processivity in lambda exonuclease. Nucleic Acids Res. 31:1585-1596, 2003.
Vellani TS, Myers RS. Bacteriophage SPP1 Chu is an alkaline exonuclease in the Red family of viral two-component recombinases. J. Bacteriol. 185:2465-2474, 2003.
Reuven NB, Staire AE, Myers RS, Weller SK. The Herpes Simplex Virus-1 Alkaline Nuclease and Single-strand DNA binding Protein Mediate Strand Exchange in vitro. J. Virol. 77:7425-7433, 2003.
Thomason LC, Myers RS, Oppenheim AB, Costantino N, Sawitzke JA, Datta S, Bubunenko M, Court DL. Recombineering in Prokaryotes. In: Phages: Their Role in Bacterial Pathogenesis and Biotechnology; Ed. M.W. Waldor, D.I. Friedman, S.L. Adhaya, ASM Press; p. 383-399, 2005.
Larrea AA, Vaidyanathan PP, Malhotra A, Myers RS. Toward a Universal Mechanism for Mismatch Repair. Mol Cell. SUBMITTED (2008)
Geffin RB, Lai S, Hutto C, Scott G, Scott W, Master M, Parks WP. Quantitative analysis of human immunodeficiency virus type 1 antibody reactivity by Western Immunoblots. Evaluation of relative antibody levels in seropositive individuals and mothers. J. Inf. Dis. 165:111 118, 1992.
Schiller P, Geffin R, Voellmy R. Rapid complementation assay for anti HIV 1 drug screening and analysis of envelope protein function. AIDS Res. Hum. Retr. 8:1723 1731, 1992.
Hutto, C, Zhou Y, He J, Geffin R, Hill M, Scott W, Wood C. Longitudinal studies of viral sequence, viral phenotype and immunologic parameters in a pair of HIV-1 perinatally infected twins with discordant disease courses. J. Virol. 70:3589-3598, 1996.
Geffin RB, Scott GB, Melenwick M, Hutto C, Lai S, Boot LJ, McKenna P, Kessler JA, Conley AJ. Association of antibody reactivity to ELDKWA, a gp4l neutralization epitope, with disease progression in children perinatally infected with HIV-1. AIDS Res. Human Retr. 14:579-590, 1998.
Geffin RB, Wolf D, Muller R, Hill M, Stellwag E, Freitag M, Sass G, Scott G, Baur AS. Functional and structural defects in HIV type 1 genes derived from pediatric long-term survivors. AIDS Res. And Human Retr. 16:1855-1868, 2000.
Fackler OT, Wolf D, Weber HO, Laffert B, D'Aloja P, Schuler-Thurner B, Geffin R, Saksela K, Geyer M, Peterlin M, Schuler G, Baur AS. A natural variability in the proline-rich motif of Nef modulates HIV-1 replication in primary T cells. Curr. Biol 11:1294-9, 2001.
McCarthy M, He J., Auger D, Geffin R, Woodson C, Hutto, Wood C, Scott G. Cellular Tropisms and Co-receptor Usage of HIV-1 Isolates from Vertically Infected Children with Neurological Abnormalities and Rapid Disease Progression. J. Med. Virol. 67:1-8, 2002.
Kraus G, Geffin R, Spruill G, Young AK, Burzawa J, Cardona D, Hnatyszyn HJ. Cross-Clade Inhibition of HIV-1 Replication and Cytopathology Using Sequence-Specific RNase P Molecules. Proceedings of the National Academy of Sciences (PNAS). 99:3406-3411, 2002.
Geffin R, Hutto C, Andrew C, Scott GB. A Longitudinal Assessment of Autologous Neutralizing Antibodies in Children Perinatally Infected with HIV-1. Virology. 310:207-215, 2003.
McCarthy M, Vidaurre I, Geffin R. Maturing neurons are selectively sensitive to human immunodeficiency virus type 1 exposure in differentiating human neuroepithelial progenitor cell cultures. J. Neurovirol. 333:48, 2006.
