Dennis Hruby, Ph.D.
Vaccinia virus and anti-smallpox drugs
Research interests: Viral and bacterial pathogenesis,; virology, vaccines, bacterial virulence factors, anti-invectives
Office: 432 Nash Hall
Courses taught: MB 430/530 Pathogenic Microbiology, MB 434/534 Virology, MCB 525 Bacterial Expression Vectors, MB 636 Molecular Virology
Degrees: Ph.D., University of Colorado
RESEARCH
Our laboratory studies the interactions that occur between invading microbes and their target host cells which result in the induction of pathological consequences, or disease. We approach the study of microbial pathogenesis using two different model systems, on viral and the other bacterial. Our virology model is the mammalian poxvirus, vaccinia virus (VV). We are using molecular biological approaches to dissect the VV replicative cycle. In particular we are examining the role that posttranslational modifications such as acylation and proteolysis play in determining how viral proteins acquire functional integrity and become localized to the proper intracellular location in order to carry out their intended activity. Our bacterial model is the gram-positive bacteria, in particular the Group A streptococci. We are studying the pathways by which bacterial virulence factors, such as adhesins, become exported and anchored to the cell surface. Both projects share a common theme of basic research into a particular aspect of the host-pathogen relationship, followed by application of the derived knowledge to design prophylactic or therapeutic intervention strategies. Towards this goal, we are developing several recombinant vaccines and anti-infectives for potential use in humans. In recent years, we have focused on issues of biowarfare defense. To that end, we have developed a safer smallpox vaccine and two candidate smallpox antiviral drugs, all of which are about to enter animal testing en route towards clinical and regulatory approval.
RECENT and SELECTED PUBLICATIONS
Nalca A, Hatkin JM, Garza NL, Nichols DK, Norris SW, Hruby DE, Jordan R. Evaluation of orally delivered ST-246 as postexposure prophylactic and antiviral therapeutic in an aerosolized rabbitpox rabbit model. Antiviral Res. 2008 Aug;79(2):121-7. Epub 2008 Apr 23.
Jordan R, Tien D, Bolken TC, Jones KF, Tyavanagimatt SR, Strasser J, Frimm A, Corrado ML, Strome PG, Hruby DE. Single-dose safety and pharmacokinetics of ST-246, a novel orthopoxvirus egress inhibitor. Antimicrob Agents Chemother. 2008 May;52(5):1721-7. Epub 2008 Mar 3.
Duraffour S, Snoeck R, de Vos R, van Den Oord JJ, Crance JM, Garin D, Hruby DE, Jordan R, De Clercq E, Andrei G. Activity of the anti-orthopoxvirus compound ST-246 against vaccinia, cowpox and camelpox viruses in cell monolayers and organotypic raft cultures. Antivir Ther. 2007;12(8):1205-16.
Grosenbach DW, Jordan R, King DS, Berhanu A, Warren TK, Kirkwood-Watts DL, Tyavanagimatt S, Tan Y, Wilson RL, Jones KF, Hruby DE. Immune responses to the smallpox vaccine given in combination with ST-246, a small-molecule inhibitor of poxvirus dissemination. Vaccine. 2008 Feb 13;26(7):933-46. Epub 2007 Dec 26.
Berhanu A, Wilson RL, Kirkwood-Watts DL, King DS, Warren TK, Lund SA, Brown LL, Krupkin AK, Vandermay E, Weimers W, Honeychurch KM, Grosenbach DW, Jones KF, Hruby DE. Vaccination of BALB/c mice with Escherichia coli-expressed vaccinia virus proteins A27L, B5R, and D8L protects mice from lethal vaccinia virus challenge. J Virol. 2008 Apr;82(7):3517-29. Epub 2008 Jan 16.
Katritch V, Byrd CM, Tseitin V, Dai D, Raush E, Totrov M, Abagyan R, Jordan R, Hruby DE. Discovery of small molecule inhibitors of ubiquitin-like poxvirus proteinase I7L using homology modeling and covalent docking approaches. J Comput Aided Mol Des. 2007 Oct-Nov;21(10-11):549-58. Epub 2007 Oct 25.
Bolken TC, Hruby DE. Discovery and development of antiviral drugs for biodefense: experience of a small biotechnology company. Antiviral Res. 2008 Jan;77(1):1-5. Epub 2007 Aug 15. Review.
Quenelle DC, Prichard MN, Keith KA, Hruby DE, Jordan R, Painter GR, Robertson A, Kern ER. Synergistic efficacy of the combination of ST-246 with CMX001 against orthopoxviruses. Antimicrob Agents Chemother. 2007 Nov;51(11):4118-24. Epub 2007 Aug 27.
Yang SJ, Hruby DE. Vaccinia virus A12L protein and its AG/A proteolysis play an important role in viral morphogenic transition. Virol J. 2007 Jul 11;4:73.
Warren TK, Lund SA, Jones KF, Hruby DE. Comparison of transformation protocols in Streptococcus gordonii and evaluation of native promoter strength using a multiple-copy plasmid. Can J Microbiol. 2007 Mar;53(3):417-26.
Honeychurch KM, Yang G, Jordan R, Hruby DE. The vaccinia virus F13L YPPL motif is required for efficient release of extracellular enveloped virus. J Virol. 2007 Jul;81(13):7310-5. Epub 2007 May 2.
Sbrana E, Jordan R, Hruby DE, Mateo RI, Xiao SY, Siirin M, Newman PC, DA Rosa AP, Tesh RB. Efficacy of the antipoxvirus compound ST-246 for treatment of severe orthopoxvirus infection. Am J Trop Med Hyg. 2007 Apr;76(4):768-73.
Alzhanova D, Hruby DE. A host cell membrane protein, golgin-97, is essential for poxvirus morphogenesis. Virology. 2007 Jun 5;362(2):421-7. Epub 2007 Feb 5.
Quenelle DC, Buller RM, Parker S, Keith KA, Hruby DE, Jordan R, Kern ER. Efficacy of delayed treatment with ST-246 given orally against systemic orthopoxvirus infections in mice. Antimicrob Agents Chemother. 2007 Feb;51(2):689-95. Epub 2006 Nov 20.
Alzhanova D, Hruby DE. A trans-Golgi network resident protein, golgin-97, accumulates in viral factories and incorporates into virions during poxvirus infection. J Virol. 2006 Dec;80(23):11520-7. Epub 2006 Sep 20. Erratum in: J Virol. 2007 Feb;81(3):1539.
Moerdyk MJ, Byrd CM, Hruby DE. Analysis of vaccinia virus temperature-sensitive I7L mutants reveals two potential functional domains. Virol J. 2006 Aug 31;3:64.
Byrd CM, Hruby DE. Vaccinia virus proteolysis--a review. Rev Med Virol. 2006 May-Jun;16(3):187-202. Review.
Yoder JD, Chen TS, Gagnier CR, Vemulapalli S, Maier CS, Hruby DE. Pox proteomics: mass spectrometry analysis and identification of Vaccinia virion proteins. Virol J. 2006 Mar 1;3:10.
Honeychurch KM, Byrd CM, Hruby DE. Mutational analysis of the potential catalytic residues of the VV G1L metalloproteinase. Virol J. 2006 Feb 27;3:7.
Wilson RL, Brown LL, Kirkwood-Watts D, Warren TK, Lund SA, King DS, Jones KF, Hruby DE. Listeria monocytogenes 10403S HtrA is necessary for resistance to cellular stress and virulence. Infect Immun. 2006 Jan;74(1):765-8.
Bolken TC, Laquerre S, Zhang Y, Bailey TR, Pevear DC, Kickner SS, Sperzel LE, Jones KF, Warren TK, Amanda Lund S, Kirkwood-Watts DL, King DS, Shurtleff AC, Guttieri MC, Deng Y, Bleam M, Hruby DE. Identification and characterization of potent small molecule inhibitor of hemorrhagic fever New World arenaviruses. Antiviral Res. 2006 Feb;69(2):86-97. Epub 2005 Nov 28.
Yoder, J.D., T.S. Chen, C.R. Gagnier, S. Vemulapalli, C.S. Maier, and D.E. Hruby. (2006) Pox proteomics: mass spectrometry analysis and identification of Vaccinia virion proteins. Virology Journal. 3:10.
Honeychurch, K.M., C.M. Byrd, and D.E. Hruby. (2006) Mutational analysis of the potential catalytic residues of the VV G1L metalloproteinase. Virology Journal. 3:7.
Blouch, R.E., C.M. Byrd, and D.E. Hruby. (2005) Importance of disulphide bonds for vaccinia virus L1R protein function. Virology Journal. 2:91.
Byrd, C.M. and D.E. Hruby (2005) Development of an in vitro cleavage assay system to examine vaccinia virus I7L cysteine proteinase activity. Virology Journal 16:63.
Byrd, C.M., and D.E. Hruby (2005) A conditional-lethal vaccinia virus mutant demonstrates that the I7L gene product is required for virion morphogenesis. Virology Journal 2:4-9.
Warren, T. K., K. F. Jones and D. E. Hruby (2005) Use of a plasmid-borne expression (PLEX) system for the production of exogenous gene products by gram-positive bacterium Streptococcus gordonii. Protein Expression and Purification 40:319-326.
Byrd, C. M., T. C. Bolken, A. M. Mjalli, M. N. Arimilli, R. C. Andrews, R. Rothlein, T. Andrea, M. Rao, K. L. Owens and D. E. Hruby (2004) New Class of Orthopoxvirus Antiviral Drugs That Block Viral Maturation. Journal of Virology 78:12147-12156.
Olcott, M., C. Byrd, J. Watson and D.E. Hruby (2004) The VV G1L putative metalloproteinase is essential for virus replication in vivo. Journal of Virology 78:9947-9953.
Chen, T.S., J. D. Yoder, and D. E. Hruby (2004) Mass spectrometry analysis of synthetically myristylated peptides European Journal of Spectrometry 10:501-508.
Olcott, M. and D.E. Hruby (2004) Conditional Expression of vaccinia virus genes in mammalian cell lines expressing the tetracycline repressor. Journal of Virological Methods 120:9-12.
Yoder, J. D., T.S. Chen, and D. E. Hruby (2004) Sequence independent myristylation of a viral inclusion protein. Biochemistry 43:8297-8302.
Chen, T.S., J.D. Yoder, and D.E. Hruby (2003) Preparation of a large hydrophobic protein for mass spectrometry analysis: vaccinia virus ATI protein. Analytical Biochemistry . 315:277-280.
Byrd, C.M., T.C. Bolken, and D.E. Hruby (2002) The Vaccinia Virus I7L Gene Product is the Core Protein Proteinase. Journal of Virology . 76:8973-8976.
Hansen, S.G., T.A. Cope, and D.E. Hruby (2002) BiZyme: A Novel Fusion Protein-Mediating Selection of Vaccinia Virus Recombinants by Fluorescence and Antibiotic Resistance. BioTechniques . 32:1178-1187.
Byrd, C.M., T.C. Bolken, K.F. Jones, T.K. Warren, A.T. Vella, J. McDonald, D. King, Z. Blackwood, and D.E. Hruby. (2002) Biological Consequences of Antigen and Cytokine Co-Expression by Recombinant Streptococcus gordonii Vaccine Vectors. Vaccine . 20:2197-2205.
Geller, B.L., N. Wade, T.D. Gilberts, D.E. Hruby, R. Johanson, and L. Topisirovic (2001) Surface Expression of the Conserved C Repear Region of Streptococcal M6 Protein within the Pip Bacteriophage Receptor of Lactococcus lactis . Applied and Environmental Microbiology . 67(12): 5370-5376
Bolken, T.C., C.A. Franke, K.F. Jones, G.O. Zeller, E. Dutton, C.H. Jones, and D.E. Hruby (2001) Inactivation of the srtA gene in Streptococcus gordonii inhibits the cell wall anchoring of surface proteins and decreases in vitro and in vivo adhesion. Infection and Immunity 69:75-80.
Myscofski, D.M., E.K. Dutton, E. Cantor, A. Zhang, and D.E. Hruby (2001) Cleavage and Purification of Intein Fusion Protein Using the Streptococcus gordonii SPEX System. Prep. Biochem. & Biotechnol. 31:275-290.
Dutton, E.K., S. Ottum, C.A. Franke, and D.E. Hruby (2000) Expression of active monomeric and dimeric nuclease A from the Streptococcus gordonii surface protein expression (SPEX) system. Protein Purification and Expression 19:158-172.
Grosenbach, D.W., S.G. Hansen, and D.E. Hruby (2000) Identification and analysis of vaccinia virus palmitylproteins. Virology 275:193-206.
Myscofski, D.M., E. Dutton, T. Bolken, C. Franke, and D.E. Hruby (2000) Expression and purification of histidine tagged proteins from the gram-positive S. gordonii SPEX system. Protein Expression and Purification 20:112-123.
Franke, C., T. Bolman and D.E. Hruby (2000) Selection and characterization of a FUdR-resistant mutant of Streptococcus gordonii. Antimicrobial and Antiviral Chemotherapeutics 44:787-789