MOLECULAR BASIS OF PARVOVIRAL TARGET CELL SPECIFICITY

Summary

Principal Investigator: PETER J TATTERSALL
Abstract: DESCRIPTION (provided by applicant): This project uses genetic, biochemical and structural approaches to understand how autonomously replicating parvoviruses target and enter their host cells and establish infection. It will provide new information on the control of tissue tropis and on structural transitions in the virion that regulate successive steps in the infectious entry pathway of these ubiquitous viruses, particularly relating to an unexpected mode of genome exposure that could allow the capsid to fulfill novel intranuclear functions. The autonomously replicating parvoviruses are rugged and genetically simple single-stranded DNA viruses that are non-transforming because they are unable to activate resting cells to re-enter the cell cycle, and thus depend upon the host cell's regulation of cell cycle progression. In addition, many parvovirus species that infect rodents are inherently oncoselective, and preferentially infect transformed human cells, suggesting that they could be developed as therapeutic agents to target human tumors such as melanoma. The project pursues four approaches to understanding and exploiting such target specificity. 1. The targeting functions of the parvoviral shell will be identified by mapping the fortuitous tropism of parvovirus LuIII for human melanoma cells. This will involve making capsid chimeras with the non- melanotropic murine virus. Since LuIII is unlikely to be optimally melanotropic, we will use DNA shuffling to ask whether this property can be enhanced. 2. Differential qPCR analysis of sub-fractions of infected cells, expression of dominant-negative host genes and a novel in situ hybridization approach will be used to follow virions as they hijack specific intracellular trafficking pathways to gain access to the host nucleus. 3. Collaborative structural studies will be combined with genetic and biochemical approaches to investigate sequential conformational changes in the virion that facilitate ordered progress through its lifecycle. We have shown that genome uncoating is not a simple reversal of the packaging process, and will use these combined approaches to determine how the structurally symmetrical parvoviral capsid packages, retains and then releases its genome in the appropriate cellular compartments. 4. Optimized capsids will be used to package vectors designed to deliver heterologous genes into human and mouse melanoma cells. We will test the efficacy of a parvoviral vector packaged in an appropriately optimized capsid for targeting a transplantable murine melanoma in vivo. The vector expresses the B7-1 co-stimulatory molecule, and will potentially activate cytotoxic killer T-cells specific for melanoma tumor antigens, resulting in tumor eradication.
Funding Period: 1981-01-01 - 2017-04-30
more information: NIH RePORT

Top Publications

  1. pmc VP2 cleavage and the leucine ring at the base of the fivefold cylinder control pH-dependent externalization of both the VP1 N terminus and the genome of minute virus of mice
    Glen A Farr
    Department of Laboratory Medicine, Yale University Medical School, 333 Cedar Street, New Haven, CT 06510, USA
    J Virol 80:161-71. 2006
  2. pmc The family Parvoviridae
    Susan F Cotmore
    Department of Laboratory Medicine, Yale University School of Medicine, New Haven, CT, USA
    Arch Virol 159:1239-47. 2014
  3. pmc Distinct host cell fates for human malignant melanoma targeted by oncolytic rodent parvoviruses
    Ellen M Vollmers
    Medical Scientist Training Program, Yale University Medical School, 333 Cedar Street, New Haven, CT 06510, United States Department of Genetics, Yale University Medical School, 333 Cedar Street, New Haven, CT 06510, United States
    Virology 446:37-48. 2013
  4. pmc Parvoviral left-end hairpin ears are essential during infection for establishing a functional intranuclear transcription template and for efficient progeny genome encapsidation
    Lei Li
    Departments of Laboratory Medicine
    J Virol 87:10501-14. 2013
  5. pmc Parvovirus evades interferon-dependent viral control in primary mouse embryonic fibroblasts
    Lisa M Mattei
    Department of Immunobiology, Yale University, New Haven, CT 06520, USA
    Virology 442:20-7. 2013
  6. pmc Toll-like receptor 9 in plasmacytoid dendritic cells fails to detect parvoviruses
    Lisa M Mattei
    Department of Immunobiology, Yale University, New Haven, CT, USA
    J Virol 87:3605-8. 2013
  7. pmc Functional glycomic analysis of human milk glycans reveals the presence of virus receptors and embryonic stem cell biomarkers
    Ying Yu
    Department of Biochemistry and The Glycomics Center, Emory University School of Medicine, Atlanta, Georgia 30322, USA
    J Biol Chem 287:44784-99. 2012
  8. pmc LuIII parvovirus selectively and efficiently targets, replicates in, and kills human glioma cells
    Justin C Paglino
    Department of Neurosurgery, Yale University School of Medicine, New Haven, Connecticut, USA
    J Virol 86:7280-91. 2012
  9. pmc Mutations at the base of the icosahedral five-fold cylinders of minute virus of mice induce 3'-to-5' genome uncoating and critically impair entry functions
    Susan F Cotmore
    Department of Laboratory Medicine, Yale University School of Medicine, New Haven, Connecticut, USA
    J Virol 86:69-80. 2012
  10. pmc The parvoviral capsid controls an intracellular phase of infection essential for efficient killing of stepwise-transformed human fibroblasts
    Justin Paglino
    Department of Laboratory Medicine, Yale University Medical School, New Haven, CT 06520, USA
    Virology 416:32-41. 2011

Detail Information

Publications18

  1. pmc VP2 cleavage and the leucine ring at the base of the fivefold cylinder control pH-dependent externalization of both the VP1 N terminus and the genome of minute virus of mice
    Glen A Farr
    Department of Laboratory Medicine, Yale University Medical School, 333 Cedar Street, New Haven, CT 06510, USA
    J Virol 80:161-71. 2006
    ..5 or below. However, upon exposure to neutral pH following VP2 cleavage, its VP1-specific sequences and genome are extruded even at room temperature, underscoring the significance of the VP2 cleavage step for MVM particle dynamics...
  2. pmc The family Parvoviridae
    Susan F Cotmore
    Department of Laboratory Medicine, Yale University School of Medicine, New Haven, CT, USA
    Arch Virol 159:1239-47. 2014
    ..Also, affixes will be included in the names of genera to clarify subfamily affiliation and reduce the ambiguity that results from the vernacular use of "parvovirus" and "densovirus" to denote multiple taxon levels. ..
  3. pmc Distinct host cell fates for human malignant melanoma targeted by oncolytic rodent parvoviruses
    Ellen M Vollmers
    Medical Scientist Training Program, Yale University Medical School, 333 Cedar Street, New Haven, CT 06510, United States Department of Genetics, Yale University Medical School, 333 Cedar Street, New Haven, CT 06510, United States
    Virology 446:37-48. 2013
    ....
  4. pmc Parvoviral left-end hairpin ears are essential during infection for establishing a functional intranuclear transcription template and for efficient progeny genome encapsidation
    Lei Li
    Departments of Laboratory Medicine
    J Virol 87:10501-14. 2013
    ..These results suggest that the mutant virions reach the nucleus, uncoat, and are converted to duplex DNA but require an intact left-end hairpin structure to form the initiating transcription complex. ..
  5. pmc Parvovirus evades interferon-dependent viral control in primary mouse embryonic fibroblasts
    Lisa M Mattei
    Department of Immunobiology, Yale University, New Haven, CT 06520, USA
    Virology 442:20-7. 2013
    ..Together, these data suggest that the MVMp efficiently evades antiviral immune mechanisms imposed by type I IFNs, which may in part explain their efficient transmission between mice...
  6. pmc Toll-like receptor 9 in plasmacytoid dendritic cells fails to detect parvoviruses
    Lisa M Mattei
    Department of Immunobiology, Yale University, New Haven, CT, USA
    J Virol 87:3605-8. 2013
    ..Here we have shown that despite the ability of purified genomic DNA to stimulate TLR9 and despite the ability to enter TLR9 endosomes, ssDNA viruses of the Parvoviridae family failed to elicit an interferon (IFN) response in pDCs...
  7. pmc Functional glycomic analysis of human milk glycans reveals the presence of virus receptors and embryonic stem cell biomarkers
    Ying Yu
    Department of Biochemistry and The Glycomics Center, Emory University School of Medicine, Atlanta, Georgia 30322, USA
    J Biol Chem 287:44784-99. 2012
    ..Together, these results provide novel insights into diverse recognition functions of HMGs and show the utility of the SGM approach and MAGS as resources for defining novel glycan recognition by GBPs, antibodies, and pathogens...
  8. pmc LuIII parvovirus selectively and efficiently targets, replicates in, and kills human glioma cells
    Justin C Paglino
    Department of Neurosurgery, Yale University School of Medicine, New Haven, Connecticut, USA
    J Virol 86:7280-91. 2012
    ..Intravenous or intracranial LuIII caused no adverse effects. Intracranial LuIII caused no infection of mature mouse neurons or glia in vivo but showed a modest infection of developing neurons...
  9. pmc Mutations at the base of the icosahedral five-fold cylinders of minute virus of mice induce 3'-to-5' genome uncoating and critically impair entry functions
    Susan F Cotmore
    Department of Laboratory Medicine, Yale University School of Medicine, New Haven, Connecticut, USA
    J Virol 86:69-80. 2012
    ..However, unlike the wild type, the V40A mutant extensively uncoated during cell entry, indicating that the V40-L172 interaction restrains an uncoating trigger mechanism within the endosomal compartment...
  10. pmc The parvoviral capsid controls an intracellular phase of infection essential for efficient killing of stepwise-transformed human fibroblasts
    Justin Paglino
    Department of Laboratory Medicine, Yale University Medical School, New Haven, CT 06520, USA
    Virology 416:32-41. 2011
    ..Thus targeting of human cancers of different tissue-type origins will require use of parvoviruses with capsids that effectively make this critical interaction...
  11. pmc Structure of a packaging-defective mutant of minute virus of mice indicates that the genome is packaged via a pore at a 5-fold axis
    Pavel Plevka
    Department of Biological Sciences, Purdue University, West Lafayette, Indiana 47907, USA
    J Virol 85:4822-7. 2011
    ..The mutant capsid had its 5-fold channel blocked, and the particles were unable to package DNA, strongly suggesting that the 5-fold pore is the packaging portal for genome entry...
  12. pmc Recruitment of DNA replication and damage response proteins to viral replication centers during infection with NS2 mutants of Minute Virus of Mice (MVM)
    Zandra Ruiz
    Department of Laboratory Medicine, Yale University School of Medicine, 333 Cedar Street, New Haven, CT 06510, USA
    Virology 410:375-84. 2011
    ..We conclude that MVM infection invokes multiple damage responses that influence the APAR environment, but that NS2 does not modify the recruitment of cellular proteins...
  13. pmc Depletion of virion-associated divalent cations induces parvovirus minute virus of mice to eject its genome in a 3'-to-5' direction from an otherwise intact viral particle
    Susan F Cotmore
    Department of Laboratory Medicine, Yale University Medical School, 333 Cedar Street, New Haven, CT 06510, USA
    J Virol 84:1945-56. 2010
    ....
  14. pmc Evolution to pathogenicity of the parvovirus minute virus of mice in immunodeficient mice involves genetic heterogeneity at the capsid domain that determines tropism
    Alberto López-Bueno
    Centro de Biología Molecular Severo Ochoa Consejo Superior de Investigaciones Científicas Universidad Autónoma de Madrid, 28049 Cantoblanco, Madrid, Spain
    J Virol 82:1195-203. 2008
    ..The evolutionary changes delineate a sector of the surface of the capsid that determines tropism and that surrounds the sialic acid receptor binding domain...
  15. ncbi Parvoviral host range and cell entry mechanisms
    Susan F Cotmore
    Department of Laboratory Medicine, Yale University Medical School, New Haven, Connecticut 06510, USA
    Adv Virus Res 70:183-232. 2007
    ....
  16. pmc Exploring the contribution of distal P4 promoter elements to the oncoselectivity of Minute Virus of Mice
    Justin Paglino
    Department of Laboratory Medicine, Yale University Medical School, 333 Cedar Street, New Haven, CT 067510, USA
    Virology 361:174-84. 2007
    ..We also confirmed that reducing the PIF half-site spacing by one basepair enhances oncoselectivity, but found that a further basepair deletion significantly reduces this effect...
  17. ncbi Differential roles for the C-terminal hexapeptide domains of NS2 splice variants during MVM infection of murine cells
    Zandra Ruiz
    Graduate Program in Microbiology, Yale University, 333 Cedar Street, New Haven, CT 06510, USA
    Virology 349:382-95. 2006
    ..Thus, the NS2P isoform, even when expressed at a level lower than that of NS2Y, performs a critical role in infection of A9 cells that cannot be accomplished by the NS2Y isoform alone...
  18. pmc Profiling of glycan receptors for minute virus of mice in permissive cell lines towards understanding the mechanism of cell recognition
    Sujata Halder
    Department of Biochemistry and Molecular Biology, University of Florida, Gainesville, Florida, United States of America
    PLoS ONE 9:e86909. 2014
    ....

Research Grants30

  1. Novel Ad/MVA and Ad/Protein HIV-1 Vaccines
    Dan H Barouch; Fiscal Year: 2013
    ..To define the mechanism of blocking acquisition of stringent SIV challenges by conducting antigen formulation and adoptive transfer studies in rhesus monkeys. ..
  2. Cardiac Myosin Binding Protein-C: Structure, Function, and Regulation
    David M Warshaw; Fiscal Year: 2013
    ..abstract_text> ..
  3. Pacific NorthWest Regional Center of Excellence (PNWRCE)
    Jay A Nelson; Fiscal Year: 2013
    ..pseudomallei host pathogen response during both the septicemic as well as the intracellular phases of the disease. ..
  4. Southeast Regional Centers of Excellence for Biodefense &Emerging Infectious Di
    Philip Frederick Sparling; Fiscal Year: 2013
    ..SERCEB brings new investigators to the biodefense effort through a combination of educational programs, support of innovative new projects, and the synergistic interactions among its world-class investigators. ..
  5. Northeast Biodefense Center
    W Ian Lipkin; Fiscal Year: 2013
    ..As a Center based in a School of Public Health and a State Department of Health, the NBC has a firm commitment to and practical understanding of Emergency Preparedness. ..
  6. Pacific Southwest RCE for Biodefense &Emerging Infectious Diseases Research
    Alan G Barbour; Fiscal Year: 2013
    ..abstract_text> ..
  7. Functional Annotation of the Pancreatic Cancer Genome
    Steven D Leach; Fiscal Year: 2013
    ..Together, these studies will dramatically accelerate the functional annotation of the pancreatic cancer genome, setting the stage for future therapeutic applications. ..
  8. Rocky Mountain Regional Center of Excellence or Biodefense and Emerging Infectiou
    John T Belisle; Fiscal Year: 2013
    ..abstract_text> ..
  9. Directing Tumor-specific T cells to Tumors
    Pawel Kalinski; Fiscal Year: 2013
    ..abstract_text> ..
  10. UNMC EPPLEY CANCER CENTER SUPPORT GRANT
    Kenneth H Cowan; Fiscal Year: 2013
    ....