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Yukihiro Nishiyama MD, PhDProfessor, Department of Molecular Virology, Nagoya University Graduate School of Medicine

Specialized field


Career Summary

Nagoya University School of Medicine, M. D.
Nagoya University, Graduate School of Medicine, Ph. D.
Research Associate, Nagoya University School of Medicine.
Postdoctoral fellow, The Pennsylvania State University, Hershey, USA
Associate Professor, Research institute for Disease Mechanism and Control, Nagoya University School of Medicine
Professor, Laboratory of Virology, Research Institute for Disease Mechanism and Control, Nagoya University School of Medicine
Professor, Department of Virology, Nagoya University Graduate School of Medicine.

Research Theme

Fundamental and clinical research for cancer treatment using oncolytic herpes viruses

Research Summary

1. Development of oncolytic virotherapy

  Oncolytic viruses capable of tumor-selective replication and cytolysis have shown early promise as cancer therapeutics. We have developed replication-competent, attenuated herpes simplex virus-1 (HSV-1) mutants, named HF10 and Hh101, which have been evaluated for their oncolytic activities. However, the host immune system remains a significant obstacle to effective intraperitoneal administration of these viruses in the clinical setting. In this study, we investigated the use of these HSV-1 mutants as oncolytic agents against ovarian cancer and the use of human peritoneal mesothelial cells (MCs) as carrier cells for intraperitoneal therapy. MCs were efficiently infected with HSV-1 mutants, and MCs loaded with HSV-1 mutants caused cell killing adequately when cocultured with cancer cells in the presence or absence of HSV antibodies. In a mouse xenograft model of ovarian cancer, the injection of infected carrier cells led to a significant reduction of tumor volume and prolonged survival in comparison with the injection of virus alone. Our results indicate that replication-competent attenuated HSV-1 exerts a potent oncolytic effect on ovarian cancer, which may be further enhanced by the utilization of a carrier cell delivery system, based on amplification of viral load and possibly on avoidance of neutralizing antibodies.

2. Studies on the function of HSV accessory genes.

 Approximately half of the 74 genes encoded by HSV are accessory genes that are not essential for viral replication in cell-culture system . UL56 gene is an accessory gene encoded by most members of the Alphaherpesvirinae family. We have shown that UL56 facilitates the cytoplasmic transport of virions from the TGN to the plasma membrane and/or the release of virions. In addition, UL56 interacts with two other proteins: KIF1A , the neuron-specific kinesin; and HSV-2 UL11, a tegument protein that has dynamic membrane-trafficking properties and plays a role in the envelopment and egress of viral nucleocapsids . These interactions also support the view that UL56 is involved in transports of vesicles and virions, however the precise roles and functions of UL56 remain elusive. Our previous study has also shown that Itch, a Nedd4-family ligase, was identified as a UL56-interacting protein by a yeast two-hybrid screen. Itch is widely expressed in mammalian tissues, and Itch-deficient mice (itchy mice) develop a systemic and progressive autoimmune disease that proves lethal beginning at 6 months of age. Itch targets numerous proteins and has been implicated in signal transduction, endocytosis, differentiation, and transcription. In order to further elucidate the biological function of UL56, we examined the interaction of UL56 with the Nedd4-family ubiquitin ligase Itch and its role in the regulation of Itch. We found that, UL56 interacted with Itch, independent of additional viral proteins. Moreover, UL56 targeted Itch for degradation via both the lysosome and proteasome pathways. Compared to Nedd4, UL56 mediated more striking degradation of Itch. Ndfip and UL56 showed similar subcellular distribution and colocalized. UL56 did effect viral growth or decrease the expression of substrates for Itch. We believe that this is the first report demonstrating the interaction of a HSV-specific protein and Itch.

The UL14 protein of herpes simplex virus type 1 (HSV-1) is a minor tegument protein, expressed late in infection. VP16 (also known as Vmw65, α-trans-inducing factor, UL48 protein or ICP25) is the most abundant component of the tegument. It is well known that the virion-asoociated VP16 translocates into the nucleus by forming a complex with host cell factor 1 (HCF-1) and stimulates transcription of immediate early (IE) genes from the infecting genome. We previously constructed UL14-null (14D-VP16G) and rescued (14R-VP16G) viruses that expressed a VP16 GFP fusion protein, and have shown that UL14 protein enhances VP16 nuclear localization at the immediate eraly phase of infection, thus indirectly regulating the expression of IE genes. Here we report that a mutant protein VP16(Y364A) lacking HCF-1-binding ability was efficiently translocated into the nucleus by co-expressing UL14, suggesting that UL14 is more important for nuclear-translocation of VP16 than HCF-1. Moreover, we found that a major part of newly synthesized VP16-GFP accumulated in the perinuclear region of 14D-VP16G infected HEp-2 cells. The aggresome-like sturucture was surrounded by vimentin and Hsp70. Our data implicate that UL14 plays important roles in compartmentalization and/or folding-assist of VP16 during the late phase of infection.

Principal Research Achievement

  1. Iwata S, Wada K, Tobita S, Gotoh K, Ito Y, Demachi-Okamura A, Shimizu N, Nishiyama Y, Kimura H. Quantitative analysis of Epstein-Barr virus (EBV)-related gene expression in patients with chronic active EBV infection. J Gen Virol, 91, 42-50 (2010)
  2. Ishida D, Nawa A, Tanino T, Goshima F, Luo C. H, Iwaki M, Kajiyama H, Shibata K, Yamamoto E, Ino K, Tsurumi T, Nishiyama Y, Kikkawa F. Enhanced cytotoxicity with a novel system combining the paclitaxel-2’-ethylcarbonate produrug and an HSV amplicon with attenuated replication-competent virus, HF10 as a helper virus. Cancer Lett, 288, 17-27 (2010)
  3. Sato Y, Shirata N, Kudoh A, Iwahori S, Nakayama S, Murata T, Isomura H, Nishiyama Y, Tsurumi T. Expression of Epstein-Barr virus BZLF1 immediate-early protein induces p53 degradation independent of MDM2, leading to repression of p53-mediated transcription. Virology, 388, 204-211 (2009)
  4. Sato Y, Kamura T, Shirata N, Murata T, Kudoh A, Iwahori S, Nakayama S, Isomura H, Nishiyama, Y, Tsurumi T. Degradation of phosphorylated p53 by viral protein-ECS E3 ligase complex. PLo S Pathog, 7, e1000530 (2009)
  5. Kimura H, Miyake K, Yamauchi Y, Nishiyama K, Iwata S, Iwatsuki K, Gotoh K, Kojima S, Ito Y, Nishiyama Y. Identification of Epstein-Barr virus-infected lymphocyre subtypes by flow cytometric in situ hybridization in EBV-associated lymphoproliferative diseases. J Infect Dis, 200, 1078-1087 (2009)
  6. Ushijima Y, Goshima F, Kimura H, Nishiyama Y. Herpes simplex virus type 2 tegument protein UL56 relocalizes ubiquitin ligase Nedd4 and has a role in transport and/or release of virions. Virol J, 6, 168 (2009)
  7. Morimoto T, Arii J, Tanaka M, Sata T, Akashi H, Yamada M, Nishiyama Y. Uema M, Kawaguchi Y. Differences in the regulatory and functional effects of the US3 protein kinase activities of herpes simplex virus 1 and 2. J Virol, 83, 11624-11634 (2009)
  8. Ishikawa T, Yamada H, Oyamada A, Goshima F, Nishiyama Y, Yoshikai Y. Protective role of Fas-FasL signaling in lethal infection with herpes simplex virus type 2 in mice. J Virol, 83, 11777-11783 (2009)
  9. Yamauchi Y, Kiriyama K, Kubota N, Kimura H, Usukura J, Nishiyama Y. The UL14 tegument protein of herpes simplex virus type 1 is required for efficient nuclear transport of the alpha transinducing factor VP16 and viral capsids. J Virol, 82, 1094-1106 (2008)
  10. Watanabe D, Goshima F, Mori I, Tamada Y, Matsumoto Y, Nishiyama Y. Oncolytic viral therapy for malignant melanoma with herpes simplex virus type 1 mutant HF10. J Dermatol Sci, 50, 185-196 (2008)
  11. Gotoh K, Ito Y, Shibta-Watanabe Y, Kawada J, Takahashi Y, Yagasaki H, Kojima S, Nishiyama Y, Kimura H. Clinical and virologic characteristics of 15 patients with chronic active Epstein-Barr virus infection treated with hematopoietic stem cell transplantation. Clin Infect Dis, 46, 1525-1534 (2008)
  12. Nawa A, Luo C, Zhang L, Ushjima Y, Ishida D, Kamakura M, Fujimoto Y, Goshima F, Kikkawa F, Nishiyama Y. Non-engineered, naturally oncolytic herpes simplex virus HSV1 HF-10: applications for cancer gene therapy. Curr Gene Ther, 8, 208-221 (2008)
  13. Kato A, Tanaka M, Yamamoto M, Asai R, Sata T, Nishiyama Y, Kawaguchi Y. Identification of a physiological phosphorylation site of the Herpes simplex virus 1-encoded protein kinase Us3 which regulates its optimal catalytic activity in vitro and influences its function in infected cells. J Virol, 82, 6172-6189 (2008)
  14. Ushijima Y, Koshizuka T, Goshima F, Kimura H, and Nishiyama Y. Herpes Simplex Virus Type 2 UL56 Interacts with the Ubiquitin Ligase Nedd4 and Increases its Ubiquitination. J Virol, 82, 5220-5233 (2008)
  15. Yamauchi Y, Kiriyama K, Kimura H, Nishiyama, Y. Herpes Simplex Virus Induces Extensive Modification and Dynamic Relocalisation of the Nuclear Mitotic Apparatus Protein (NuMA) in Interphase Cells. J Cell Sci, 121, 2087-2096 (2008)
  16. Kamakura M et al. Microarray analysis of transcriptional responses to infection by herpes simplex virus types 1 and 2 and their US3-deficient mutants. Microb Infect 10: 405-413 (2008)
  17. Kohno S et al. Oncolytic Virotherapy with an HSV amplicon vector expressing granulocyte-macrophage colony-stimulating factor using the replication-competent HSV type I mutant HF10 as a helper virus. Cancer Gene Ther 14: 918-926 (2007)
  18. Kasuya H et al. Suitability of a US3-inactivated HSV mutant (L1BR1) as an oncolytic-virus for pancreatic cancer therpay. Cancer Gene Ther 14: 533-542 (2007)
  19. Kimata H et al. Pilot study of oncolytic viral therapy using mutant herpes simplex virus HF10 against recurrent metastatic breast cancer. Ann Surg Oncol 13: 1078-1084 (2006)
  20. Kato A et al. Herpes simplex virus 1-encoded protein kinase UL13 phosphorylates the viral US3 protein kinase and regulates nuclear localization of viral envelopment factors UL34 and UL31. J Virol. 80: 1476-1486 (2006)
  21. Nozawa N et al. Herpes simplex virus type 1 UL51 protein is involved in maturation and egress of virus particles. J Virol 79: 6947-6956 (2005)
  22. Kato A et al. Identification of proteins phosphorylated directly by the US3 protein kinase encoded by herpes simplex virus 1. J Virol 79: 9325-9331 (2005)
  23. Kudoh A et al. Inhibition of S-phase cyclin-dependent kinase activity blocks expression of Epstein Barr virus immediate early and early genes, preventing viral lytic replication. J Virol 78:104-115 (2004)
  24. Nishiyama Y et al. Herpes simplex virus gene products: the accessories reflect her lifestyle well. Rev Med Virol 14: 33-46 (2004)
  25. Kanamori M et al. Epstein-Barr virus nuclear antigen leader protein induces expression of thymus and activation-regulated chemokine in B cells. J Virol 78: 3984-3993 (2004)
  26. Nishimura H et al. Intraepithelial γδ T may bridge a gap between innate and acquired immunity to herpes simplex virus type 2. J Virol 78: 4927-4930 (2004)
  27. Kawaguchi Y et al. Conserved protein kinases encoded by herpesviruses and a cellular protein kinase cdc2 target the same phosphorylation site in eukaryotic elongation factor 1δ. J Virol 77: 2359-2368 (2003)
  28. Nozawa N et al. Subcellular localization of UL51 protein of herpes simplex virus type 1 and role of palmitoylation in Golgi targeting. J Virol 77: 3204-3216 (2003)
  29. Ihira M et al. Variation of human herpesvirus 7 shedding in saliva. J Infect Dis 188: 1352-1354 (2003)
  30. Yoshikawa T et al. Human herpesvirus 6 viremia in bone marrow transplant recipients:clinical features and risk factors. J Infect Dis 185: 847-853 (2002)
  31. Koshizuka T et al. Identification and characterization of the UL56 gene product of herpes simplex virus type 2. J Virol 76: 6718-6728 (2002)
  32. Yamauchi Y et al. Herpes simplex virus type 2 UL14 gene product has heat shock protein (HSP)-like functions. J Cell Sci 115: 2517-2527 (2002)
  33. Nishiyama Y et al. Antiapoptotic protein kinase of herpes simplex virus. Trend Microbiol 10:105-107 (2002)


Kojima Saburo Memorial Award for Distinguished Research on Infectious Diseases (2006).
Mochida Memorial Award for Medicine and Pharmaceutical Research (2006).