Scientific Accomplishments

The Viral Oncology (VO) research program is one of OSUCCC–James' six highly interactive cancer Research Programs. The programs comprise more than 200 leading scientists from 13 different colleges within OSU and affiliated academic institutions.

VO promotes collaborative research in order to discover mechanisms of virus-related neoplastic disease, design improved viral vector-mediated gene transfer technologies and improve oncolytic virotherapy.

Accomplishments include:

Programmatic Funding 2009

  • Program Project Grant: The study of highly integrated retrovirus models to elucidate essential cellular mechanisms that define lymphocyte transformation and disease. – Michael Lairmore, DVM, PhD
  • NINDS Grant: U01NS061811, “Preclinical toxicity evaluation of a potent onoclytic virus” – E. Antonio Chiocca, MD, PhD
  • Grant (OSU 0455): “A Phase I Study of Adv-Tk + Valacyclovir Gene Therapy in Combination with Standard Radiation Therapy for Malignant Gliomas” – E. Antonio Chicca, MD, PhD
  • Grant (OSU 07059): “A Phase IIa Study of Adv-Tk + Valacyclovir Gene Therapy in Combination with Standard Radiation Therapy for Malignant Gliomas” – E. Antonio Chicca, MD, PhD
  • Grant (OSU 07059): “A Phase I/II Clinical Trial to Evaluate Dose Limiting Toxicity & Efficacy of Intralesional Administration of Reolysin for the Treatment of Patiens with Histologically Confirmed Recurrent Malignant Gliomas ” – E. Antonio Chicca, MD, PhD

Selected Findings & Developments

J Natl Cancer Inst 100(6): 407-20., 2008. Distinct risk factor profiles for human papillomavirus type 16-positive and human papillomavirus type 16-negative head and neck cancers (Gillison, M).
Key Finding: Identified oral sexual behavior as the principal risk factor for oral HPV infection and HPV-associated oral cancers.

J Natl Cancer Inst 99(23):1768-81, 2007 Effect of tumor microenvironment modulation on the efficacy of oncolytic virus therapy (Carson W, Chiocca EA, Kaur B) [Featured editorial]
Key Finding:  A fundamental limitation to the success of oncolytic virus therapy of gliomas is the hyperpermeability and inflammatory response of tumors.

PNAS  103(34):12873-8, 2006  Cyclophosphamide enhances glioma virotherapy by inhibiting innate immune responses (Caligiuri M, Chiocca EA, Kaur B)
Key Finding: Cyclophosphamide significantly enhances the anticancer effect of oncolytic viruses through reduction of innate immunity.

J Clin Oncol 23(27):6481-8, 2005 (Gross TG et al.) Low-dose chemotherapy for Epstein-Barr virus-positive post-transplantation lymphoproliferative disease in children after solid organ transplantation
Key Finding: Low-dose chemotherapy regimen is effective for children with EBV-positive, nonfulminant PTLD who have experienced treatment failure with front-line therapy. This study represents the largest series of PTLD patients treated prospectively with a uniform chemotherapy regimen.

Nat Struc Mol Biol 13(6):509-16., 2006 RNA helicase A is necessary for translation of selected messenger RNAs (Boris-Lawrie K, Schoenberg D, Fernandez S). Journal Cover Story.
Key Finding: Discovered a new regulatory axis of growth genes of cells and of viruses that is operated by a specific RNA-RNA helicase A interaction. RNA Helicase A is also a biomarker of human tumors.

Blood 112:3788-3797, 2008 HTLV-1 antisense encoded gene, Hbz, promotes T-lymphocyte proliferation (Green PL, Lairmore MD)
Key Finding: This work provided direct evidence that HTLV-1 Hbz, which has now been shown to be the only viral gene expressed in ATL cells plays a role in tumor cell survival and organ infiltration.  Hbz will be a key therapeutic target.

Recognition, Initiatives, Honors & Awards (2008 & 2009)

U01 Project Award

VO Co-Leader Antonio Chiocca, MD, PhD, was awarded a U01 project with VO member Michael Caligiuri, MD, and Viral Oncology Research Program member Michael Oglesbee, PhD. The objective: to perform the necessary preclinical toxicology studies to bring to clinical trials an OV based on HSV1 that targets for replication nestin-expressing glioma cells, likely to represent the glioma stem-like subpopulation that enables tumor maintenance and survival.

Program Progent Grant

P01CA1007030-07, VO member Michael Lairmore, DVM, PhD PI. This proposal continues the study of highly integrated retrovirus models to elucidate essential cellular mechanisms that define lymphocyte transformation and disease. The study of retroviruses has provided fundamental and critical knowledge for defining cell transformation mechanisms as well as fundamental cell biology paradigms. The VO will extend upon the established interactions through inter-related goals to define critical events that control lymphocyte transition from activation through immortalization and transformation.

Project 1 Dr. Lairmore uses retroviruses to provide novel insights into how they alter transcription and DNA damage/repair signaling to establish persistent infections and set the stage for cellular transformation.
Project 2 VO Co-Leader Patrick L. Green, PhD will define novel, post-transcriptional mechanisms of retroviruses and, working with Dr. Lairmore (Project 1), will comparatively test each protein in viral replication and lymphocyte transformation.
Project 3 VO member Kathleen Boris-Lawrie, PhD, working collaboratively with multiple PPG investigators will examine the fundamental translational control mechanisms in both retroviruses and in crucial growth control genes.
Project 4 Weilbaecher, PhD Washington University, and Molecular Carcinogenesis & Chemoprevention Research Program member Thomas Rosol, DVM, PhD, combine their expertise with PPG investigators to study the role of bone microenvironment in osteolytic and osteoblastic tumor models.
Project 5 Lee Ratner, PhD Washington University continues the highly dynamic collaborations between each university to test the contribution of the inflammatory microenvironment in carcinogenesis using unique transgenic mouse models.

The PPG comprises the five interactive projects above as well as three highly supportive Cores (Dr. Lairmore, Mathes, PhD, Fernandez, PhD (Statistics), Kvaraskhelia, PhD (ET), and  Niewiesk, PhD). 

Each project is related to two central components involving the regulation of lymphocyte signaling and gene expression by retroviruses or cellular microenvironment alterations leading to transformation and paraneoplastic disease. The PPG’s interactive nature will provide rational hypotheses to support translational studies to define therapeutic intervention against retrovirus-induced lymphoma and refine animal models to determine molecular determinants of retrovirus-induced cancer.

Discoveries & Inventions

CMV-Specific MicroRNA Signatures in Gliomas

VO members Sean Lawler, PhD, and Antonio Chiocca, MD, PhD, have discovered CMV-specific microRNA signatures in gliomas from human patients. The team is currently attempting to validate these signatures and discover their targets. If true, the research may provide the first causative links between this virus and malignant gliomas. The discovery is part of a pilot project led by Dr. Chiocca, Chang-hyuk Kwon, PhD, and Charles Cook, PhD which focuses on validating the hypothesis of an association between cytomegalovirus (CMV) and glioma genesis and progression.

PBM-Mediated Associations Between Tax-1 & Cellular Proteins

As a result of their studies of HTLV-1 transforming protein, Tax, its role in the virus-host interface and, ultimately, cellular transformation and disease induction, VO Co-Leader Patrick Green, PhD, and his team’s infection and proliferation assays revealed that the Tax-1 PBM significantly increases both HTLV-1 and HTLV-2-induced primary T-cell proliferation. Additionally, Tax-1 PBM was responsible for Tax-1’s micronuclei/DNA damage induction activity relative to that of Tax-2.  Viral infection and persistence were severely attenuated in rabbits inoculated with HTLV-1PBM.  These results offer the first direct evidence that PBM-mediated associations between Tax-1 and cellular proteins play a key role in HTLV-induced cell proliferation and genetic instability in vitro, and they facilitate viral persistence in vivo.  This provides new information regarding the distinct pathogenesis differences between HTLV-1 and HTLV-2. 

Unique Class of PCE Discovered in Retroviruses and junD

VO member Kathleen Boris-Lawrie, PhD, and her colleagues have discovered first in retroviruses, and subsequently in cellular proto-oncogene junD, a unique class of post-transcriptional control element (PCE).  Viral proteins are not required for PCE activity and host DEIH-box protein Dhx9/RNA helicase A (RHA), previously known to activate transcription, recognizes PCE structural features to facilitate polyribosome loading (Hartman et al, Nat Struc Mol Biol 2006;13(6):509-516). 

Oncolytic Viruses

Oncolytic viruses are either naturally emerging or engineered viral mutants with replicative selectivity towards tumor cells. These mutants include RNA and DNA, and it is likely that sequential combinations of different treatments will provide further patient life span extension. 
 
Despite various clinical trials, efficacy evidence has been elusive, human tumor tissue reveals that viral replication is minimal or nil, and a rapid host response consisting of infiltration and/or marker upregulation associated with mononuclear cells and lymphocytes occurs fairly rapidly.

VO investigators were among the first to observe a similar rapid response in animal models and to hypothesize that such a response was just one of several host responses whose role was to impede and limit the OV anticancer effect. The investigators have shown that several layers of host defenses must be circumvented for an OV successfully spread and kill neoplastic masses.

Collaborations

Viral Oncology  (VO) is one of OSUCCC–James' six highly interactive cancer Research Programs. The programs comprise more than 200 leading scientists from 13 different colleges within OSU and affiliated academic institutions.

VO investigators collaborate with several OSUCCC–James Shared Resources, including 
Analytical Cytometry, Biomedical Informatics, Biorepository & Biospecimen, Proteomics, Pharmacoanalytical, Nucleic Acid, Biostatistics and Microarray. The Program also maintains numerous collaborations with other OSUCCC Programs, with approximately 40 percent of more than 365 peer-reviewed publications having intra-programmatic and inter-programmatic collaborations. 

Other collaborations include:

Intra-Programmatic Collaborations

  • A collaborative study by VO Co-Leader Patrick Green, PhD, program member Michael Lairmore, DVM, PhD, and their research teams investigated the contribution of a novel nuclear b-ZIP protein (HBZ), which is encoded by the HTLV-1 genome antisense strand, to HTLV-1-mediated immortalization of primary T-lymphocytes in vitro and infection in a rabbit animal model. The study (Arnold et al, Blood 2006; 107:3976-3982) revealed that HBZ is not required for in vitro cellular immortalization, but it enhanced infectivity and persistence in inoculated rabbits. This investigation was the first to demonstrate that retroviruses utilize negative, strand-encoded proteins when chronic viral infections are established. 

Dr. Green and Dr. Lairmore also generated lentiviral vectors that express Hbz-specific short hairpin (sh)RNA to decrease Hbz mRNA and HBZ protein expression in transduced HTLV-1-transformed SLB-1 T-cells. Hbz knockdown correlated with a significant decrease in T-cell proliferation in culture, and both SLB-1 and SLB-1-Hbz knockdown cells engrafted into inoculated NOD/SCIDγchain-/- mice to form solid tumors that also infiltrated multiple tissues. Tumor formation and organ infiltration, however, were significantly decreased in animals challenged with SLB-1-Hbz knockdown cells.
Together these studies indicate that Hbz expression enhances the proliferative capacity of HTLV-1 infected T-cells, playing a critical role in cell survival and ultimately HTLV-1 tumorigenesis in the infected host (Arnold et al., Blood, 2008; 112(9):3788-97). 

  • VO Co-Leader Antonio Chiocca, MD, PhD, and VO members Robert Cavaliere, MD, and Balveen Kaur, PhD, have been pursuing development of an Adtk vector in Phase I and Phase II clinical trials as well as further development of this vector system to increase its immunogenicity and tumor vaccination effects.  Additionally, Experimental Therapeutics Research Program members Werner Tjarks, Ph.D., and Rolf Barth, MD are collaborating with VO members in an effort to add an additional tumor cell killing mode in this model.  By tk expression, they are working to target radiosensitizers for neutron-based radiation to brain tumors. 
  • VO member Balveen Kaur, PhD, in collaboration with VO Co-Leader Antonio Chiocca, MD, PhD, and Innate Immunity Research Program Co-Leader William Carson, MD,  secured a K01 award to study the combination of cilengitide with OV in brain tumors in a preclinical setting.
  •  VO Co-Leader Antonio Chiocca, MD, PhD, and program members Balveen Kaur, Ph.D., and Michael Caligiuri, MD, are collaborating to study the role of macrophages, microglia and NK cells in the tumor antiviral response. The researchers have shown that limiting each of these responses can increase OV therapy of malignant brain tumor. Additionally, a Dr. Chiocca received a U01 award from NINDS/NCI to perform preclinical toxicology studies of an oncolytic HSV1 in combination with the immunomodulating drug, cyclophosphamide (CPA), shown to reduce NK cell and macrophage/microglial activation and infiltration in an animal model of glioma, leading to improved therapeutic outcomes. 

Inter-Programmatic Collaborations

  • VO member Michael Lairmore, DVM, PhD, in collaboration with Molecular Carcinogenesis & Chemoprevention Research Program member Thomas Rosol, DVM, PhD, developed a bioluminescent mouse model and an effective therapy against adult T cell leukemia/lymphoma (ATLL) and humoral hypercalcemia of malignancy (HHM). The researchers utilized non-obese diabetic/severe combined immunodeficient (NOD/SCID) mice xenografted with ATLL cells to show that NFB inhibition by the proteasome inhibitor PS-341 and osteclast inhibition by zoledronic acid are effective ATLL and HHM treatments, which are refractory to conventional therapy (Shu et al, Cancer Research, 2007, 67(24):11859-66).  
  • In collaboration with Molecular Biology & Cancer Genetics Research Program member Daniel Schoenberg, PhD, VO member Kathleen Boris-Lawrie, PhD, collaborated with VO Co-Leader Patrick Green, PhD, and program member Michael Lairmore, DVM, PhD, and determined that a post-transcriptional control element (PCE) is mechanistically distinct from the internal ribosome entry sequence and promotes RNA translation that initiates translation by cap-dependent ribosome scanning. RHA is necessary for HTLV-1 gag RNA translation, and downregulation severely attenuates virion production (Bolinger et al., Nuc Acid Res 2007; 35(8):2629-42). The study results revealed a previously unidentified role for RHA in translation and implicated RHA as an integrated effector in the gene expression continuum from transcription to translation. Identifying PCE activity in eight human and animal retroviruses spanning six of seven Retroviridae genera and in cellular genes indicated that PCE is a conserved control mechanism for regulating translation of highly structured mRNA. The discovery is highlighted by the fact that two human cancer biomarkers are the upregulation of RHA and the accelerated translation in the tumor microenvironment.  Thus, RHA is an important molecule in human disease and is positioned as an attractive therapeutic target for lung, breast, ovarian and prostate neoplasms.   
  • The laboratories of Experimental Therapeutics Research Program member Said Sif, MSc, PhD, VO member Robert Baiocchi, MD, PhD, Innate Immunity Research Program Co-Leader John Byrd, MD, and Experimental Therapeutics Research Program Co- Leaders Samson Jacob, PhD, and Michael Grever, MD, have collaborated to clarify the consequences of protein arginine methyltransferase 5 (PRMT5) over expression on B cell transformation. The researchers discovered that the PRMT5 enzyme is over expressed in EBV-transformed lymphoblastoid cell lines and primary mantle cell lymphomas (EMBO J 2007 Aug 8;26(15):3558-69).  
  • VO member Robert Baiocchi, MD, PhD, Innate Immunity Research Program Co-Leader John Byrd, MD, and IIRP Program member Natarajan Muthusamy, DVM, PhD, are collaborating to clarify how PRMT5 becomes over expressed during EBV-driven B cell transformation.  Experiments with lentiviral expression systems designed to over express PRMT5 or a SiRNA that knocks down PRMT5 expression are addressing whether or not this protein is vital to the B cell transformation process.  
  • VO member Robert Baiocchi, MD, PhD, Experimental Therapeutics Research Program member Chenglong Li, PhD, and their teams are collaborating to utilize computational structural modeling to screen small molecules that will selectively target the PRMT5 enzyme.  To date, more than 50,000 compounds have been screened and eight promising small molecules are being evaluated in PRMT5 enzyme activity assays.  
  • VO members Robert Baiocchi, MD, PhD, and Pierluigi Porcu, MD, collaborated with Innate Immunity Research Program Co-Leader John Byrd, MD, IIRP member Michael Caligiuri, MD, and Experimental Therapeutics Research Program member Guido Marcucci, MD, and detailed how bortezomib induces DNA hypomethylation and silenced gene transcription by interfering with Sp1/NF-{kappa}B-dependent DNA methyltransferase activity in acute myeloid leukemia (Liu et al., Blood, 2008; 11(4):2364-73). The investigators demonstrated that bortezomib interferes with Sp1/NFkappaB binding and decreases Sp1 expression, resulting in reduced transactivation activity of Sp1/NFkappaB complex, reduced DNMT levels, DNA hypomethylation and silenced gene re-expression. These changes ultimately translate into antileukemic effects.

     Two Phase I trials with combinations of azanucleosides and bortezomib have been completed around this concepts. 
     
    Dr. Porcu, Dr. Caligiuri and Molecular Biology & Cancer Genetics Research Program member Gustavo Leone, PhD, also identified a specific tumor suppressor function for E2F2 in Myc-induced T cell lymphomagenesis (Opavsky et al., PNAS, 2007; 104(39):15400-5).

  • A collaborative investigation among VO members Ronald Glaser, PhD, Marshall Williams, PhD, Robert Baiocchi, MD, PhD, and Michael Caligiuri, MD, identified the Epstein-Barr Virus (EBV)-encoded deoxyuridine triphosphate nucleotidohydrolase (dUTPase) as an agent that leads to immune dysregulation. Recombinant and endogenous (EBV-LCLs) dUTPase leads to inhibition of peripheral blood mononuclear cells (PBMCs) proliferation/activation and the upregulation of several proinflammatory cytokines, including TNF-alpha, IL-1beta, IL-8, IL-6 and IL-10. CD14-positive cells (monocytes) depletion eliminated the cytokine profile induced by EBV dUTPase treatment. The data support the hypothesis that at least one protein of the EBV early antigen complex can induce immune dysregulation and may be involved in the pathophysiology of EBV-associated disease (Virology. 2006 346:205-18).   Recent characterization of this pathway by Dr. Glaser, Dr. Williams and their teams (J Immunol. 2009 182(2):851-9) demonstrated that dUTPase drives NF-kappaB activation in CD14+ macrophages via TLR2 and requires the recruitment of the adaptor molecule MyD88. 
  • EBV associated post-transplant lymphoproliferative disorder (EBV/PTLD) is directly related to a low frequency of EBV-specific cytotoxic T lymphocytes (CTLs) in patients who receive immunosuppressive therapy to prevent organ rejection. Collaborative efforts among VO members Michael Caligiuri, M.D., Robert Baiocchi, MD, PhD, and Pierluigi Porcu, MD, identified – via a chimeric mouse-human model of human PTLD (Baiocchi et al, J Clin Invest 108:887-894, 2001) and subsequently in PTLD patients (Porcu et al, Blood 100:2341-2348, 2002) – that PTLD tumors express abundant levels of the EBV lytic gene BZLF1 and that this gene product is an important target for adaptive cellular immune surveillance.

The researchers observed a correlation between BZLF1-specific CD3/CD8 T cell responses, tumor regression and overall survival in the preclinical model and in patients with PTLD.  They hypothesized that at least one component of the increased PTLD incidence in this patient population is the patient’s cellular immune deficiency against EBV lytic and latent antigens. A corollary to this hypothesis is that vaccine-enhanced EBV-specific immunity would restore the protection to this malignancy. They are currently developing a novel vaccination strategy against latent and lytic proteins expressed in EBV-associated PTLD using highly purified EBV polypeptides.

With support from private and non-profit organizations, larger in vivo trials are in progress with plans to move the first vaccine component through preclinical toxicity studies, to FDA for IND approval and then into a Phase I clinical trial. 

  • Over the past two years, VO members Robert Baiocchi, MD, PhD, and Pierluigi Porcu, MD, have assembled a multidisciplinary group of physicians and scientists to coordinate scientific and clinical work involving patients with EBV+ PTLD. This collaborative group, the PTLD Focus Group, includes physicians from the Department of Transplant Surgery and the Department of Internal Medicine (Divisions of Transplant Medicine, Nephrology, Infectious Disease, Hematology & Oncology). Currently, two protocols are being written to treat systemic PTLD and primary central nervous system PTLD.   
  • VO member Susan Wells, PhD, and her team study Human Papillomavirus oncogenic functions in viral replication and transformation. Of particular interest is the role and regulation of cellular transcriptional targets of the viral E6/E7 oncogenes. The human DEK gene is one induced E7 target, frequently upregulated in HPV-related and -unrelated human cancers. Results implicated DEK in cell death inhibition, cultured cell differentiation and 3D organotypic epithelial rafts (Wise-Draper, AJP, 2009).  Further, in a collaboration with fellow VO member Timothy Cripe, M.D., Ph.D., Dr. Wells defined DEK as a bona fide oncogene in classical transformation assays and in DEK knockout mice. Multiple members of the Fanconi anemia (FA) DNA repair pathway are also upregulated by E7. FA is a rare genomic instability syndrome characterized by congenital defects, progressive aplastic anemia and a predisposition to leukemia and squamous cell carcinoma. Thirteen FA genes have been identified to date, and the Wells laboratory demonstrated that the availability of multiple FA components can be co-regulated at the level of transcription and through Rb/E2F pathways (Hoskins, Oncogene, 2008). 
     Current collaborative experiments with Molecular Biology & Cancer Genetics Research Program member Gerard Nuovo, MD, and VO member Maura Gillison, MD, PhD, are designed to define FA pathway’s role in HPV DNA replication and tumorigenesis in FA patient-specific models of squamous cell carcinoma.

Inter-Institutional Collaborations

  • Cincinnati Children’s Hospital: VO members are collaborating with Cincinnati Children’s Hospital to foster development of a program project grant for oncolytic HSV therapy.  
  • Bone Marrow Transplantation Clinical Trials Network: VO member Robert Baiocchi, MD, PhD, and Steven Devine, M.D., are part of a national team of investigators from the BMT CTN who are developing autologous and allogeneic stem cell transplantation for patients with refractory/relapsed HIV-associated lymphomas. The first clinical protocol for autologous stem cell transplant is complete and being submitted to the IRB at EMMES Corp.   
  • AIDS & Cancer Specimen Resource: OSU is the Midwestern hub for the ACSR, an NIH-funded tissue and biological fluids bank of HIV-related specimens. Innate Immunity Research Program member Leona Ayers, M.D., is the principle investigator for the mid-region ACSR.   
  • University of Cincinnati and Cincinnati Children’s Hospital Medical Center: The HPV interest group among UC, CCHMC and OSU comprises multiple investigators focused on HPV transmission, epidemiology and vaccination, molecular biology and genomics, structural analyses, and clinical treatments.  
  • Eastern Cooperative Oncology Group and Radiation Therapy Oncology Group: Collaborations among VO member Maura Gillison, MD, PhD, her team, the ECOG and the RTOG have identified HPV infection as a strong risk factor for oral cancer and have determined that tumor HPV status is the single most important prognostic factor for head and neck cancer. Dr. Gillison has established the central laboratory at OSU for tumor HPV status testing for all cooperative group trials.  
  • National Cancer Institute USA and the Centers for Disease Control & Prevention:  In collaboration with the NCI and the CDC, VO member Maura Gillison, MD, PhD, and her team, along with the ECOG and the RTOG, have estimated the HPV-associated cancer burden and documented a dramatic increase in incidence rates for HPV-associated oral cancers in the United States. Dr. Gillison is the principal investigator of clinical trials specific to HPV-associated head and neck cancer, which will be performed collaboratively through the RTOG and EORTC at the direction of the NCI Task Force for Head and Neck Cancer.  
  • New Approaches to Brain Tumor Therapy Consortium: The VO has tested a genetically engineered mutant of adenovirus (designated ONYX-015) in a Phase I, dose-escalating clinical trial as part of NABTT, a multi-institutional, NCI-funded consortium.

The Ohio State University Comprehensive Cancer Center – Arthur G. James Cancer Hospital and Richard J. Solove Research Institute (OSUCCC – James) 460 W. 10th Avenue, Columbus, OH 43210 Phone: 1-800-293-5066 | Email: jamesline@osumc.edu