Laboratory of Papillomavirus Research
Investigator, Center for Oncology and Cell Biology
Professor and Dean, Elmezzi Graduate School of Molecular Medicine
Chief of Otolaryngology Research, Long Island Jewish Medical Center

350 Community Drive
Manhasset, NY 11030
Tel: 516-562-1159
Fax: 516-562-1022
Email: bsteinbe@lij.edu

Academic Degrees
B.A. 1959, University of California, Riverside
M.S. 1967, Adelphi University
Ph.D. 1976, State University of New York at Stony Brook

Recurrent respiratory papillomatosis (RRP) is a rare but very serious disease caused by a human papillomaviruses (HPVS), a family of viruses that cause a wide range of benign and malignant tumors ranging from common skin warts and genital warts to cervical cancer and some cancers in the oral cavity.  Respiratory papillomas are generally benign, but can become malignant.  The only approved treatment for recurrent papilloma growth is repeated surgical removal to prevent them from blocking the airway – for some patients it’s a monthly trip to the operating room.  Improved therapies, based on better knowledge of the biology of these HPVs, are badly needed.  The Steinberg laboratory, in collaboration with Dr. Marc Symons of the Feinstein Institute, is studying HPV-target cell interactions, with particular focus on altered signal transduction that results in the expression of the enzyme COX-2 and its role in activation of latent HPV infection.  This enzyme is also expressed in many cancers, and may promote tumor growth.  We are also conducting a clinical trial of celecoxib, a COX-2 inhibitor, to determine whether if will be an effective therapy for RRP, in collaboration with Dr. Allan Abramson  and Dr. Mark Shikowitz, chairman and vice chairman respectively, of the Department of Otolaryngology at Long Island Jewish Medical Center.  We are also collaborating with Dr. Vincent Bonagura  of the Feinstein Institute on studies of altered host immune responses to HPV, resulting in recurrent disease.  Finally, we are collaborating with Dr. Samuel Soffer of the Feinstein Institute on ways to inhibit blood vessel growth in tumors.

Research Projects:

Recurrent respiratory papillomatosis (RRP) is a mucosal disease of the airway caused primarily by human papillomaviruses (HPVs) -6 and -11.  RRP is characterized by growth of multiple papillomas that recur following surgical removal.  Disease can last for the life of the patient or can be interspersed by short or long periods of remission.  Recurrence rates vary between patients, with the worst cases requiring surgery as frequently as every three weeks to maintain an airway.  Fortunately, the prevalence of the disease is low, estimated to be approximately 3/100,000 in the United States.  The papillomas are primarily located in the larynx, but approximately 17 percent of patients will have tracheal disease and 5 percent will have papillomas of the bronchus and lung parenchyma.

Respiratory papillomas are usually benign, with eventual malignant conversion in 3% to 4% of patients, but anecdotal evidence suggests that the conversion rate in pulmonary papillomas may be as high as 80%.  We are conducting a number of studies to better understand RRP, and to develop better therapies for the disease:

Defects in control of growth and differentiation.  We have found that papilloma cells are characterized by a proliferative rate that is no higher than uninfected cells.  Rather, they have a defect in terminal differentiation that results in an accumulation of cells that have left the cell cycle but have minimal expression of normal differentiation-specific keratins and filaggrin. The cells are also relatively resistant to apoptosis.  Studies are continuing to determine the molecular mechanism(s) of these phenotypes.

Altered Signal Transduction and Expression of COX-2.  We have found that papilloma cells and respiratory papilloma tissues express high levels of the EGF receptor (EGFR), and constitutive activation of the EGFR causes the defect in differentiation.  Papilloma cells also have alterations in several signaling pathway intermediates linked to the EGFR.  These include activation of PI 3-kinase, overexpression of Rac1, activation of Pak1 and Pak2, activation of p50 NF-κB, and activation of p38 MAP kinase.  These studies are continuing, in collaboration with Dr. Marc Symons and his laboratory.  Activation of the signal transduction intermediates result in constitutive expression of the enzyme COX-2 and its product PGE2.  Inhibition of COX-2 or some of the upstream intermediates in papilloma cells causes slowed proliferation and increases spontaneous apoptosis.  We are currently asking whether COX-2 and PGE2 contribute to the defect in differentiation, and whether PGE2 contributes to constitutive activation of the signaling intermediates through a positive feed-back loop.

Clinical trial of celecoxib for treatment of RRP.  Celecoxib is a selective COX-2 inhibitor.  Based on the finding that celecoxib inhibited papilloma cell growth and enhanced apoptosis, and a literature showing that these potent anti-inflammatory drugs were effective in certain cancers, we conducted a preliminary clinical treatment study on three patients with severe RRP. The results were striking. The papillomas stopped recurring during the one-year of treatment, and improvement was maintained after the treatment was discontinued. We are now conducting a multi-centered double-blind clinical trial testing the benefits of celecoxib as an adjunct to surgery. The study is being conducted in collaboration with Dr. Allan Abramson and Dr. Mark Shikowitz at Long Island Jewish Medical Center, and physicians at Eastern Virginia Medical Center, Norfolk; University of Iowa; University of Alabama, Birmingham; University of California, San Francisco; and Sanford Medical Center, Sioux Falls, ND.

Persistence of latent HPV infection.  Patients with RRP (or patients with a history of RRP that are now in remission) have widespread latent HPV infection in clinically normal tissues of the larynx, trachea and bronchi, with life-long persistence of viral DNA but essentially no expression of viral RNA and no clinical or histologic evidence of disease.  We propose that recurrent papillomas are due to repeated activation of this latent infection, rather than re-infection or “seeding” of the virus.  The mechanism of activation is not known, but trauma or irritation facilitate or enhance activation in model systems.  We are currently asking whether induction of COX-2 during transient inflammation or irritation mediates activation of latent HPV DNA. 

Altered host immune response to HPV.  We are collaborating with Dr. Vincent Bonagura and his laboratory in their studies to determine why patients with RRP don’t generate an effective immune response to prevent recurrent disease.  We are trying to understand how the ubiquitous human papillomavirus can remain latent, or dormant, in most people but cause respiratory papillomas in a few.  This question has broad implications beyond respiratory papillomatosis, since latent HPV infections are present in the airway, skin and genital tract of many people and may be a source of subsequent skin warts, genital warts, and cervical and airway cancers.  We have found that patients with RRP have a bias toward a TH2-like response to HPV proteins, have elevated levels of Treg cells in the papillomas, and show altered expression of a number of innate and adaptive immune response genes.  These altered responses may be, at least in part, genetic.  Patients have a skewed distribution of HLA Class II alleles, and may lack some activating Kir gene alleles.  They may also express higher levels of COX-2 in their airway tissues, which could bias the local immune response toward expression of TH2-like cytokines and chemokines, reduce clearance of active infection, and make the patients more likely to have the disease.  

Tumor angiogenesis and inhibition.  Papilloma tissues are highly vascularized, and express VEGF.  In this respect, they are similar to many cancers.  The VEGF may be induced by PGE2 expressed in the papillomas.  We are collaborating with Dr. Sam Soffer in his studies of angiogenesis and the molecular basis for the use of drugs to inhibit angiogenesis. Current studies focus on use of rapamycin in a sarcoma model.  This drug, at low doses, suppresses tumor growth.  It may act by inducing apoptosis of vascular endothelial cells in the tumor and possibly by inhibiting expression of COX-2 by the sarcoma cells.

Lab Members:

Name: Rong Wu, MD, PhD
Position:   Assistant Investigator, Center for Oncology and Cell Biology
Education:  M.D., 1987, China Medical University, Shengyang, China
Ph.D., 2000, Hamamatsu University School of Medicine, Hamamatsu, Japan
Research:   Studies alterations in signal transduction pathways in recurrent respiratory papillomas, and the positive COX-2 feedback loop.  Currently investigating the activation of the EGFR, PI3K and Rac1 by PGE2.
E-mail:   rwu1@nshs.edu

Name:   Aaron Lipskar, MD
Position:   Research Fellow
Education:   B.A., 1998,  Brandeis University M.D., 2004, Temple University School of Medicine
Research:   Studies signal transduction  and angiogenesis associated with mTOR, a target of rapamycin, a drug used for treating tumors.
E-mail:   alipskar@nshs.edu

Name:   Ray Pica
Position:   Senior Research Assistant
Research:   Manages the laboratory, conducts experiments and supervises volunteers. Currently making and analyzing a number of HPV11 mutant constructs to determine which viral proteins induce COX-2 in epithelial cells.
E-mail:   rpica@nshs.edu

Name:   Michael Hall
Position:  Graduate  Student
Education:   B.A., 2007,  M.S., 2009, Boston University (anticipated)
Research:   Studies effects of COX-2 and PGE2 on epithelial differentiation in normal and HPV-infected cells.
E-mail:  mh222@bu.edu

Name:   Tomoki Nomakuchi
Position:   Student volunteer
Education:  B.S., 2010, Hofstra University (anticipated)
Research: Studying altered differentiation in HPV-infected cells
E-mail: tnomak1@pride.hofstra.edu

Selected Publications:

Devoti, J.A., Rosenthal, D.W., Wu, R., Abramson, A.L., Steinberg, B.M., Bonagura, V.R.  Immune Dysregulation and Tumor-Associated Gene Changes in Recurrent Respiratory Papillomatosis: A Paired Microarray Analysis. Mol Med. 14: 608-617, 2008.

Wu, R., Coniglio, S.J., Chan, A., Symons, M.H., Steinberg, B.M. Up-regulation of Rac1 by Epidermal Growth Factor Mediates COX-2 Expression in Recurrent Respiratory Papillomas. Mol Med 13:143-50, 2007.

Wu, R., Abramson, A.L., Shikowitz, M.J., Dannenberg, A.J., and Steinberg, B.M.  Epidermal Growth Factor-Induced Cyclooxygenase-2 Expression is Mediated through Phosphatidylinositol-3 Kinase, not Mitogen-Activated Protein/Extracellular Signal-Regulated Kinase Kinase, in Recurrent Respiratory Papillomas.  Clinical Cancer Research, 11: 6155-6161, 2005.

Dackour, R., Carter, T., Steinberg, B.M. Phosphatidylinositol 3-Kinase Regulates Early Differentiation In Human Laryngeal Keratinocytes.  In Vitro, Cell Dev. Biol.  41:111-117, 2005.

Bonagura, V.R., Vambutas, A., DeVoti, J.A., Rosenthal, D.W., Steinberg, B.M., Abramson, A.L., Shikowitz, M.J., Gjertson, D.W., Reed, E.F. HLA Alleles, IFN-γ Responses to HPV-11 E6, and Disease Severity in Patients with Recurrent Respiratory Papillomatosis.  Human Immunol 65:773-82, 2004.

DeVoti, J.A., Steinberg, B.M., Hatam L, Vambutas A, Abramson A.L., Shikowitz, M.J., Rosenthal, D., Bonagura, V.R.  Failure of IFN-γ, But Not IL-10 Expression, In Response to HPV-11 E6 Protein In Respiratory Papillomatosis.  Clinical and Diag. Lab. Immunol. 11:538-47, 2004.

Vambutas, A., Bonagura, V.R, Reed, E.F., Abramson, A.L., Mullooly, V., DeVoti, J., Gjertson, D.W., Steinberg, B.M.  Polymorphism of transporter associated with antigen presentation 1 as a potential determinant for severity of disease in recurrent respiratory papillomatosis caused by human papillomavirus types 6 and 11. J Infect Dis.189:871-9, 2004.

Abramson, A.L., Nouri, M., Mullooly V. and Steinberg, B.M. Latent Human Papillomavirus infection is comparable in the larynx and trachea. J. Med. Virol. 72:473-477, 2004.

Wu, R., Sun, S., Steinberg, B.M.  Requirement of STAT3 activation for differentiation of mucosal stratified squamous epithelium.  Mol Med.9:77-84, 2003.

Sun, S. and Steinberg, B.M. Identification of PTEN as a negative regulator of STAT3 activation in human papillomavirus-infected cells. J. Gen Virol. 83:1651-58, 2002.

Vambutas, A., DeVoti, J., Pinn, W., Steinberg, B.M. and Bonagura, V.R. Interaction of Human Papillomavirus Type 11 E7 Protein with TAP-1 results in reduction of ATP dependent peptide transport.  Clinical Immunol. 101:94-99, 2001.

Vambutas, A., Bonagura, V.R. and Steinberg, B.M. Altered expression of TAP-1 and Major Histocompatibility Complex Class I in laryngeal papillomatosis.  Correlation of TAP-1 with disease. Clin. and Diag. Lab. Immunol. 7:79-85, 2000

Zhang, P. and Steinberg, B.M. Overexpression of PTEN/MMAC1 and decreased activation of Akt in HPV-infected laryngeal papillomas.  Cancer Res. 60:1457-1462, 2000.

Zhang, P., Nouri, M., Brandsma, J.L., Iftner, T. and Steinberg, B.M. Induction of E6/E7 expression in cottontail rabbit papillomavirus latency following UV activation.  Virology 263:338-394, 1999.

Bonagura, V.R., Hatam, L., DeVoti, J., Zeng, F. and Steinberg, B.M. Recurrent respiratory papillomatosis: Altered CD8+ T-Cell Subsets and TH1/TH2 cytokine imbalance.  Clinical Immunol. 93:302-311, 1999.

Johnston, D., Hall, H., DiLorenzo, T.P. and Steinberg, B.M. Elevation of the Epidermal Growth Factor Receptor and Dependent Signaling in Human Papillomavirus Infected Laryngeal Papillomas.  Cancer Res. 59:968-974, 1999.

Maran, A., Amella, C.A., DiLorenzo, T.P., Auborn, K.J., Taichman, L.B. and Steinberg, B.M.  Human Papillomavirus Type 11 Transcripts Are Present at Low Abundance in Latently Infected Respiratory Tissues.  Virology 212:285-294, 1995.

Bonagura V.R., Siegal F.P., Abramson A.L., Santiago-Schwarz F., O'Reilly M.E., Shah K., Drake D. and Steinberg B.M.  Enriched HLA-DQ3 Phenotype and Decreased Class I Major Histocompatibility Complex Antigen Expression in Recurrent Respiratory Papillomatosis.  Clin. & Diag. Lab. Immunol. 1:357- 360, 1994.

Amella C.A., Lofgren L.A., Ronn A.M., Nouri M., Shikowitz M.J. and Steinberg B.M.  Latent Infection Induced with Cottontail Rabbit Papillomavirus: A Model for Human Papillomavirus Latency.  Am. J. Pathol. 144:1167-171, 1994.

Vambutas, A., DiLorenzo, T.P. and Steinberg, B.M.  Laryngeal Papilloma Cells Have High Levels of Epidermal Growth Factor Receptor and Respond to Epidermal Growth Factor by a Decrease in Epithelial Differentiation.  Cancer Research 53:910-914, 1993.