Treatment of HPV positive penile and cervical cancer by induction of HPV...


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Title Treatment of HPV positive penile and cervical cancer by induction of HPV E7-specific T cell immunity through DNA vaccination
Period 06 / 2006 - unknown
Status Completed
Research number OND1318072
Data Supplier ZonMw


Human papilloma virus infection (serotypes 16 and 18) is strongly associated with the development of squamous cell cancer, such as head-and neck cancer, cervical cancer, vulvar and penile carcinoma, and anal carcinoma. Because the persistence of oncogenic HPV proteins E6 and E7 is required for carcinogenesis, these viral antigens are exquisite targets for immunotherapeutic interventions. Indeed, therapeutic vaccinations targeting these viral antigens have shown some promise in woman suffering from cervical cancer. Here, we propose to initiate a phase I/II study in patients with HPV 16-positive squamous cell cancer of the penis and cervix using a novel and potent intradermal DNA vaccination strategy. In preclinical studies this strategy was shown to be much more potent in the induction of E7-specific T cell immunity than existing non-viral vaccination strategies, providing a strong rationale for its clinical evaluation. In the phase I part of the clinical trial, the toxicity of this novel DNA vaccination strategy will be evaluated in advanced-stage penile cancer patients. If no major toxicities are found, the phase II part of the study will define the immunological efficacy of this highly promising DNA vaccination strategy in both high risk penile cancer patients and recurrent cervical cancer patients. This will allow us to define the value of this novel DNA vaccination strategy for the treatment of HPV-associated malignancies. Persistent Human Papilloma Virus (HPV) infections (largely serotypes 16 and 18) are associated with development of squamous cell cancer such as head-and-neck cancer (Braakman et al., 2004) cervical cancer, vulvar, anal (Carter et al., 2001) and penile cancer (Ferreux et al., 2003). HPV-positive cancers express the viral oncogenic proteins E6 and E7. Sustained expression of these proteins is required to preserve the malignant state. HPV protein E7 interacts with the cell cycle regulator retinoblastoma protein, pRb, thereby allowing and promoting entry in the S-phase of the cell cycle. HPV E6 destabilizes p53, which is a key protein in cell cycle control, thereby preventing cell cycle arrest and apoptosis. Because continued expression of HPV E6 and E7 is required for carcinogenesis and because of their viral origin, these proteins are attractive targets for immunotherapeutic interventions. In fact, for cervical cancer several therapeutic vaccination trials are ongoing, targeting HPV 16/18 E6 and/or E7 proteins. So far, such studies have not yet been initiated for other HPV associated cancers. While E6 and E7 are viral gene products and therefore are not subject to immunological tolerance mechanisms, powerful vaccination strategies are nevertheless required to mount the sizeable cellular immune responses that are necessary to target pre-existing tumor lesions. In the past years we have developed a novel vaccination strategy using a multiple micro needle injection technique for intradermal plasmid DNA application, called DNA tattooing. This strategy was optimized based on our analysis of in vivo antigen expression kinetics and ex vivo immunomonitoring. In preclinical models, DNA tattooing has proven to be a rapid and robust strategy to induce vaccine-specific cellular and humoral immune responses. Importantly, in a direct comparison, DNA tattooing is by far superior to other non-viral means of vaccination (Bins et al., 2005 in press). Safety studies with DNA tattooing in large animal models are now ongoing and upon completion of these studies we intend to define its potency in a phase I/II clinical trial with HPV 16-associated penile and cervical cancer. Since the NKI-AVL is a centre hospital for penile cancer in the Netherlands and the LUMC for cervical cancer, both penile and cervical cancer are excellent candidate diseases for clinical testing of HPV E7 DNA tattooing. Encouraging results with regard to safety and efficacy will be followed up by the exploration of multineedle DNA vaccination in other HPV-associated neoplasia, such as vulvar intra-epithelial neoplasia (VIN), and will include an efficacy comparison with peptide-based vaccines that are currently in phase I trials in cervical cancer patients. This project proposal aims at translating a novel DNA vaccine-based strategy to clinical application in patients with HPV 16-positive penile and cervical squamous cell carcinoma. In the pharmaceutical phase the recently built GMP production facility will be validated for production of clinical grade DNA vaccines. Note: Production of the required material for validation of production facility is ongoing. This part of the research plan is therefore likely to be completed prior to initiation of this project. In the phase I part of the study we aim to determine the safety of this DNA vaccination strategy in patients with HPV 16-positive penile cancer. Having established the safety profile of the strategy, the multicenter phase II part of the study will be opened to establish the efficacy of this strategy in inducing HPV-specific cellular immune responses in patients with high risk penile cancer and recurrent cervical cancer. The immunomonitoring will be performed in a GLP laboratory dedicated to measure HPV-specific cellular immune responses. We have developed a novel DNA vaccination strategy in a mouse model that results in very robust vaccine-specific immune responses (Bins et al., 2005 in press). The strategy was designed based on the measurement of in vivo antigen-expression using a light-emitting reporter gene. A short-interval intradermal DNA vaccination by multiple micro-needle injections, called DNA tattooing, was shown to be extremely efficient in inducing strong vaccine-specific T cell responses in a short time frame. Skin due to its physiologic barrier function is likely to be a preferred site for DNA vaccine delivery compared to muscle due to the natural presence of immunological defence mechanisms. In addition, translation of vaccination strategies from mouse to man is likely to be more feasible for a 2D (surface-based) intradermal vaccination instead of the volume-based intramuscular DNA vaccination. To examine the potency of DNA tattooing in humans, a phase I/II study will be performed in patients with advanced-stage and high-risk HPV-positive penile and recurrent cervical squamous cell carcinoma. DNA vaccines with a non-oncogenic E7 gene will be produced under GMP and will be employed in this study. The current project will use an HPV 16 E7 encoding DNA vaccine to induce E7-specific T cell immunity in advanced-stage HPV16+ penile and cervical cancer patients. The clinical phase I study will be performed in advanced-stage penile cancer patients. Once the safety of the strategy has been demonstrated, the phase II part of the study will be initiated. The number of penile cancer patients that are yearly referred to the NKI-AVL for treatment and that qualify for participation in this trial may be not large enough to allow rapid accruel of patients. Therefore, we have decided to extend the inclusion criteria to allow enrollment of cervical cancer patients as well. We have chosen to target the E7 protein of the HPV 16 subtype for several reasons. Firstly, HPV 16 is the most common virus associated with both cervical and penile cancer. Secondly, E7 (as well as E6) is consistently expressed in HPV induced carcinomas. Thirdly, both CD4+ and CD8+ E7-specific T cell epitopes are present within the E7 oncoprotein (for example: HLA-A*0201 binding epitopes E7(11-20), E7(82-90), E7(86-93) (Kast et al., 1994)), making the HPV 16 derived E7 protein a good candidate vaccine.

Related organisations

Secretariat The Netherlands Cancer Institute
Financier ZonMw

Related people

Researcher Dr. B. Nuijen
Researcher Prof.dr. T.N.M. Schumacher
Project leader Prof.dr. S.H. van der Burg
Project leader Prof.dr. J.B.A.G. Haanen
Project leader Prof.dr. S. Horenblas
Project leader Prof.dr. C.J.M. Melief


D21800 Immunology, serology
D23120 Oncology

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