Tina Dalianis' Group
Studies on human papillomaviruses and specific markers for tailored and targeted therapy of head and neck cancer.
Background and previous results
There are >150 HPV types of which some types or species are potentially oncogenic. Best known are HPV16 and 18, which cause cervical cancer and included in today's HPV vaccines. However, besides smoking and alcohol, HPV16 has a causal link to oropharyngeal cancer, where tonsillar and base of tongue cancer (TSCC and BOTSCC) dominate. We and others have shown that HPV-positive TSCC and BOTSCC have a much better survival than corresponding HPV-negative cancers and head/neck cancers in general (80% vs. 40% 5-year disease-specific survival). In addition, we were the first to show that the incidences of HPV-positive TSCC and BOTSCC have increased epidemically the past decades. Due to its poor prognosis, therapy for head and neck squamous cell carcinoma (HNSCC) generally includes chemotherapy and intensified radiotherapy (RT), with many more side effects compared to conventional RT. This is unnecessary for 80% of patients with HPV-positive TSCC and BOTSCC who do well with conventional RT and account for >25% of all HNSCC. However, before de-escalating therapy, it is important to find markers that along with HPV can better predict a favorable response to conventional treatment.
Preliminary results and research plan
In a TSCC and BOTSCC patient cohort from 2000-2011 at the Karolinska University Hospital, we have identified such markers (e.g. HLA class I, CD44, LRIG, CD8 tumour-infiltrating lymphocyte counts, expression of HPV16 E2 mRNA), which individually or together identify 20-30% of patients with 90-95% probability for complete response. Using two mathematical models we have combined HPV-positive status with several prognostic biomarkers and can identify 40-56% of the patients that respond to therapy. Now we want to find additional markers to be able to combine old and newer markers in our mathematical models and understand their mechanisms of action to identify virtually all patients with a very good outcome. Moreover, during our studies we have disclosed that intensified therapy does not improve survival for those that do not respond to therapy and better alternatives e.g. targeted therapy are needed. Our research will will enable us to identify patients that respond to therapy safely and to start prospective randomized trials with less therapy. For those that do not respond to therapy, we hope to find other alternatives such as specific targeted therapy.
One step in this direction and to find additional biomarkers was to examine both HPV-positive and HPV-negative tumors for mutations in oncogenes and suppressor genes and to perform protein profiling. A second step just initiated with Dr Juan Du and prof. Lars Engstrand is to test our tumours for specific microbiome profiles, should they be useful as biomarkers.
Examining hotspot mutations by Next Generation Sequencing of 50 cancer related genes in 300 HPV+ TSCC/BOTSCC and 50 HPV- TSCC/BOTSCC and found that the genes PIK3CA and FGFR3, both targetable proteins, were among the most mutated ones in HPV+ TSCC/BOTSCC, while p53 was most frequently mutated in HPV-TSCC/BOTSCC (Fig.1). Notably, we also found that mutated FGFR3 in HPV positive tumours correlated to worse prognosis.
This indicates that FGFR3 could be very useful as an additional predictive marker to combine with the markers already mentioned earlier in the mathematical models described above to distinguish patients with very favourable or poor prognosis. Furthermore, 36% of those with FGFR3 mutations had PIK3CA mutations, thus possibly targetable with two drugs.
When performing protein profiling done by a sensitive proximity elongation assay (PEA) we found that some proteins related to angiogenesis, e.g. VEGF-A, CYR61 and ESM-1 and IGF1R, the latter conferring resistance to EGFR inhibitors, are up-regulated, while hK-8 is low, in HPV+ TSCC/BOTSCC that do not respond to treatment, indicating that these proteins can be used to predict effect of therapy.
We will now include the newly disclosed biomarkers in our previous cohort from 2000-2011 in the different mathematical models to examine if additional patients with a very good prognosis can be identified. Moreover, we are presently collecting a new TSCC and BOTSCC cohort from 2012-2016 from the Karolinska University Hospital, to validate all new markers (FGFR3, PIK3CA, VEGF-A etc.) by sequencing and/or protein profiling, and to examine if can be detected by immunohistochemistry.
In parallel, we are setting up in vitro models tissue culture models for targeted therapy. HPV positive and HPV negative TSCC and BOTSCC cell lines, we have and will obtain (around 5 of each) will be tested for presence of the above mutations or protein deregulation patterns. If the specific mutations of interest e.g. FGFR3 and PIK3CA are not already in our cell lines, they will be introduced with CRISP/Cas9 technology. This way we hope to identify whether drugs used today for targeted therapy in other types of tumours also can be used for TSCC and BOTSCC. Useful drugs will also be further analyzed in experimental animal PDX systems.
By delivering novel tools to better tailor therapy for growing numbers of patients with TSCC and BOTSCC, allowing some patients to receive tapered therapy, and others to receive more specific targeted therapy we will significantly increase survival as well as quality of life, issues of both ethical and socioeconomic importance.
Nathalie Hou Grün
"Human papillomavirus infection in healthy youth and in hypopharyngeal cancer"
June 3, 2016
"HLA-A*02 and its prognostic traits in cancer : an immunological biomarker as a tool towards individualised cancer therapy"
February 5, 2016
"Hunting the end of the rainbow : prognostic biomarkers and human papillomavirus in tonsillar and base of tongue cancer"
January 15, 2016
"Studies on human papillomavirus and molecular markers in head-neck cancer"
April 24, 2015
"Studies of the influence of human papillomaviruses (HPV) and other biomarkers on the prevalence of oropharyngeal cancer and clinical outcome"
May 31, 2013
"Studies on human papillomaviruses in head and neck cancer"
February 1, 2013
"Occurrence of human papillomaviruses (HPV) types in HPV related cancer and in the genital and oral tracts of young adults"
September 14, 2012
"Cancer therapy using viral- and bacterial proteins, as vectors for vaccines or as carriers of cytostatics"
March 30, 2012
"Studies on polyomaviruses in humans : In relation to haematopoietic stem cell transplantation and cancer"
February 26, 2010
"Vaccination against Her2/neu - expressing cancer using chimeric virus-like particles"
May 15, 2009
"Studies on the occurence and effects of human papillomavirus in tumors of the head and neck"
March 28, 2008
"Studies on polyomavirus virus-like particles : As vaccines and vectors for immune and gene therapy"
March 31, 2006
"Studies on the presence and influence of human papillomavirus (HPV) in head and neck tumors"
April 8, 2005
Andrea Vlastos Franzén
"Murine polyomavirus VP1 virus-like particles as vectors for gene therapy and as vaccines against polyomavirus infection and tumors"
June 11, 2004
Hanna Mellin (Dahlstrand)
"Human papillomavirus in tonsillar cancer"
November 22, 2002
"Studies on human polyomavirus infection in immunosuppressed patients with polyoma related tumors"
October 12, 2001
"Studies on persistant polyomavirus infection in relation to tumor development and options for vaccine and gene therapy"
June 1, 2001
Tina Dalianis, Professor, Group leader
Torbjörn Ramqvist, Assistant Professor
Cinzia Bersani, PhD, Post doc
Andrea Vlastos, MD, PhD, ENT specialist
Anders Näsman, MD, PhD, specialist in Clinical Pathology
Lars Sivars, MD, PhD student
Linnea Haeggblom, PhD student
Andreas Ährlund-Richter, student
Joar Franzén, student
Juan Du, PhD, Associated
Group photo 2014
Group photo 2012
Group photo 2006
Activities at KI Summer Research School 2005