Studies on human papillomaviruses and specific markers for tailored and targeted therapy of head and neck cancer and childhood cancer – Tina Dalianis' Group

Human papillomavirus positive oropharyngeal cancer is increasing in incidence and despite its good prognosis not all patients are cured. Likewise, for high risk neuroblastoma and medulloblastoma not all cancers are cured. We focus on finding biomarkers that are prognostic and predictive for treatment responses e.g. to immune check inhibitors and that are also useful for targeted therapy. We have promising data on synergistic effects with targeted therapies against the above cancers.

Studies on 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 severe side effects. Intensified therapy has not improved survival for those with a poor prognosis and some patients are likely overtreated. Clearly therefore, new therapeutic options are needed for this group of patients that today account for >40% of all HNSCC.  However, to better individualize therapy it is important to find markers that along with HPV can better predict a favorable response to conventional treatment, or that can be useful for targeted therapy. 

Previous data and research plan

In a TSCC and BOTSCC patient cohort from 2000-2011 at the Karolinska University Hospital, we identified most but not all such markers using immunohistochemistry (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 mathematical models we later combined HPV-positive status with several prognostic biomarkers and can identify 40-56% of the patients that respond to therapy.

We then proceeded to find additional markers using DNA and RNA sequencing, some data are described below.

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. Furthermore, others have also reported that when de-escalating treatment patients with tumours with PIK3CA mutations did worse.

The above data indicated that targeting PI3K and FGFR3 could potentially be useful for this patient group.

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.

More recently, we performed whole exome sequencing (WES) on tumors of patients with and without relapse and found some genes, e.g. a specific CDC27 deletion variant that was present only in patients that had encountered a relapse, while others such as BLAF1 and AQP1 as well as CDC27 had mutations in their genes both cohorts. Hopefully some of the data here will contribute to finding new options for targeted therapy.

Fig. 1 A. Variants on the y axis. Red denoting patients with recurrence; blue their metastasis; green patients without recurrence.
Fig. 1 B. Genes mutated in >30% of the whole cohort of patients on the y-axis. Red: patients with recurrence; green patients without recurrence.

To acquire more data on targeted therapy, we have employed in vitro models tissue culture models for targeted therapy. HPV positive and HPV negative TSCC and BOTSCC cell lines with or without e.g. FGFR3 and PIK3CA mutations have been obtained. These TSCC/BOTSCC cell lines and have been tested for their sensitivity to different targeted therapies using inhibitors against PI3K, FGFR, CDK4/6, PARP and WEE1 inhibitors by viability, proliferation, apoptosis, cytotoxicity and FACs assays.

These experiments that have been performed were very promising. We have for example found that our cell lines show dose dependent responses to most of the inhibitors. However, when some of these  inhibitors are combined, both additive and synergistic effects were disclosed. We are now combining these inhibitors with irradiation or chemotherapy and studying their joint effects.

With this approach we hope to identify whether drugs used today (indicated above) for targeted therapy in other types of tumors, or against newly disclosed mutated genes by WES also can be used for TSCC and BOTSCC. Useful drugs will also be further analyzed in experimental animal PDX systems.

Moreover, we are presently barcoding our cell lines in order to follow subclonal evolution after various drug treatment and performing single DNA and RNA sequencing to study resistant patterns according to the specific molecular profile of each tumour cell line.

Finally, we have just initiated some additional studies on HPV viral load and cell free HPV DNA in order to follow prognosis and possible risk for relapse.

Studies of childhood cancer

In parallel with our studies above, we have also screened a number of childhood cancers such as a number of different childhood brain tumors and neuroblastomas for the presence of FGFR3 and PIK3CA mutations. These types of mutations are rare, but have been found in at least one neuroblastoma (NB). We are therefore also testing some NB cell lines with the same assays that have been described above for the tonsillar and base of tongue cancer (TSCC and BOTSCC) cell lines. We have found some positive effects and these are investigated further by team Ourania Kostopoulou in our group.


Using novel tools to better prognosticate and tailor therapy for growing numbers of TSCC/BOTSCC patients, it will be possible offer targeted therapy for those with poor prognosis and avoid overtreatment of those with good prognosis, which will thereby increase survival as well as quality of life.  

For childhood cancer, if targeted therapy can be used synergistically this may be useful for those with tumors that are resistant to conventional chemotherapy and help increase survival as well as avoid some side effects associated with conventional chemotherapy.

PubMed publications from Tina Dalianis

Staff and contact

Group leader

All members of the group


Mark Zupancic
"Diagnostic and prognostic markers in primarily non-smoking related head and neck cancer"
June 16, 2023

Andreas Ährlund-Richter
"Studies on human papillomavirus (HPV) and other markers in the development and prognosis of HPV associated cancer"
June 14, 2022

Linnea Haeggblom
"Human papillomavirus and other molecular markers in cancer of different oropharyngeal sub-sites"
December 7, 2018

Lars Sivars
"Human papillomavirus and other biomarkers in neck masses of unknown origin and in head and neck cancer"
May 9, 2018

Nathalie Hou Grün
"Human papillomavirus infection in healthy youth and in hypopharyngeal cancer"
June 3, 2016

Lisa Villabona
"HLA-A*02 and its prognostic traits in cancer : an immunological biomarker as a tool towards individualised cancer therapy"
February 5, 2016

Nikolaos Tertipis
"Hunting the end of the rainbow : prognostic biomarkers and human papillomavirus in tonsillar and base of tongue cancer"
January 15, 2016

Michael Mints
"Methodological approaches towards personalised cancer medicine"
October 23, 2015

Cecilia Nordfors
"Studies on human papillomavirus and molecular markers in head-neck cancer"
April 24, 2015

Anders Näsman
"Studies of the influence of human papillomaviruses (HPV) and other biomarkers on the prevalence of oropharyngeal cancer and clinical outcome"
May 31, 2013

Mircea Romanitan
"Studies on human papillomaviruses in head and neck cancer"
February 1, 2013

Juan Du
"Occurrence of human papillomaviruses (HPV) types in HPV related cancer and in the genital and oral tracts of young adults"
September 14, 2012

Mathilda Eriksson
"Cancer therapy using viral- and bacterial proteins, as vectors for vaccines or as carriers of cytostatics"
March 30, 2012

Géraldine Giraud
"Studies on polyomaviruses in humans : In relation to haematopoietic stem cell transplantation and cancer"
February 26, 2010

Kalle Andreasson
"Vaccination against Her2/neu - expressing cancer using chimeric virus-like particles"
May 15, 2009

David Lindquist
"Studies on the occurence and effects of human papillomavirus in tumors of the head and neck"
March 28, 2008

Karin Tegerstedt
"Studies on polyomavirus virus-like particles : As vaccines and vectors for immune and gene therapy"
March 31, 2006

Liselotte Dahlgren
"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

Peter Priftakis
"Studies on human polyomavirus infection in immunosuppressed patients with polyoma related tumors"
October 12, 2001

Shirin Heidari
"Studies on persistant polyomavirus infection in relation to tumor development and options for vaccine and gene therapy"
June 1, 2001