Projects

Laryngeal Papilloma 

Interaction between HPV, respiratory bacteria, and a dysfunctional local immune response may promote laryngeal papilloma. 

Although presence of human papillomavirus (HPV)-vaccines, there are new cases of HPV-diseases. Such a disease is laryngeal papilloma (LP) which is a benign tumor that can persist during life and is considered incurable, manifested by obstruction in the respiratory system, hoarseness, scarring after treatment. The incidence of LP is about 2/100,000, and Sweden has about 120 cases per year (1 of our patients had had 145 surgeries to be breathable). LP contains HPV6 or 11, but there is a lack of knowledge concerning interaction between HPV, respiratory bacteria, and a dysfunctional local immune response that may promote LP. 

Our aim is to increase the knowledge behind this local dysfunctional immune response against HPV. In  collaboration between ear-nose- and throat departments (ENT), microbiology and immunology laboratories we  will study clinical samples from LP-patients for presence of bacteria, repertoire of immune cells, and antibodies.  For LP-patients we will determine presence of viable pathogenic bacteria species as well as perform a deep analysis (technology, Nanopore) of the total genetic material of the microorganisms of LP, called the tumor microbiota. In addition, the immune status within the LP will be investigated by flow cytometry and by spatial technology (GeoMx) in order to characterize the precise sites of immune cell expressions in LP. We hope to increase the knowledge behind laryngeal papilloma with a long-term goal to eradicate this aero-digestive disease.

Leading researchers. Katarina Olofsson Umeå, Roland Rydell, Malin Lindstedt LTH, Ola Forslund

Main collaborators. Stefan Schwartz UU. Magnus Pålsson, LU, Aastha Sobti

Methylation assay to improve cervical cancer screening 

Early detection of methylation among HPV-positive women in cervical cancer screening.

Globally cervical cancer is a common disease with about 528,000 new cases and about 266,000 deaths cases (Year 2012). In Sweden about 500 cases and about 150 deaths occur yearly (NKCx Report, 2018). Cytological screening has facilitated early detection and treatment of pre-cancerous cervical lesions. Though, the sensitivity of a single cytology test is only around 60%. Since year 2014, the number of cervical cancer has increased with about 100 cases yearly in Sweden among women with normal cytology, indicating that the increase is related to factors that can be rectified. Today, several countries has introduced primary human papillomavirus (HPV) screening where cytology is most often used for triage of which women should be referred for immediate gynaecological follow up. Such approach is used in most regions in Sweden. 

Several hypermethylated promoters of tumor suppressor genes have been observed in many tumors and silencing allow cancer cells to escape the normal checkpoints of cell division and promote tumour growth Interestingly, DNA methylation patterns in cervical scrapings from women with cervical intraepithelial neoplasia (CIN) 2 and CIN3 are heterogeneous, with a subset displaying a cancer-like methylation-high pattern, suggestive for a higher cancer risk  Concerning cervical cancer, studies have indicated that promoter methylation of host cell genes such as FAM19A4 and mir124-2 increases with cervical disease severity. Also a DNA hypermethylation marker panel consisting of the six marker regions ASTN1, DLX1, ITGA4, RXFP3, SOX17, and ZNF671 has been associated with cervical cancer and its pre-cancerous stages. Overall methylation markers may be useful tools for triaging HPV-positive women in cervical cancer screening.  Potentially, a methylation assay could identify more severe lesions than cytology assessment among HPV-positive women who participate in cervical screening. In addition, methylation testing of HPV-positive vaginal self-samples may identify women with increased risk for development of severe lesions. 

In summary, we will study if methylation assays have the potential to pave the way to full molecular screening within the prevention-program of cervical cancer.

Leading researchers. Christer Borgfeldt, Ola Forslund

Main collaborators. Ylva Lindroth, Caroline Hellsten, Selamawit Mekuria

HPV-positive oropharyngeal carcinoma–Monitoring and early response evaluation using HPV-DNA in plasma (MERHPV)

HPV-DNA in plasma to characterize dynamics of HPV during treatment and early detection of residual disease after treatment.

HPV is the primary cause of an increase of oropharyngeal carcinoma (OPC). In Sweden a HPV prevalence of 73% was reported by us, where HPV16 predominates with a proportion of about 82% of HPV-positive cases. There is increased interest for detection of HPV-DNA in plasma which could be a valuable biomarker to identify ‘true’ residual disease after treatment of OPC. We will determine the value of HPV-DNA in plasma for treatment evaluation. For HPV-DNA assays in plasma, 150 patients with OPC will be invited during two years. In conjunction with diagnosis or staging, a biopsy is taken from the primary tumor and a plasma sample for the analysis of HPV-DNA prior to treatment. HPV-type identification from the primary tumor will be performed with an established method (Allplex), and quantification of the same HPV type in plasma from each patient using our semi-automated platform with DNA-extraction from plasma followed by quantitative in-house developed ddPCR, respectively. From each patient, weekly plasma samples, i.e., for 7 weeks, will be collected during the course of radiotherapy (RT), in order to study dynamics of HPV-load during treatment. For long time treatment evaluation and surveillance, plasma HPV-DNA will be analyzed every 3 months during the first two years and every six months year two-five after treatment. The study will most likely yield information if detection of HPV in plasma could improve monitoring after treatment.

Leading researchers. Johanna Sjövall, Maria Gebre-Medhin, Gabriel Adrian, Ola Forslund

Main collaborators.