Clinical trial NCT02207686: Assessment of Residual VHL Function in Tumors - Can it Predict the Patients' Individual Course of Disease?
Official title: Assessment of Residual VHL Function in Tumors - Can it Predict the Patients' Individual Course of Disease?
Study type: OBSERVATIONAL
Overall status: UNKNOWN
Conditions: Von Hippel-Lindaus Disease
Keywords: CNS hemangioblastoma
Renal Cell Carcinoma
VHL germline mutation
VHL somatic mutation
Tumor development
Lead sponsor: University of Copenhagen
Sponsor class: OTHER
First posted: 2014-08-04
Enrollment: 50
Sex: ALL
Minimum age (years): 18
Standard ages: ADULT
OLDER_ADULT
Accepts healthy volunteers: no
Observational model: COHORT
Time perspective: RETROSPECTIVE
Sampling method: NON_PROBABILITY_SAMPLE
Study population: vHL patients over 18 years of age who have had at least one vHL-related tumor removed, and for whom a reference DNA sample (from blood or normal tissue) and tumor tissue (paraffin-embedded or fresh frozen) can be obtained.

Brief summary: The investigators aim to analyze tumors from vHL patients who have different courses of disease and different types of VHL gene alterations to characterize which types of genetic alterations the tumors contain and how these alterations affect the tumor cells' behavior on a molecular level. The investigators will then compare these observations to vHL disease outcome in patients and families.

It is already known that most vHL tumors develop when both copies of the VHL gene in a cell are inactivated. The first copy is inactivated in all the person's cells from birth ("first hit"), leaving just one functional copy. A tumor can develop from cells where the second copy is also inactivated ("second hit"). So far, only the molecular consequences of the first hit have been investigated. It is our hypothesis that both the first and second hits in combination have consequences for tumor development and clinical outcome. The investigators will include tumors from patients with different disease courses and different types of "first hits" and analyse the tumors' DNA in order to find correlations between the first and second hits and patients' and families' medical histories. The investigators hereby hope to give new insights into how vHL tumors grow and which genetic factors influence tumor development. These results will contribute to the current knowledge of vHL and help us get one step closer to be able to predict an individual's tumor risks and need for surveillance.

Detailed description: In vHL tumors from patients with different phenotypes and different genetic backgrounds, the investigators aim to assess both the nature of germline and somatic VHL mutations along with the total residual VHL protein (pVHL) activity in tumor cells and evaluate association to disease outcome in patients and families.

vHL tumor development follows Knudson's "two-hit-mode": patients are born with a germline mutation in one copy of their VHL gene in all the cells of the body - "the first hit". Somatic mutation in the other copy of the VHL gene - "the second hit" - initiates tumor development. Pheno-genotype correlations are well known in vHL, but have so far been explained exclusively by the nature of the germline mutation (the first hit). It is the investigators' hypothesis that it is the total residual activity of the protein (pVHL) present in the tumor after both first and second hit, which decides each tumor's destiny.

The investigators will apply Sanger sequencing, MLPA (and multiplex ligation dependent probe amplification), LOH (Loss of heterogeneity) analysis, methylation assays, FISH (Fluorescence In Situ Hybridization), and immunohistochemistry to characterize the germline and somatic mutations, determine presence of mRNA and pVHL, and assess residual VHL activity in each tumor. All these methods are already established in our laboratory.

The results will give deeper insight to vHL tumorigenesis and pave the road to future individual prediction of each patient´s course of disease and need for surveillance.

Eligibility criteria: Inclusion Criteria:

* vHL diagnosed in patient
* Patient over 18 years of age
* Informed consent to participate can be obtained
* Patient has had at least one vHL-related tumor removed
* A reference DNA sample (from blood or normal tissue) and tumor tissue (paraffin-embedded or fresh frozen) can be obtained.

Exclusion Criteria:

* Patients under the age of 18 years
* Patients who had not previously had a vHL-related tumor removed
* Patients whos previously removed tumor tissue cannot be obtained or is of such a quantity or quality that no exact histological analysis can be done and/or no DNA can be extracted

Primary outcomes:
measure: Residual pVHL activity measured by amount of VHL mRNA in tumor cells
timeFrame: Two years
Secondary outcomes:
measure: Presence of VHL protein (pVHL) in tumor cells
timeFrame: Two years
measure: Type of second hit (somatic mutation) found in DNA from tumor cells
timeFrame: One year
measure: Patient's age at tumor diagnosis
timeFrame: Two years
measure: Patient's total tumor burden
timeFrame: Two years
Other outcomes:
measure: Type of clinical vHL in the patient's family (e.g. type 1, type 2)
timeFrame: Two years
Locations:
city: Copenhagen
country: Denmark
facility: Department of Cellular and Molecular Medicine, University of Copenhagen
state: Copenhagen N
Study officials:
affiliation: Department of Cellular and Molecular Medicine, University of Copenhagen
name: Marie Luise Bisgaard, MD
role: PRINCIPAL_INVESTIGATOR
References: