Optimization of a standard protocol for linear amplification of tumor RNA extracted from Fine Needle Aspirates (FNA) of pancreatic tumors.

Pancreatic cancer (PC) has a very poor prognosis despite conventional
treatment. Therefore, development of new strategies to treat this aggressive
form of cancer is urgently needed. In the past decades a novel approach has
been considered to be applicable to cancer therapy: Immunotherapy using
Dendritic Cell (DC) vaccines. A few years ago a new research group has been
established at the AMC in collaboration with Sanquin. For the last years this
group is occupied with the development of a vaccine where DCs are pulsed with
autologous total tumour RNA. In the future, these vaccines will be used to
induce a strong Cytotoxic T-cell (CTL) response specifically directed against
the patients own tumour cells. In the future phase 1 clinical trial we want to
administer a maximum of 10 vaccines. These vaccines mainly consist of
autologous patient material and in order to obtain this material (monocytes and
cancer cells) patients need to undergo invasive and time consuming procedures
such as endoscopies and leukapheresis procedures. In order to subject patients
to as less invasive- and time consuming procedures as possible, our aim is to
produce multiple vaccines at once. The total tumor RNA is obtained from FNA's
of the patient's tumor, but due to limited access to pancreatic tumors, the
amount of RNA obtained in this way is only sufficient for the production of a
single vaccine. To obtain a sufficient amount of RNA for the production of
multiple vaccines, the RNA obtained from FNA*s will be linearly amplified. In
this study the protocol for linear amplification of total tumor RNA will be
optimized.

Optimize the protocol for linear amplification of tumour RNA extracted from
FNA's under GMP conditions.

It concerns an observational study with PC patients who are primarily being
referred to the AMC for staging and treatment. A total of thirty patients will
be included at the outpatient clinic after receiving informed consent and
signing written informed consent. Patients will undergo a procedure where
patient material will be obtained for this study. This procedure will be a
routine procedure performed at the AMC, where the patients tumour will be
staged and material will be taken for histological diagnosis. During this
procedure we will take extra cytologic material that we will use to optimize
our protocol for the linear amplification of total tumor RNA.

Time: The EUS will be prolonged with a maximum of five minutes due to obtaining
extra material for the study. Interventions: During the planned EUS extra FNA's
will be obtained. There's a scarce chance of bleeding from the aspiration site
after this procedure and there is a very small chance of infection due to the
FNA.