P53 Function Project

Background

p53 functional assays may be relevant to better understand consequences of p53/ATM mutations. Currently, within the ERIC framework different groups are using different techniques to assess p53 functionality but robustness, reproducibility and especially clinical value of each assay remain to be elucidated and need to be compared to each other.

Goal

  1. To compare different functional p53 assays and to correlate with FISH and sequencing
  2. To study prognostic impact of p53 function assays

Participants

Initially, the study will include the following ERIC members that are currently using the listed assays and techniques (between brackets, the contact person of each group).

  • Brno, Czech Republic: FASAY, mIR34a; ()
  • Salzburg, Austria: ()
  • Ulm/Heidelberg, Germany: miR34a, flow p53/p21, sequencing; ()
  • Paris, France: Flow p53/p21; Paris ()
  • Amsterdam, the Netherlands: MLPA; ()

At a later stage, other members can join.

Approach

  • First stage
    The amount of cells needed per group is approx. 20x106 CD5+/CD19+ cells. Some will need viable cells, some will need RNA (of =/- irradiated cells). As first step, Arnon/Eric will send around a vial of at least 20x106 viably frozen CLL cells with a purity of ≥95% of 3 patients to the different groups to study logistics, viability, cell number, overall feasibility, etc.
  • Second stage
    If this approach is feasible, all groups should send around 3 samples fulfilling the above criteria to the other centers. Of the total of 15 patient samples that will be tested, at least 5 should have a proven p53 mutation and if available 4-5 a proven ATM mutation.
    In case the first stage is not working, a different approach needs to be established, for instance only viable cells to centers that perform flow, who can send around tryzol specimens of non-irradiated and irradiated cells to centers who perform RNA/DNA analysis.
  • Third stage
    Depending on results from the second stage, a pilot can be run on interesting cases of a prospective trial (e.g. HOVON 68, CLL08, etc)

Time-line

First stage:  Mid 2011
Second stage- End 2011

Coordination

Arnon P. Kater, MD, PhD
Amsterdam, the Netherlands
E-mail: