Translational Research

Translational research translates basic scientific findings, new information or knowledge created into potential treatments for diseases. Its basic aim is to transfer knowledge from bench to bedside and to create new therapies, medical procedures, or diagnostics.

In the GANNET53 Project several partners are dedicated to translation research and provide substantial additional information on the effectiveness and efficacy of the drug combinations tested in the clinical trials.

The Translation Research activities in GANNET53 are conducted in three different work packages:

  • Biobaning
  • Companion Diagnostcs, and
  • Mouse Models and Human Xenografs


Biobanking

The biomaterials (samples of bodily fluid or tissue) collected in the GANNET53 TRIAL and the EUDARIO TRIAL form a comprehensive BIOBANK which helps researchers improve their understanding of the disease.

In GANNET53 the TOC-Network (http://www.toc-network.de), the largest European Biobank for ovarian cancer with tissues of more than 4.5006000 prospectively recruited patients, has taken over the task of biobanking. Within the TOC network, established and validated SOPs for biosampling and clinical data documentation are available and can be used by all centres.

The TOC Network is an initiative that started in 2004 at the Charité University in Berlin and is now including 162 clinical centres. The TOC Biobank has huge experience in coordinating the collection, processing, storage and transfer of human biological samples to promote biomedical research at the highest standard. Thus, using the TOC expertise, the GANNET53 consortium will be able to establish a high-quality biobank for archival and prospectively collected samples.

Objectives

  • Develop a database for biosample collection (virtual tumour bank) and biosample tracking
  • Organise prospective and retrospective collection, deposition and delivery of biomaterials
  • Provide Central Histopathological Review (CHR) to ensure the inclusion of solely Type II tumours into Phase II and to ensure tissue sample quality for experimental work

Description of Work

Results from the GANNET53 Clinical Trial

An outstanding biobank was established by the clinical partners from patients included in the randomised Phase II trial. Collected biomaterials include archival formalin-fixed, paraffin-embedded (FFPE) tumour tissues, biopsies of the actual relapse (fresh-frozen or FFPE), blood fractions (plasma, serum, cell pallets) collected taken at different time-points prior and during study treatment, circulating tumour cells (CTCs) in the blood, as well as ascites and pleural effusion samples.

Archival FFPE tumour tissues

Plasma, serum, cell pellets

Circulating tumour cells (CTCs)

Biopsies of the actual relapse

Ascites and pleura effusions


Companion Diagnostics

Clinical testing of new therapeutic drugs or drug combinations is very challenging. One very valuable and indispensable role within drug testing is more and more taken over by so-called Companion Diagnostics. Companion Diagnostics research tries to identify specific biological markers and provides important additional information that is essential for the safe and effective use of a corresponding therapeutic product.

Within the GANNET53 Project Companion Diagnostics play an important role as new molecular tests are being developed to predict and monitor the responsiveness to the tested drug combinations and to determine the exact p53 status of each patient.

Objectives:

  • Determine the exact p53 status in Phase II patients
  • Develope and evaluate a functional molecular test (proximity ligation assay) for its value to predict responsiveness to Ganetespib.
  • Performe CTC analyses before and during experimental therapy and evaluate it for its value to monitor responsiveness to Ganetespib.

Results achieved in the GANNET53 Clinical Trial:

Functional molecular test (proximity ligation assay) developed The proximity ligation assay (PLA) is an in situ technology, which allows a sensitive detection of protein, protein interactions. The signal can be localized with single molecule resolution. Specific antibodies recognize the two targets and are subsequently bound by PLA probes. They have a short DNA strand attached, and when in close proximity, they interact, are ligated and amplified via rolling circle amplification. Fluorochrome-labelled probes are then added to visualize the product, which is than seen as bright spot in the fluorescence microscope.

Read more

p53 mutational status in all Phase II patients determined The GANNET53 project is based on the hypothesis that mutant p53 is stabilized through the chaperon HSP90 and can therefore neither fulfil its function, nor be degraded. This stabilization is a prerequisite for gain-of-function capabilities promoting tumour growth. Furthermore, tumours carrying a missense TP53 mutation develop a dependency on the high protein levels and withdrawal should results in cytotoxicity. The HSP90 inhibitor Ganetespib is used to release stabilized p53 and target it for degradation. The GANNET53 trial includes patients with Type II ovarian cancer. This type of cancer is characterized by an almost ubiquitous presence of TP53 mutations. As part of the companion diagnostics, the TP53 mutation status of all patients is determined. For this purpose, it was mandatory that archival tissue of each patient included in the study was available. This analysis allows linking effects of the treatment to the presence of a TP53 mutation in general, and to certain mutations in specific. Furthermore, those results are the basis for additional projects which are conducted as part of the companion diagnostics, as for example the analysis of circulating DNA.

Read more


Mouse Models and Human Xenografts

Objectives

  • Provide the most stringent in vivo genetic and pharmacologic proof-of-principle for the underlying treatment concept in engineered mutp53 knock-in mice.
  • Stringently test the Ganetespib-mediated preferential killing of mutp53 cancer cells in human ovarian cancer models (cultured cells and nude mice).

Results

Publication: CELL DEATH AND DIFFERENTIATION (2016), 1–17: Strong antitumor synergy between DNA crosslinking and HSP90 inhibition causes massive premitoticDNA fragmentation in ovarian cancer cells. Kramer D, Stark N, Schulz-Heddergott R, Erytch N, Edmunds S, Roßmann L, Bastians H, Concin N, Moll UM, Dobbelstein M. doi: 10.1038/cdd.2016.124. [Epub ahead of print] PMID: 27834954; Link: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5299713/;