WP 09: Biomarker study


The objective of this WP is the creation of a common EuroHYP-1 Biobank that will be used for the study and discovery of a panel of biomarkers to biologically demonstrate therapeutic response to hypothermia. Thanks to the proposed biomarker substudy we might give for the first time in humans a powerful “proof of concept” of hypothermia. This substudy can also elucidate the more important mechanisms of benefit and neuroprotection associated to therapeutic cooling.
As a global objective we expect to identify “Efficacy Biomarkers” that will be markers that reflect BBB damage reduction or the decrease of inflammatory and brain damage related biomarkers; and “Safety Biomarkers” that will be some stroke outcome biomarkers aiding in the likely response to therapeutic cooling or development of hypothermia-related adverse events.

Work package Description

WP9 is led by Dr. Joan Montaner from VHIR-Barcelona. The work will be done in close collaboration with other academic centres such as Charité (Berlin) and also biomarker-related SMEs: Proteom Sciences and RANDOX.


1. The EUROHYP-1 Biobank will be established and hosted at the Hospital Vall d’Hebron Biobank Unit that follows best quality standards in compliance with national regulations regarding the usage of human biological samples (Biomedical Research Law 14/2007), and uses specifically designed software and infrastructure for sample management (Bio-e-Bank, VITROSOFT). We plan to standardize sampling protocols among centres in order to obtain serum, plasma and whole blood at different time-points (baseline, 1 day and 3 days) that will allow the consortium to study proteins, gene expression (mRNA) and polymorphisms (DNA) in the time-frame of the project and also in the future.


2. Testing BBB damage, inflammatory and brain damage biomarkers. VHIR-Barcelona will evaluate MMPs array including gelatinases (MMP-2 and MMP-9), collagenases (MMP-1, MMP-8, and MMP-13), stromelysines (MMP-3 and MMP-10), and MMP endogen inhibitors (TIMP-1 and TIMP-2), through (multiplexenzyme-linked immunosorbent assay [ELISA]) (SearchLight technology); Proteom Science will evaluate biomarkers: H-FABP, UFD1, RNABP, NDKA, GSTP-1 and Pro-BNP by standard ELISA and Randox will evaluate biomarkers from Cerebral Array I &II: BDNF, GFAP, NSE, NGAL, sTNFRI, D-dimer, TM and CRP using biochip analysers.


3. Testing biomarkers which can predict the risk of side-effects of cooling and long term outcome.Charité Berlin, based on data from previous own stroke trials will select and prove immune and stress markers as immediate early predictors for post-stroke infections and long term outcome. The remaining partners will select markers related to cardiac complications such as pro-BNP and D-dimer that have been involved in outcome mainly in cardioembolic strokes.


4. Discovery of new biomarkers using proteomics and gene arrays


5. Proposal of an optimal Point of Care (POC) prototype to analyze the selected biomarkers. All partners will analyze the best biomarkers coming form previous steps, and evaluate the feasibility of including those in aPOC to be used as safety and efficacy surrogate markers to guide stroke hypothermia.