banda transcan 2 2016ul

Title: Automated, absolute quantification of MRD in ALL patients by droplet microfluidics, single DNA copy barcoding and IG/TR amplicon NGS

Project Coordinator:
Tobias Hutzenlaub (Germany), University of Freiburg, Department of Microsystems Engineering – IMTEK Lab for MEMS Applications, Freiburg

Project Partners:
Michaela Kotrova (Germany), University Hospital Schleswig-Holstein, Unit for Hematological diagnostics, Kiel
Anastasia Hadzidimitriou (Greece), Centre for Research and Technology Hellas (CERTH), Institute of Applied Biosciences (INAB), Thessaloniki
Giovanni Cazzaniga (Italy), Fondazione Matilde Tettamanti e Menotti de Marchi ONLUS (Fondazione Tettamantii), Monza
Alessandro Rambaldi (Italy), ASST Papa Giovanni XXIII, Department of Oncology and Haematology - Hematology Unit, Bergamo

Project Abstract:
The assessment of minimal residual disease (MRD) during first-line therapy is regarded as most important prognostic factor in adult and childhood acute lymphoblastic leukemia (ALL). However, current methods used for MRD assessment have major drawbacks. Multiparameter flow cytometry is not fully standardized and requires expert skills. Allele specific PCR of clonal immunoglobuline (IG) and T-cell receptor (TR) gene rearrangements is time consuming and needs patient-specific reagents, thus making it ineligible for IVD-guided analytical validation. In this context amplicon based IG/TR NGS was introduced, which allows for the characterization of millions of IG/TR rearrangements in parallel without the need for patient specific reagents, thus allowing IVD-guided analytical validation. The downside of this method is that it is prone to amplification biases that hamper correct quantification. Also, contamination of libraries leading to false positive results constitutes a severe problem in diagnostic routine. The goal and hypothesis of Quant-ALL is to eliminate these disadvantages by a combination of innovative high throughput molecular tools.
Aims and methods: (1) Establishment of an amplicon based IG/TR NGS approach using single DNA copy barcoding and droplet microfluidics for IG/TR based MRD quantification in ALL. (2) Optimization of existing bioinformatics tools used for IG/TR NGS to quantify index IG/TR sequences. (3) Transfer of this technology to a pre-existing centrifugal microfluidic platform for full automation and standardization. (4) Validation of technology for childhood and adult ALL using biobanked follow-up samples that are MRD quantified with standard technologies.
Expected results are the full automation of a high throughput technology for MRD quantification in ALL. This technology will be standardized and validated to allow a wide distribution to ALL MRD reference laboratories and fundamentally improve MRD quantification in childhood and adult ALL.


(Project funded under JTC 2017)


Launch of the call: JTC 2021
The new TRANSCAN-3 project consortium is launching the first Joint Transnational Call for proposals, co-funded by the European Commission, on:
"Next generation cancer immunotherapy: targeting the tumour microenvironment"

International Networking Event (virtual)
on 22 April 2021 hrs 9:00 CET
will focus on the forthcoming co-funded JTC 2021 on:
"Next generation cancer immunotherapy: targeting the tumour microenvironment" 
the event is organized by TUBITAK under the “Turkey in Horizon 2020 Phase II” Technical Assistance Project
in cooperation with TRANSCAN-3

 Sustained collaboration of national and regional programmes in cancer research
is the new project funded by the European Commission under H2020  which started on 1st March 2021 and will last five years (G. A. no. 964264)

A TRANSCAN-2 brochure, highlighting key achievements, is available for download


eu flagship

This project has received funding from the European Union’s Horizon 2020 research and innovation programme under grant agreement No 643638.

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