This project aims for the development of an efficient immunotherapy for breast cancer, based on dendritic cell vaccines. Dendritic cells are the most promising target for a cell-based immunotherapy of cancer and several methods have been devised for successful loading of MHC molecules on dendritic cells with appropriate epitopes. These methods range from pulsing with peptides, proteins, cell lysate and RNA, through cross-priming with apoptotic cells, fusion with whole tumour cells or transfection with viral vectors. Each of these strategies has its advantages and it is still unclear how to recruite the multiple components of the immune system most effectively to eliminate tumour cells, a question this project is committed to answer.
The aim of this project is to develop and test an effective immunotherapy for the treatment of breast cancer based on dendritic cells (DC). This involves:
- generation of fully functional dendritic cells in a clinical scale;
- production of different types of immunogens based either on whole tumour information or based on MUC1 which is overexpressed and abberantly glycosylated in >90% of breast tumours;
- development of techniques for enhanced antigen transfer into dendritic cells;
- evaluation of the above immunogens in vitro using patients' T-cells (specific stimulation) and in mouse models (for tumour rejection);
- transfer of the most effective approach to GMP and design of a clinical trial.
DESCRIPTION OF THE WORK
The project is separated into three parts.
Generation of dendritic cells (DC)
An optimised protocol for the generation of dendritic cells in a clinical scale will be developed using methods and materials which allow a production to GMP standard. Different starting populations, cytokine combinations and culture conditions will be compared.
Production of four different types of immunogens
- To overcome the current limitation in the use of DC/tumour cell hybrids an efficient fusion process will be developed and compared with chemically induced fusion.
- To make whole-tumour RNA available for immunotherapy a clinically applicable protocol will be designed for in vitro amplification of tumour RNA isolated from breast cancer cells.
- Different viruses and virosome-like particles containing MUC1 DNA or MUC1 glycoproteins will be produced for transfection of DCs.
- 4. MUC1 glycopeptides based on the MUC1 tandem repeat carrying the cancer specific
T O-glycan have been shown by some of the partners to induce a strong proliferative T-cell response. To define a structure giving optimal T-cell response different MUC1 glycoproteins with defined glycyforms will be produced by recombinant CHO cells expressing different patterns of glycosyltransferases.
Evaluation of immune response
For the evaluation of their efficiacy, hybrids, RNA- or protein-loaded and virally transfected DCs will be compared in patients' T-cell stimulation and for tumour rejection in mouse models.
The most efficacious immunogen determined in the preclinical evaluation will be transferred to GMP and produced on a scale suitable for a clinical trial.
- generation of dendritic cells under clinically relevant conditions;
- production of hybrids from tumour and dendritic cells;
- amplification of RNA from breast cancer cells;
- production of viruses and virosome-like particles carrying MUC1 DNA or protein;
- modification of wild type CHO cells with recombinant glycosyltransferases;
- production of MUC1 glycoprotein with different cancer-related glycosylations;
- comparative evaluation of the above immunogens in vitro and in mouse models.
Dr Thomas Noll
Institute of Biotechnology 2
Forschungszentrum Jülich GmbH
52425 Jülich, Germany
Tel: +49-2461-613 955
Fax: +49-2461-613 870
Prof. Joyce Taylor-Papadimitriou
Dr. Joy Burchell
Breast Cancer Biology Group
Cancer Research UK
3rd Floor Thomas Guy House
SE1 9RT London, United Kingdom
Tel: +44-207-955 2310
Fax: +44-207-955- 2027
Prof. Jacques Bartholeyns
172 rue de Charonne
75011 Paris, France
Tel: +33-1-4009 0411
Fax: +33-1-4009 0425
Prof. Yvette van Kooyk
Molecular Cell Biology
v.d. Boechorstraat 7
1081 BT Amsterdam, The Netherlands
Tel: +31-20-444 8084
Fax: +31-20-444 8081
Dr Gerard Bos
Afd Inwendige Geneeskunde
Academisch Ziekenhuis Maastricht
6202 AZ Maastricht, The Netherlands
Tel: +31-43-3877 025
Fax: +31-43-3875 006
Prof. Karin Mölling
Institute of Medical Virology
University of Zürich
8028 Zürich, Switzerland
Tel: +41-1-634 2652
Fax: +41-1-634 4906
Prof. Henrik Clausen
Dental School, Glycobiology 24.5.33
University of Copenhagen
Norre Alle 20
2200 N Copenhagen, Denmark
Tel: +45-3532 6630
Fax: +45-3532 6835
Prof. Gunnar Hansson
Department of Medical Biochemistry
University of Gothenburg
413 90 Gothenburg, Sweden
Tel: +46-31-773 3488
Fax: +46-31-416 108