HER Imaging and Molecular Interaction Mapping in Breast Cancer
FP7 imagint
 

 Partner: ToposNomos Ltd

Project Leader
Walter Schubert, M.D., Assoc. Professor

Group members
Reference labs of ToposNomos Ltd

Collaborators
Molecular Pattern Recognition Research Group, University of Magdeburg, Germany


Selected References

Schubert, W.; Bonnekoh, B.; Pommer, A.J.; Philipsen, L.; Boeckelmann, R.; Maliykh, J.; Gollnick, H.; Friedenberger, M.; Bode, M.; Dress, A. Analyzing proteome topology and function by automated multidimensional fluorescence microscopy. Nat. Biotechnol. 2006, 24, 1270 – 1278. (Pubmed)

Friedenberger, M.; Bode, M.; Krusche, A.; Schubert, W. Fluorescence detection of protein clusters in individual cells and tissue sections by using toponome imaging system: sample preparation and measuring procedures. Nat Protoc. 2007, 2, 2285-2294. (Pubmed)

Schubert, W. A three symbol code for organized proteomes based on cyclical imaging of protein locations. Cytometry A 2007, 71, 352-360. (Pubmed)

Schubert, W, Gieseler, A., Krusche, A., Hillert, R. Toponome mapping in prostate cancer: detection of 2000 cell surface protein clusters in a single tissue section and cell type specific annotation by using a three symbol code. J Proteome Res. 8 (6), 2696-707 (2009). (Pubmed)

Schubert W. On the origin of cell functions encoded in the toponome. J Biotechnol. 149, 252-9 (2010). (Pubmed)
The human TOPONOME project: Translating the cellular protein network code (toponome) into efficient therapies

ToposNomos Ltd., Munich, Germany


Invention and introduction of the high-throughput robot imaging technology MELC/TIS toponomics allows to microscopically resolve large protein networks consisting of thousands of distinct protein clusters simultaneously at the cell surface; intracellularly; and transcellularly in one experiment. High-end TIS robots and specific software have been developed able to co-map protein clusters by automatically controlled repetitive cycles of fluorescence protein tagging, imaging and bleaching with only one dye thereby overcoming the spectral limitation of wavelengths, which is necessary to identify and decipher the high-dimensional combinatorial molecular structure of protein networks in a cell or in a tissue section. The resulting hierarchical organization of protein clusters uncovers lead proteins, that have been shown to control the integrity of protein networks. Blocking of lead proteins results in a disassembly of the network and loss of function in tumour cells, and similar proofs of principle were obtained by investigating neurological diseases substantiating that TIS toponomic imaging is a new way towards target discovery and predictive biomarkers in chronic diseases. The toponome project, launched by ToposNomos Ltd. is based on international patents. It aims at the deciphering of the protein network code in human cells and tissues in health and disease, by a cooperation of interdisciplinary institutions involving clinicians, molecular biologists, biochemists, computer scientists, and mathematicians.


Role of ToposNomos within the “Imagint Consortium”

Toponome imaging and identification of HER receptor protein network(s) by using TISTM technology





Feature: Toponome maps: (1) Nat Biotech 24, 1270, 2006; Cover: Over 7,000 Protein clusters in a single liver cell; (2) Corresponding Res. Highlight, Nature, 443, 609, 2006; (3) Nat Protoc. 2007;2(9):2285-94 Cover: Cell surface protein cluster network, peripheral blood T cell.

Figure: Breast cancer specific protein complex in situ. A cell in a breast cancer tissue (square section) was selectively imaged.
HER2/HER3 molecules indicated by arrows.

In breast cancers and their corresponding control tissues a typical so called CMP (combinatorial molecular phenotype) motif can be found; this was identified by using a tag library to co-map up to 72 different bio-molecules at a time.

The toponome analysis of cancerous and respective control tissue showed that these tissues have a CMP motif in common, encompassing three different lead proteins: a cell surface trans-membrane protein, a structural intracellular protein, and a signal transduction protein; and an assembly of additional proteins. Uniquely, cancerous tissue shows a specific new assembly with other proteins.


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