Contact informationPlease contact Håvard E. Greger Danielsen for more information
The Cell-by-cell Sequential Multianalysis method (CCSM) is characterized by several analyses performed sequentially on the same tissue sample, with computerized cell-by-cell comparative analysis.
We consider the following questions:
To what extent are the observed changes in DNA amount and chromatin structure causally related, i.e. do quantitative alterations in DNA by necessity affect chromatin organization? To what extent are the observed changes in chromatin structure of a pure epigenetic nature? How do these changes relate to other established markers? Finally, how can we utilize the information to predict the outcome for the patients? To answer these questions, one must be able to analyze nuclei in single cells with respect to all of the selected changes and relate them to each other. This project seeks to do just that, by making cell-by-cell comparisons of the selected markers in the cells’ histopathological context.
A required analysis?
Most solid tumors are heterogeneous and only a subpopulation of cancer cells will develop the properties required to metastasize. Considering this, it is clear that analyses based on an average of all tumor cells are suboptimal and a cell-by-cell analysis and comparison are required for improved understanding of mechanisms involved in carcinogenesis and development of prognostic markers. A cell-by-cell analysis is a complex methodology, but we have developed most of the technology required for carrying out such studies.
Each cell an individual
The novelty of this project is that we are developing a high-resolution and high throughput method for cell-by-cell comparison of different markers. This is done by using a wide range of methods such as fluorescent and chromogenic in-situ hybridization (FISH and CISH), immunohistochemistry (IHC), DNA ploidy and Nucleotyping, all on the same cells in the same sections. The results are then combined and visualized on scans of standard HE-stained histopathological sections. This allows us to compare the results for the different markers within the same cells, concurrently, where the histological context and proximity of the different cells is known. This is in contrast to traditional marker studies, which have been based on molecular methods in which only average values for the samples are obtained.
The main objective of this project is to establish a system for cell-by-cell comparison of key markers in cancer, to produce high resolution Big Data and to relate the result to the clinical outcome for each patient as well as to different stages of cancer progression. Most of the necessary technical and methodological developments have already been established, while some minor technical adjustments, data production and data analysis remain. We will integrate data on DNA organization and chromatin structure with data collected on other relevant markers. Cell-by-cell comparison allows us to discover association undetectable by conventional methods. Therefore, we believe that we can contribute to the understanding of mechanisms behind changes in chromatin structure, genomic instability and the observed corresponding clinical outcome, in a novel way.