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Applying toponome imaging system for studying colon cancer

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Bhattacharya, Sayantan (2011) Applying toponome imaging system for studying colon cancer. MD thesis, University of Warwick.

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Abstract

Introduction
In a proof of principle study, we have applied an automated fluorescence toponome
imaging system (TIS) in order to examine, whether TIS can find protein network
structures distinguishing cancerous from normal colon tissue from same patient.
Methods/Materials
Cancer specimen and corresponding normal tissue were harvested at colectomy from a
single patient. 5μm sections were then prepared for TIS using a battery of different
antibodies, including a number of putative CSC markers. Expression of multiple protein
clusters was determined and Combinatorial Molecular Phenotypes (CMPs) were
analysed, using specific image-analysis tools.
Results
By using a three symbol code and a power of combinatorial molecular discrimination
(PCMD) of 221 per sub-cellular data point in one single tissue section, we demonstrate
an in situ protein network structure, visualized as a mosaic of 6,813 protein clusters
(Combinatorial molecular phenotype or CMPs) in the cancerous part of the colon. By
contrast, in the histologically normal colon, TIS identifies nearly 5 times the number of
protein clusters as compared to the cancerous part (32,009).
Discussion and Conclusion
By sub-cellular visualization procedures we found, that many cell surface membrane
molecules were closely associated with the cell cytoskeleton as unique CMPs in the
normal part of the colon, while the same molecules were disassembled in the cancerous
part, suggesting presence of dysfunctional cytoskeleton-membrane complexes. As
expected, glandular and stromal cell signatures were found, but interestingly also
potentially TIS signatures identifying a very restricted subset of cells expressing several
putative stem cell markers, all restricted to the cancerous tissue. The detection of these
signatures is based on the extreme searching depth, high degree of dimensionality, and sub-cellular resolution capacity of TIS. These findings provide the technological rationale
for the feasibility of a complete colon cancer toponome to be established by massive
parallel high throughput/high content TIS mapping.

Item Type: Thesis or Dissertation (MD)
Subjects: Q Science > QP Physiology
Library of Congress Subject Headings (LCSH): Colon (Anatomy) -- Cancer, Colon (Anatomy) -- Analysis, Colon (Anatomy) -- Imaging
Official Date: July 2011
Dates:
DateEvent
July 2011Submitted
Institution: University of Warwick
Theses Department: School of Life Sciences
Thesis Type: MD
Publication Status: Unpublished
Supervisor(s)/Advisor: Khan, Michael ; Mathew, George
Sponsors: Germany. Bundesministerium für Bildung und Forschung (BMBF) ; Deutsche Forschungsgemeinschaft (DFG) ; Klaus Tschira Stiftung ; Otto-von-Guericke-Universität Magdeburg. Fakultät der Medizin
Extent: xxii, 184 leaves : ill.
Language: eng

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