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Multiplex Quantitative Histologic Analysis of Human Breast Cancer Cell Signaling and Cell Fate

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Annual rept. 1 May 2007-30 Apr 2008

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Many molecular events and cellular processes are preserved in fixed human tumor specimens, and access to this wealth of information about human cancers in authentic context awaits a method for them to be quantified and analyzed. Some, such as cell signaling and cell fate decisions, are prognostically and therapeutically important, and can be revealed by immunohistological staining. We are developing a novel platform for immunohistological study of breast cancer specimens that will retrieve multiplex quantitative molecular information about tumor cells at a cytologic level. The platform will use multispectral microscopy to examine breast cancer specimens that have been immunostained for multiple structural and functional antigens using different chromogens and fluorophores. Staining for structural antigens nuclei, epithelial cytokeratins, E-cadherin, allows cells to be identified and classified as breast cancer cells. Staining for cell signaling and fate antigens p-ERK, p-AKT, Ki-67 reports on these important biological processes and events in cells. Multispectral microscopy permits staining for individual antigens to be distinguished and separated from staining for other antigens in multiplex-stained slides. The other component of the proposed platform is software for analyzing multispectral images individually and associatively. Algorithms will identify and classify cells in images, attribute signaling and cell fate events and processes to each cell, and reveal relationships among events through analysis of association among their stains.

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  • Anatomy and Physiology
  • Medicine and Medical Research

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