Abstract C109: Quantitative analysis of multiple subtypes of immune system cells in cancer tissues
Authors: Krueger Joseph S. , Martin Nathan , Aeffner Famke , Milici Anthony , Alvarez John , Sharp Micheal 
Published: May 2016

Current cancer biology acknowledges the key role of the immune system in tumor biology, and promise for the modulation of immune system in cancer treatment. The composition of the inflammatory cell populations in tissues is reflective of the overall state of the Tumor Micro-Environment (TME), and the identification of distinct inflammatory cell types may hold prognostic or predictive value. Immunohistochemistry allows for reliable identification of the cell constituents to facilitate analysis of the TME while remaining in the tissue context.

Establishing a quantitative paradigm for inflammatory cell types and subtype profiling requires unbiased and automated whole-tissue based quantitation methods, which are capable of spatial integration of multiple inflammatory cell markers across the whole tissue. While single slide fluorescent multiplex approaches can address this need, the use of difficult-to-implement wet assay strategies involving multiplexing 6-8 fluorescent markers on the same tissue section are difficult to implement in a global clinical diagnostic lab setting. To answer this need, we combined novel advents in Tissue Image Analysis (TIA) to integrate spatial expression of serial-section stained whole tissue clinical lung cancer specimens.

In this proof-of-principle study,we were able to superimpose specific locations of individual cell types onto 6 serial sections and evaluate different inflammatory cell types. We used serial sections of clinical lung specimens stained for six immune phenotypic markers (CD68, CD4, CD8, CD33, FoxP3, and CD11b) to illustrate a repertoire of inflammatory cell types. Our proprietary CellMap algorithm was utilized to identify, enumerate, and determine the precise location of individual inflammatory cells in tissues on cell-by-cell basis in the tumor microenvironment (TME). Our proprietary FACTS (Feature Analysis on Consecutive Tissue Sections) approach was used to integrate the spatial expression of individual markers onto a reference H&E slide, and/or adjacent slides. Using the aligned FACTS data and our proprietary MultivariateMap approach, we integrated the patterns of each marker based on immune cell type function and their location relative to each other and the tumor epithelial cells.

In this study, we demonstrated how spatial integration of immune cell markers in the context of whole tissues can be applied to the diagnostic setting. By creating a comprehensive landscape of the immune system state in the tissue biopsies, we were able to identify crucial patterns which represent function and role in immune system biology. These approaches provide a robust platform for immuno-oncology applications by providing information on the state of the immune system in cancer using approaches implementable in the clinic. The use of these approaches will benefit further understanding of cancer pathology, and can directly lead to the development of diagnostic tests with clinical utility.

Citation Format: Joseph S. Krueger, Nathan Martin, Famke Aeffner, Anthony Milici, John Alvarez, Micheal Sharp. Quantitative analysis of multiple subtypes of immune system cells in cancer tissues. [abstract]. In: Proceedings of the AACR-NCI-EORTC International Conference: Molecular Targets and Cancer Therapeutics; 2015 Nov 5-9; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Ther 2015;14(12 Suppl 2):Abstract nr C109.