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Tissue-print and print-phoresis as platform technologies for the molecular analysis of human surgical specimens: mapping tumor invasion of the prostate capsule

Abstract

Molecular profiling of human biopsies and surgical specimens is frequently complicated by their inherent biological heterogeneity and by the need to conserve tissue for clinical diagnosis. We have developed a set of novel 'tissue print' and 'print-phoresis' technologies to facilitate tissue and tumor-marker profiling under these circumstances. Tissue printing transfers cells and extracellular matrix components from a tissue surface onto nitrocellulose membranes, generating a two-dimensional anatomical image on which molecular markers can be visualized by specific protein and RNA- and DNA-detection techniques. Print-phoresis is a complementary new electrophoresis method in which thin strips from the print are subjected to polyacrylamide gel electrophoresis, providing a straightforward interface between the tissue-print image and gel-based proteomic techniques. Here we have utilized these technologies to identify and characterize markers of tumor invasion of the prostate capsule, an event generally not apparent to the naked eye that may result in tumor at the surgical margins ('positive margins'). We have also shown that tissue-print technologies can provide a general platform for the generation of marker maps that can be superimposed directly onto histopathological and radiological images, permitting molecular identification and classification of individual malignant lesions.

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Figure 1: Collection of tissue prints from prostate specimens.
Figure 2: Tissue prints peel a microscopic layer of cells from the surface of the tissue specimen.
Figure 3: Mapping markers of extracellular matrix turnover associated with tumor invasion of the prostate capsule.
Figure 4: Tumor infiltration of the prostate capsule colocalizes with focal concentrations of collagen fragments and MMP activity on the external surface of the prostate gland.

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Acknowledgements

The work described in this proposal was supported by a supplement to US National Institutes of Health grant CA86365 to SMG and a Research Grant from General Electric Medical Systems to REL.

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Correspondence to Sandra M Gaston.

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Supplementary information

Supplementary Fig. 1

Reverse transcriptase PCR (rtPCR) of PSA mRNA from an interior prostate tissue print. (PDF 47 kb)

Supplementary Fig. 2

Quantitative rtPCR profiles of β2M, PSA and AMACR transcripts in RNA extracted from tissue prints of 16 prostate needle biopsies. (PDF 212 kb)

Supplementary Fig. 3

Detection of protein markers by print-phoresis. (PDF 71 kb)

Supplementary Fig. 4

Flow chart of spatial-molecular profiling of tumor-associated proteins with tissue print and print-phoresis technologies. (PDF 140 kb)

Supplementary Fig. 5

Print-phoresis profiling of collagen fragments colocalized with a high grade tumor focus within the parenchyma of the prostate gland. (PDF 122 kb)

Supplementary Fig. 6

Print-phoresis profiling of collagen fragments on the external surface of the prostate capsule. (PDF 227 kb)

Supplementary Fig. 7

Histopathologically evident tumor invasion of the prostate capsule. (PDF 101 kb)

Supplementary Table 1

Tissue print marker-marker associations on the external surface of radical prostatectomy specimens: 78 sextants (PDF 22 kb)

Supplementary Methods (PDF 29 kb)

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Gaston, S., Soares, M., Siddiqui, M. et al. Tissue-print and print-phoresis as platform technologies for the molecular analysis of human surgical specimens: mapping tumor invasion of the prostate capsule. Nat Med 11, 95–101 (2005). https://doi.org/10.1038/nm1169

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