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10015 - TRIM29, Bladder Cancer

INTRODUCTION

Tripartite motif-containing 29 (TRIM29) is a member of the TRIM protein family. TRIM29 expression has been shown to increase cell proliferation through the inhibition of p53 activity. TRIM29 binds p53, resulting in its export out of the nucleus, and this interaction is fine-tuned by post-translational acetylation of TRIM29 [See REFERENCES: 1]. It has been suggested that TRIM29 is involved in the response to DNA damage through its regulation of p53 and that it could potentially function as an oncogene that promotes tumor growth [See REFERENCES: 2]. Immunohistochemical analysis has showed that the expression of TRIM29 is increased in most invasive pancreatic cancers and is correlated with high expression levels of β catenin [See REFERENCES: 3]. A clinical study in gastric cancer has also shown that TRIM29 expression correlates with poor histological grade, large tumor size, extensive tumor invasion and lymph node metastasis [See REFERENCES: 4]. Recently, TRIM29 has been shown to function as a tumor suppressor in breast cancer [See REFERENCES: 5].

This APP can be used for quantifying the cytoplasmic expression of TRIM 29. A very high correlation to manual scoring, in a study involving 300 patients (R2 = 0.78), was demonstrated (publication pending).

KEYWORDS

TRIM 29, bladder cancer, metastasis, immunohistochemistry, image analysis, quantitative, digital pathology.

METHODS

The method used for computing the TRIM 29 expression is based on the color deconvolution of Hematoxylin, isolating all the nuclei within the tumor region [See EXAMPLES: 2]. A second color deconvolution is used to isolate DAB within the tumor region [See EXAMPLES: 3]. The DAB intensity threshold is used to determine positivity versus negativity of the cytoplasmic expression. Note that both color deconvolution bands have been smoothed to reduce high-frequency noise, including the blocking that originates from the JPEG compression of the digital slide. 

From the segmentation, the average DAB intensity across all positive cytoplasm areas is measured and subtracted from 255. This is done to associate high values with a high staining intensity, which is more intuitive.

It should be noted that this approach to quantifying biomarker expression is vulnerable to variations in protocols for tissue preparation (fixation time, tissue thickness, reagents, auto-stainer type and settings, etc.). Furthermore, the application of this APP for TMA study designs is robust within the TMA. 

This APP requires manual outlining of tumor regions

QUANTITATIVE OUTPUT VARIABLES

The output variables obtained from this protocol include:

- ROI Area: Total area of the outlined tumor ROI

- Cytoplasmic Area = ROI Area - Area of Nuclear Profiles

- Positive Cytoplasmic Area: Area within ROI with DAB intensity above defined threshold

- Positive Area Fraction = Positive Cytoplasmic Area / Cytoplasmic Area

- Cyt Intensity = 255 - average DAB intensity for Positive Cytoplasmic Area

AUXILIARY APPS (included)

Auxiliary APPs are used for additional process steps, e.g. finding Region of Interest (ROI).

There are no Auxiliary APPs available.

STAINING PROTOCOL

There is no staining protocol available

ADDITIONAL INFORMATION


This APP was developed for, and validated by, Niels Fristrup MD and Dr. Lars Dyrskjøt Andersen, Center for Molecular Clinical Cancer Research Department of Molecular Medicine (MOMA) Aarhus University Hospital

The APP utilizes the EngineTM and Viewer software modules, where EngineTM offers an execution platform to expand processing capability and speed of image analysis. Viewer gives a fast review together with several types of image adjustment properties ex. outlining of regions, annotations and direct measures of distance, curve length, radius, etc. 
By adding the AuthorTM module the APP can be customized to fit other purposes. AuthorTMoffers a comprehensive and dedicated set of tools for creating new fit-for-purpose analysis APPs, and no programming experience is required.

REFERENCES

USERS


This APP was developed for, and validated by, Niels Fristrup MD and Dr. Lars Dyrskjøt Andersen, Center for Molecular Clinical Cancer Research Department of Molecular Medicine (MOMA) Aarhus University Hospital

LITTERATURE

1. The ATDC (TRIM29) protein binds p53 and antagonizes p53 mediated functions, Z. Yuan et al., Mol. Cell. Biol. 30, 3004-3015, 2010.

2. TRIM proteins and cancer, S. Hatakeyama, Nat Rev Cancer Oct 7;11(11):792-804, 2011. doi: 10.1038/nrc3139.

3. Oncogenic function of ATDC in pancreatic cancer through Wnt pathway activation and β-catenin stabilization, L. Wang et al., Cancer Cell 15, 207-219, 2009.

4. Tripartite motif-containing 29 (TRIM29) is a novel marker for lymph node metastasis in gastric cancer, Y. Kosaka et al., Ann. Surg. Oncol. 14, 2543-2549, 2007.

5. TRIM29 Functions as a Tumor Suppressor in Nontumorigenic Breast Cells and Invasive ER+ Breast Cancer, J. Liu, B. Welm, K. M. Boucher, M. T. Ebbert, P. S. Bernard, Am J Pathol. Dec 2, 2011

RUO
FIGURE 1
FIGURE 1
Field of view showing a tumor region in an image of a bladder tissue TMA core stained by IHC for TRIM29.
FIGURE 2
FIGURE 2
The same field of view as in Figure 1 after automatic color deconvolution isolating the Hematoxylin staining, and thereby highlighting all cell nuclei.
FIGURE 3
FIGURE 3
The same field of view as in Figure 1 after automatic color deconvolution isolating the DAB staining, and thereby highlighting the TRIM29-positive cytoplasm.
FIGURE 4
FIGURE 4
The same field of view as in Figure 1 after automatic segmentation of all nuclei (blue label) and intense TRIM29-positive cytoplasm (red label).