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10132 - m-PGES1, Melanoma, TME

INTRODUCTION

Microsomal prostaglandin E synthase-1 (mPGES1) (genetic determinant of early onset breast cancer and predisposition to cancers of the ovary, colon, prostate gland and Fallopian tubes) is the downstream enzyme in the prostaglandin pathway. Expression of mPGES1 is inducible in conditions of inflammation and hypoxia and its expression is regulated by a number of transcriptional factors. Clinical studies have shown increased levels of mPGES-1, associated with increased PGE2 production, within a number of human cancers, including colon, lung, stomach, pancreas, cervix, prostate, papillary thyroid carcinoma, head and neck squamous carcinoma, brain tumors and melanoma.

The "10132 - m-PGES1, Melanoma, TME" APP detects nuclei and classifies them as either negative, 1+, 2+ or 3+ based on the m-PGES1 staining expression present in each nucleus' vicinity.


KEYWORDS
m-PGES1, microsomal prostaglandin E synthase-1, melanoma, skin, cancer, oncology, IHC, tumor micro environment


METHODS
To identify the nuclei, the APP performs a two-stage polynomial blob filtering on a blue-enhanced feature image and delimits them using local linear filtering. Each pixel with DAB staining is classified as low, mid and high based on the intensity and grouped together locally. Each nucleus is then classified based on its surroundings in the order of 3+, 2+, 1+ and negative to emphasize the strongest staining present in each nucleus’ vicinity.


QUANTITATIVE OUTPUT VARIABLES
The output variables obtained from this protocol include:

  • Number and percentage of negative, 1+, 2+, and 3+ cells in both stromal and tumor tissue
  • H-score = (% of 3+)x3 + (% of 2+)x2 + (% of 1+) for both stromal and tumor tissue

AUXILIARY APPs
APP: “01 Detect TumorStroma”
The auxiliary APP: “01 Detect TumorStroma” is used for automatic tumor and stromal tissue detection. The analysis APP will then provide results for tumor and stromal tissue separately.

WORKFLOW
Step 1: Load and run the APP “01 Detect TumorStroma” for tumor and stromal tissue identification. Maually correct the result if necessary.
Step 2: Load and run the APP “02 m-PGES1 Analysis” for the quantification of cells.

STAINING PROTOCOL
There is no staining protocol available.

ADDITIONAL INFORMATION
The APP utilizes the Visiopharm Engine™ and Viewer software modules, where Engine™ offers an execution platform to expand processing capability and speed of image analysis. The Viewer allows 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 Author™ module the APP can be customized to fit other purposes. Author™ offers a comprehensive and dedicated set of tools for creating new fit-for-purpose analysis APPs, and no programming experience is required.

REFERENCES
There are no references available.

RUO
Figure 1
Figure 1
Nuclei surrounded by m-PGES1 negative and weakly positive staining.
Figure 2
Figure 2
Nuclei classified as negative (blue) or 1+ (yellow) based on the m-PGES1 staining of each nucleus' surroundings.
Figure 3
Figure 3
Nuclei surrounded by m-PGES1 intermediate and strongly positive staining.
Figure 4
Figure 4
Nuclei classified as 2+ (orange) or 3+ (red) based on the MIF staining of each nucleus' surroundings.
Figure 5
Figure 5
Sample with stromal and tumor tissue.
Figure 6
Figure 6
Tumor and stroma identification by auxiliary APP. Stroma regions are outlined by a red region of interest (ROI) and tumor regions by a blue ROI. Quantitative output variables such as H-score is given for stroma and tumor region individually.