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10125 - PD-L1+SOX10, Melanoma, TME

INTRODUCTION
Programmed death-ligand 1 (PD-L1) is a transmembrane protein that binds to the inhibitory receptor programmed death 1 receptor (PD-1), causing a down regulation of immune responses. PD-L1 is typically expressed on normal cells but has been observed in immune cells and tumor cells[1] while PD-1 is typically expressed on cytotoxic T-cells and other immune cells. Tumor cells can upregulate PD-L1 expression and avoid being attacked by the body’s immune system, making an interruption of the PD-1/PD-L1 interaction an attractive method for assisting the immune system in destroying tumor cells[2].

SOX10 is a nuclear transcription factor that has been shown to be a sensitive and specific marker for malignant melanoma[3]. Combining SOX10 and PD-L1 enables the user to assess whether PD-L1 expression originates from tumor cells or adjacent non-tumor cells.

This APP can be used to determine the number of tumor and non-tumor cells that express PD-L1 without having to outline tumor regions.

KEYWORDS
PD-L1, PD-1, T-cell, cancer, oncology, IHC, membrane, skin, melanoma, digital pathology, image analysis, B7-H1, SOX10, hematoxylin

METHODS
Based on an AEC red enhanced input band, SOX10 positive nuclei are classified using a combination of polynomial blob- and local linear filters. SOX10 negative nuclei are classified in the same manner but using a hematoxylin input band. Membranes are detected using two different scales of polynomial local linear filters on green/blue contrast and brown chromogen input bands. Postprocessing steps grow the nuclei blobs and combine/separate adjacent nuclei appropriately to match the input band. The classified PD-L1 positive membrane staining is verified by size, shape and intensity constraints and then applied to the adjacent cells to quantify the number of PD-L1 positive cells. An algorithm evaluates PD-L1 expression shared between SOX10 negative and positive cells to determine if tumor cells are truly PD-L1 positive or influenced by adjacent non-tumor cells.

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

  • Neg Stromal Cells: Number of SOX10-/PD-L1- cells
  • Neg Tumor Cells: Number of SOX10+/PD-L1- cells
  • PD-L1 Positive Stromal Cells: Number of SOX10-/PD-L1+ cells
  • PD-L1 Influenced Tumor Cells: Number of SOX10+ cells influenced by SOX10-/PD-L1+ cells
  • PD-L1 Positive Tumor Cells: Number of SOX10+/PD-L1+ cells
  • Total Stromal Cells: Sum of SOX10- cells
  • Total Tumor Cells: Sum of SOX10+ cells
  • PD-L1 Tumor Percentage: Percentage of SOX10+/PD-L1+ cells
  • PD-L1 Influenced Percentage: Percentage of SOX10+ cells influenced by SOX10-/PD-L1+ cells
  • PD-L1 Stromal Percentage: Percentage of SOX10-/PD-L1+ cells

AUXILIARY APPS
There are no Auxiliary APPs available.

WORKFLOW
Run the APP. Click the save button to transfer the results to the database.

STAINING PROTOCOL
There is no staining protocol available.

ADDITIONAL INFORMATION
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. AuthorTM offers 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 in collaboration with Aalborg University Hospital.

LITERATURE

  1. Zou, Weiping, and Lieping Chen. "Inhibitory B7-family molecules in the tumour microenvironment." Nature Reviews Immunology 8.6 (2008): 467-477.
  2. Pardoll, Drew M. "The blockade of immune checkpoints in cancer immunotherapy." Nature Reviews Cancer 12.4 (2012): 252-264.
  3. Willis, Brian C., et al. "SOX10: a useful marker for identifying metastatic melanoma in sentinel lymph nodes." Applied Immunohistochemistry & Molecular Morphology 23.2 (2015): 109-112.

 

RUO
FIGURE 1
FIGURE 1
Overview of a single PD-L1+SOX10 stained TMA core with melanoma tissue.
FIGURE 2
FIGURE 2
Region of the core on the original image. SOX10 positive nuclei are red and PD-L1 is brown.
FIGURE 3
FIGURE 3
Region of the core after PD-L1+SOX10 analysis with labels. SOX10+/PD-L1-: Pink, SOX10+/PD-L1+: Red, SOX10-/PD-L1-: Blue, SOX10-/PD-L1+: Purple, and SOX10+ influenced by SOX10-/PD-L1 in Orange.
FIGURE 4
FIGURE 4
Close-up of tissue area on the original image.
FIGURE 5
FIGURE 5
Same area as shown in Figure 4 after analysis with labels showing: SOX10+/PD-L1+: Red, SOX10-/PD-L1+: Purple, and SOX10+ influenced by SOX10-/PD-L1 in Orange.