Multiplexed Immunofluorescence Imaging for VISTA Expression Scoring

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Solution Overview

Problem

Current immunotherapy approaches face challenges in effectively predicting and enhancing therapeutic responses due to the variability of VISTA expression in the tumor microenvironment, necessitating improved methods for characterizing immunoregulatory protein expression in individual immune cells.

Innovation Solution

A multiplexed method of multispectral immunofluorescence imaging is developed to simultaneously detect VISTA and other immune checkpoint proteins along with specific markers for various cell types in tumor tissue samples, allowing for detailed quantification and scoring of VISTA expression.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Quantity of substance

If traditional single-marker immunohistochemistry is used to detect immune checkpoint proteins, then the detection process is simple, but multiple markers cannot be detected simultaneously and the understanding of tumor microenvironment remains limited

Engineering Contradiction:
Improvenumber of markers detectedVSAvoidimaging method complexity
Core Design Contradiction:
Quantity of substanceVSDevice complexity

Solution Approach 1:

The patent combines multiple immunohistochemistry staining procedures into a single multiplexed imaging assay that detects VISTA, PD-L1, and other immune checkpoint proteins simultaneously in the same tissue section, allowing comprehensive characterization of the tumor microenvironment without processing multiple separate samples

Inventive Principle:
Principle #5Merging (Combining)

Solution Approach 2:

The multiplexed imaging system serves multiple functions: it detects various immune checkpoint proteins (VISTA, PD-L1, CTLA-4), identifies different cell types (tumor cells, immune cells), and provides spatial information about their interactions within the tumor microenvironment, all through a single integrated platform

Inventive Principle:
Principle #6Universality (Multi-functionality)

2Measurement precision

If VISTA expression is assessed without cell-type specificity, then the assessment is straightforward, but the variability of VISTA expression across different cell types cannot be characterized

Engineering Contradiction:
ImproveVISTA expression characterizationVSAvoidscoring method complexity
Core Design Contradiction:
Measurement precisionVSDevice complexity

Solution Approach 1:

The patent applies different detection and scoring approaches to different cell types within the tumor microenvironment. VISTA expression is specifically quantified in tumor cells versus immune cells separately, and the spatial relationships between VISTA-positive cells and other immune checkpoint proteins are analyzed to provide localized functional insights

Inventive Principle:
Principle #3Local quality

Solution Approach 2:

The system creates digital copies and overlays of the tissue section stained with different markers, allowing virtual separation and analysis of VISTA expression in specific cell populations without physically separating the cells, thereby maintaining spatial context while achieving cell-type-specific measurement

Inventive Principle:
Principle #26Copying

3Reliability

If immunotherapy response prediction relies on limited markers, then the predictive model is simple, but the ability to predict and enhance therapeutic response is insufficient

Engineering Contradiction:
Improvetherapeutic response predictionVSAvoidnumber of markers required
Core Design Contradiction:
ReliabilityVSQuantity of substance

Solution Approach 1:

The patent performs preliminary comprehensive characterization of the tumor microenvironment by detecting multiple immune checkpoint proteins (VISTA, PD-L1, CTLA-4) and their spatial relationships before initiating or adjusting immunotherapy treatment, allowing clinicians to predict response likelihood and select optimal therapeutic combinations in advance

Inventive Principle:
Principle #10Preliminary action

Solution Approach 2:

The detailed multiplexed imaging data provides feedback about the functional state of the tumor microenvironment, including the presence and interaction of different immune checkpoint proteins, which can guide decisions about whether to proceed with immunotherapy, combine multiple checkpoint inhibitors, or modify treatment based on the observed immune landscape

Inventive Principle:
Principle #23Feedback

Applied Scientific Principles

This section explains which scientific principles are used to turn an abstract innovation direction into a practical engineering solution.

Function Achieved in This Case

This method enables robust characterization of VISTA expression across different cell types within the tumor microenvironment, providing valuable insights for predicting treatment responses and identifying potential therapeutic targets.

Implementation Method 1

contacting the tissue sample with a primary antibody directed to VISTA

Methodology Applied
Scientific EffectAntibody-antigen binding:

Implementation Method 2

visualising each of the antibodies in the tissue sample with a reagent that generates a detectable signal corresponding to each of the antibodies

Methodology Applied
Scientific EffectFluorescence: Fluorescence

Data Source

PatentEP4560319A1Immune and tumour cells expression of vista in a panel of cancer indications
Publication Date: 2025.05.28 PIERRE FABRE MEDICAMENT SAS
  • EP4560319A1 patent drawingFigure 1
  • EP4560319A1 patent drawingFigure 2~3
  • EP4560319A1 patent drawingFigure 4~5

AI summary

The present invention provides multiplex assays for improved scoring of tumour tissues stained with an immune checkpoint protein such as VISTA. Methods of ranking multiple tumour tissues and of diagnosing cancer patients are also provided. The methods disclosed are also useful for identifying patient responsive to anticancer therapy.