Flow Cytometry Assay for Tumor-Specific Proliferation Readout

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Solution Overview

Problem

Current diagnostic tests, such as OncotypeDx, struggle to distinguish between tumor cell proliferation and other cell types in breast cancer tumors, providing insufficient granularity for determining adjuvant chemotherapy, and have a lengthy turnaround time.

Innovation Solution

A cytometric assay that quantifies the percentage of tumor and normal cells expressing a cell proliferation marker by staining cells from a tumor sample, using flow cytometry to generate scatter plots and perform gating operations, allowing for a more granular proliferation readout and rapid results within one day.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Measurement precision

If current diagnostic tests like OncotypeDx are used to determine adjuvant chemotherapy, then a proliferation score can be obtained, but the test cannot distinguish between tumor cell proliferation and other cell types (such as immune cells), resulting in insufficient granularity

Engineering Contradiction:

Improvegranularity of proliferation readoutVSAvoidcomplexity of cell differentiation

Core Design Contradiction:

Measurement precisionVSDevice complexity

Solution Approach 1:

The assay segments the proliferation signal by using multiple markers: a proliferation marker (e.g., Ki-67) combined with either a tumor marker (e.g., cytokeratin) or a normal cell marker (e.g., CD3 for immune cells). This segmentation allows distinct identification of proliferating tumor cells versus proliferating normal/immune cells, resolving the granularity issue of conventional single-marker assays.

Inventive Principle:

Principle #1Segmentation

Solution Approach 2:

The patent introduces intermediary markers (tumor markers or normal cell markers) that act as mediators to classify the source of proliferation. These intermediary markers bridge the gap between the proliferation signal and cell identity, enabling precise attribution of proliferation to either tumor or normal cells through flow cytometry gating on double-positive populations.

Inventive Principle:

Principle #24Intermediary (Mediator)

2Productivity

If current diagnostic tests are used, then adjuvant chemotherapy decisions can be made, but the turnaround time is lengthy (two weeks)

Engineering Contradiction:

Improveturnaround timeVSAvoidgranularity of proliferation readout

Core Design Contradiction:

ProductivityVSMeasurement precision

Solution Approach 1:

The segmented dual-marker approach enables parallel processing of proliferation and cell type identification in a single flow cytometry run, eliminating sequential testing steps and reducing turnaround time while maintaining granular resolution of tumor versus normal cell proliferation.

Inventive Principle:

Principle #1Segmentation

Solution Approach 2:

The assay merges proliferation assessment and cell type identification into a single integrated flow cytometry experiment by simultaneously detecting multiple markers on the same cell population, thereby achieving both rapid turnaround and high measurement precision concurrently.

Inventive Principle:

Principle #5Merging (Combining)

3Loss of information

If a proliferation marker is measured without cell type differentiation, then the assay is simple, but the diagnostic information is insufficient for determining adjuvant chemotherapy

Engineering Contradiction:

Improvediagnostic information granularityVSAvoidmarker staining complexity

Core Design Contradiction:

Loss of informationVSDevice complexity

Solution Approach 1:

The assay segments the diagnostic information by combining proliferation marker data with cell type-specific marker data, creating distinct diagnostic categories for tumor cell proliferation versus normal cell proliferation. This segmentation recovers the lost diagnostic information without requiring complex additional testing.

Inventive Principle:

Principle #1Segmentation

Solution Approach 2:

The flow cytometry platform performs multiple functions simultaneously: it quantifies proliferation, identifies cell type, and provides diagnostic classification all in one assay, making the system universally applicable for both research and clinical diagnostic purposes while managing complexity through standardized multi-color flow protocols.

Inventive Principle:

Principle #6Universality (Multi-functionality)

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

The assay provides a more precise differentiation between tumor and normal cell proliferation, enabling better determination of adjuvant chemotherapy needs and reducing the turnaround time to one day.

Implementation Method 1

obtain flow cytometry data for the stained cells within each of the first and second aliquots

Methodology Applied

Scientific EffectFlow Cytometry:

Implementation Method 2

staining cells within a first aliquot derived from the sample for the presence of at least one cell proliferation marker

Methodology Applied

Scientific EffectFluorescence: Fluorescence

Data Source

PatentUS20260029324A1Proliferation assay for fixed solid tumors

Publication Date: 2026.01.29 VENTANA MEDICAL SYSTEMS INC

AI summary

Disclosed herein is a method of analyzing flow cytometry data for cells derived from homogenized whole tumor samples. In some embodiments, the present disclosure is directed to a cytometric assay for distinguishing between tumor cells expressing a cell proliferation marker and normal cells expressing the cell proliferation marker. In some embodiments, the present disclosure is also directed to quantifying a percentage of normal cells expressing a cell proliferation marker and a percentage of tumor cells expressing the cell proliferation marker.