Ex Vivo Apoptosis Screening for Hematological Neoplasms

Resolve Bottlenecks,
Find Innovative Solutions
Generate Solutions

Solution Overview

Problem

Current personalized medicine tests for cancer patients are limited by long incubation times, inability to distinguish between drug effects on tumor cells and normal cells, and limited ability to evaluate multiple drug combinations, which restricts their clinical usefulness and effectiveness.

Innovation Solution

A cell-based screening platform using automated sample preparation and flow cytometry that allows for rapid analysis of drug sensitivity profiles by measuring apoptosis in whole blood or bone marrow samples, enabling evaluation of multiple drug combinations and concentrations within 24 to 72 hours, and identifying effective polytherapy regimens.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Reliability

If traditional ITRT methods are used to evaluate drug sensitivity, then clinical correlation is achieved, but incubation time is extended to weeks rather than days

Engineering Contradiction:
Improveclinical correlationVSAvoidincubation time
Core Design Contradiction:
ReliabilityVSLoss of time

Solution Approach 1:

The patent changes the measurement parameter from total cell death to apoptosis-specific markers (caspase activation, phosphatidylserine externalization) detectable by flow cytometry. This parameter change enables differentiation between apoptotic and non-apoptotic cell death, allowing for shorter incubation periods while maintaining clinical correlation because apoptosis occurs rapidly after drug exposure whereas total cell death requires longer incubation

Inventive Principle:
Principle #35Parameter changes

Solution Approach 2:

The patent replaces manual microscopy-based cell counting and viability assessment with automated flow cytometry analysis. This substitution enables rapid, high-throughput measurement of apoptosis markers in thousands of cells per second, reducing the time required to generate statistically significant results while maintaining accuracy

Inventive Principle:
Principle #28Mechanics substitution (Replace mechanical system)

2Productivity

If total cell death measurement is used, then drug effect is evaluated, but ability to distinguish tumor cell effect from normal cell effect is lost

Engineering Contradiction:
Improveevaluation speedVSAvoidcell type differentiation
Core Design Contradiction:
ProductivityVSMeasurement precision

Solution Approach 1:

The patent applies local quality by using cell-specific antibody markers (e.g., CD19 for B-cell lymphomas, CD34 for acute myeloid leukemia) to identify and separately analyze tumor cells versus normal cells within the same sample. This allows the measurement of apoptosis specifically in tumor cells exposed to drugs, providing precise differentiation of drug effects on malignant versus benign cells

Inventive Principle:
Principle #3Local quality

Solution Approach 2:

The patent introduces apoptosis-specific intermediaries (caspase antibodies, Annexin V) as mediators that bind selectively to apoptotic cells. These intermediaries serve as markers that can be detected by flow cytometry, enabling the distinction between drug-induced apoptosis in tumor cells versus normal cells based on the presence of these specific molecular markers

Inventive Principle:
Principle #24Intermediary (Mediator)

3Adaptability or versatility

If multiple drug combinations are evaluated, then treatment optimization is improved, but assay complexity and time requirements increase

Engineering Contradiction:
Improvedrug combination evaluationVSAvoidassay complexity
Core Design Contradiction:
Adaptability or versatilityVSDevice complexity

Solution Approach 1:

The patent segments the evaluation process into distinct modular components: (1) sample preparation with tumor cell enrichment, (2) parallel drug treatment of multiple aliquots, (3) flow cytometry measurement of apoptosis markers, and (4) automated data analysis. This segmentation allows multiple drug combinations to be evaluated in parallel across different sample aliquots, increasing versatility while managing complexity through standardized protocols

Inventive Principle:
Principle #1Segmentation

Solution Approach 2:

The patent creates a universal assay platform using flow cytometry that can measure apoptosis responses to any drug or drug combination by simply changing the drug treatment conditions. The same core methodology and instrumentation evaluate all drugs, providing multi-functionality that handles diverse drug combinations without proportionally increasing assay complexity

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

This approach provides faster turnaround times for cytotoxicity profiling, enabling more effective treatment choices and potentially increasing response rates, progression-free survival, and overall survival for cancer patients by quickly identifying optimal drug combinations and concentrations.

Implementation Method 1

measuring apoptosis or cell depletion in each of the at least 35 aliquots by flow cytometry

Methodology Applied
Scientific EffectFlow cytometry:

Data Source

PatentEP2433124B1Methods for providing personalized medicine tests ex vivo for hematological neoplasms
Publication Date: 2017.03.01 VIVIA BIOTECH SL
  • EP2433124B1 patent drawingFigure 1
  • EP2433124B1 patent drawingFigure 2
  • EP2433124B1 patent drawingFigure 3

AI summary

Described herein are methods, devices, and compositions for providing personalized medicine tests for hematological neoplasms. In some embodiments, the methods comprise measuring the efficacy of inducing apoptosis selectively in malignant cells using any number of potential alternative combination drug treatments. In some embodiments, the ex vivo testing is measured using a recently extracted patient hematological samples. In other embodiments, the efficacy is measured ex vivo using an automated flow cytometry platform. For example, by using an automated flow cytometry platform, the evaluation of hundreds, or even thousands of drugs and compositions, can be made ex vivo. Thus, alternative polytherapy treatments can be explored. Non- cytotoxic drugs surprisingly induce apoptosis selectively in malignant cells ex vivo. In some embodiments, the methods described herein comprise evaluating non-cytotoxic drugs.