Microfluidic CTC Detection via Hyperoxic Metabolic Labeling
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Current methods for detecting and quantifying circulating tumour cells (CTCs) and tumour cells (TCs) in liquid biopsies are inadequate due to low sensitivity and specificity, particularly for cells that have undergone epithelial-mesenchymal transition, and existing methods may generate false positives or negatives, and are not effective for early-stage cancer patients with low CTC burdens.
Innovation Solution
A method using a hyperoxic environment and a fluorophore-labelled metabolic indicator, specifically 2-NBDG, in combination with microfluidic chips for detection and quantification of CTCs/TCs, which exploits the metabolic differences between cancer and healthy cells based on fluorescence intensity.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Ease of operation
If EpCAM-targeting antibodies are used to capture CTCs, then detection is simplified, but CTCs with low or absent EpCAM expression are missed
Solution Approach 1:
The patent changes the detection parameter from targeting a specific antigen (EpCAM) to targeting a fundamental metabolic property (glucose uptake). By using a fluorophore-labelled glucose analog (2-NBDG) and creating a hyperoxic environment that enhances the Warburg effect, the method detects CTCs based on their hypermetabolic state rather than surface antigen expression, thereby capturing EpCAM-negative cells while maintaining operational simplicity through flow cytometry.
Solution Approach 2:
The patent introduces an intermediary substance (fluorophore-labelled 2-NBDG) that mediates the detection process. This glucose analog serves as a metabolic tracer that accumulates in hypermetabolic CTCs, allowing indirect detection of cancer cells through their metabolic activity rather than direct antigen-antibody binding, thus overcoming the limitation of EpCAM-negative cells.
2Reliability
If physical property-based methods (acoustic waves, microfluidics) are used to identify CTCs, then EpCAM-negative cells are detected, but specificity decreases generating false positives or negatives
Solution Approach 1:
The patent changes the detection parameter from physical properties (size, mass) to metabolic activity (glucose uptake rate). By exploiting the Warburg effect and enhancing it through hyperoxic conditions, the method achieves high specificity because normal blood cells do not exhibit the same hypermetabolic glucose uptake as CTCs, thereby reducing false positives while maintaining comprehensive detection.
Solution Approach 2:
The patent applies local quality by creating a hyperoxic microenvironment specifically in the detection chamber that enhances the metabolic difference between CTCs and normal cells. This localized oxygen enrichment amplifies the Warburg effect in CTCs, making their glucose uptake pattern distinct and highly specific to cancer cells rather than a general physical property.
3Reliability
If standard oxygen levels are used during incubation, then cell viability is maintained, but fluorescence intensity difference between cancer and healthy cells is insufficient
Solution Approach 1:
The patent changes the oxygen concentration parameter from standard atmospheric levels (21%) to hyperoxic levels ( significantly higher than 21% oxygen). This parameter change exploits and amplifies the Warburg effect, causing cancer cells to exhibit dramatically increased glucose uptake and lactate production under hyperoxic conditions, thereby generating sufficient fluorescence intensity difference for reliable detection while cells remain viable during the short incubation period.
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 allows for reliable and sensitive detection and quantification of CTCs/TCs, improving diagnostic accuracy and prognosis monitoring by distinguishing cancer cells from healthy cells through enhanced fluorescence intensity differences under optimized conditions.
Implementation Method 1
a fluorophore-labelled metabolic indicator (fluorophore-labelled 2-D-glucose derivative)... measuring the fluorescence of the fluorophore-stained cells
Implementation Method 2
incubating the cells contained in the liquid sample or the cells obtained in step a) with a solution supplemented with a fluorophore-labelled metabolic indicator and saturated with oxygen, during an appropriate time to allow the CTCs/TCs to accumulate the fluorophore-labelled metabolic indicator
Implementation Method 3
incubating the cells contained in the liquid sample or the cells obtained in step a) with a solution supplemented with a fluorophore-labelled metabolic indicator and saturated with oxygen
Data Source
Figure 1
Figure 2
Figure 3
AI summary
The invention relates to an apparatus and method of detecting and quantifying the number of circulating tumour cells (CTCs) and/or tumour cells (TCs) from a liquid biopsy by using a hyperoxic environment and incubation with a fluorophore-labelled metabolic indicator (fluorophore-labelled 2-D-glucose derivative) and microfluidic chips.